EURON and THEME joint PhD meeting

EURON and THEME 

joint PhD meeting 

University of Bonn 

September 

22 + 23 

2011

★ Euron local coordinators ★ 

Full partners 

The Netherlands 

• Maastricht University 

Dr. Jos Prickaerts 

jos.prickaerts@maastrichtuniversity.nl 

Germany 

• RWTH Aachen University 

Dr. Gary Brook 

gbrook@ukaachen.de 

• University of Bonn 

Prof. Dr. Jochen Walter 

jochen.walter@ukb.uni.bonn.de 

• Universität Köln 

Prof. Dr. Hannsjörg Schröder 

schroeder.anatomie@uni-koeln.de 

• Saarland University, Homburg 

Prof. Dr. Tobias Hartmann 

tobias.hartmann@uniklinikum-saarland.de 

Belgium 

• Katholieke Universiteit Leuven 

Dr. Tilmann Achsel 

tilmann.achsel@cme.vib-kuleuven.be 

• Université Libre de Bruxelles 

Prof. Dr. Roland Pochet 

rpochet@ulb.ac.be 

• Université catholique de Louvain 

Prof. Dr. Pascal Kienlen-Campard 

pascal.kienlen-campard@uclouvain.be 

• Universite of Liège 

Prof. Dr. Pierre Leprince 

pleprince@ulg.ac.be 

• Hasselt University 

Prof. Dr. Jean-Michel Rigo 

jeanmichel.rigo@uhasselt.be 

Associated partners 

• University of Leipzig, Germany 

Prof. Dr. Andreas Reichenbach 

reia@medizin.uni-leipzig.de 

• University of Minho, Portugal 

Prof. Dr. Joana Palha 

japalha@ecsaude.uminho.pt 

• CPN INSERM 

University Paris Descartes, France 

Dr. Laurence Lanfumey 

laurence.lanfumey@upmc.fr 

• Ege University, Izmir, Turkey 

Prof. Dr. Canan Y. Salam 

canan.y.salam@ege.edu.tr 

• Université Lille 1-Sciences et Technologies 

Prof. Dr. Michel Salzet 

m.salzet@orang.fr 

• Ondokuz Mayis University, Samsun, Turkey 

Prof. Dr. Suleyman Kaplan 

skaplan@omu.edu.tr

The local organizing committee: 

Prof. Dr. Jochen Walter 

Dr. Andrea Weber 

Prof. Dr. Volkmar Gieselmann 

Iris Ullrich 

Euron: 

Prof. Dr. Harry Steinbusch 

Dr. Nicole Senden 

Peggy Bisschoff 

Marie-Thérèse Moers 

EURON and THEME 

joint PhD meeting 

University of Bonn, Germany 

September 22-23, 2011

Welcome of THEME 

Dear EURON and THEME graduate students, Dear Colleagues, 

2 

EURON and THEME joint meeting 2011 

Welcome to the second annual THEME symposium in Bad Honnef, this year 

proudly hosted together with EURON, the European Graduate School of 

Neurosciences. 

This annual symposium has been designed to give PhD-students the 

opportunity to present their exciting research results to an international 

audience as either an oral or poster presentation. The curriculum is 

complemented by cutting edge talks of senior scientists, who will give insights 

into the challenges of fundamental research and demonstrate how to address 

and solve the underlying questions. 

I am grateful and excited holding this years THEME meeting together with 

EURON, This is a unique chance for an extensive dialogue between young 

and senior scientists on an international level. I encourage you to be part of 

lively discussions in the oral presentation as well as for the poster sessions. 

Take the chance to discuss your results. The scientific dialogue is an important 

basis for new and stimulating ideas and this in turn is the basis for a successful 

symposium. 

I wish all of us a pleasant and rewarding meeting! 

Prof. Dr. med. Volkmar Gieselmann

Information about THEME 

3 


THEME International Graduate School of Theoretical and Experimental 

Medicine 

The Faculty of Medicine in collaboration with the Faculty of Mathematics 

and Natural Science and the University Hospital of Bonn provide an excellent 

infrastructure for life science oriented research. The scientific and educational 

framework of THEME is built by renowned groups covering the main research 

fields of the Faculty of Medicine focused on: 

• Neuroscience 

• Genetics and Epidemiology of Human Diseases 

• Diseases of the Cardiovascular System 

Research encompasses a broad range of topics from molecular mechanisms 

in the development and progression of diseases to prevention, diagnosis and 

treatment. It also includes patient oriented clinical research. 

Courses of the study programme of THEME include lectures, seminars and 

practical courses and are integrated into the research fields mentioned above. 

THEME communicates relevant theoretical and practical skills needed for a 

successful thesis and prepares for a career in medical and life science research. 

THEME does not rely on external funding which specifically supports a graduate 

school. Rather it provides a framework for Ph.D. students working within 

peer reviewed projects mostly funded by the German Research Foundation 

(DFG), the EU of the German Ministry of Science. Therefore it is a permanent 

institution within the Medical Faculty of the University of Bonn which aims 

to continuously improve the conditions for a succesful accomplishment of an 

excellent Ph.D thesis.

Welcome Address of the Euron Director 

4 


I would like you to welcome you to our fifteenth EURON PhD days organized by 

the University of Bonn in Bad Honnef. 

It is for me a memorable event since it is our second EURON meeting that will be 

organized in Bad Honnef. The first one was at the ocassion of our masterclass/ 

workshop on Drugs and the Brain. For our new members from the University 

of Saarland, Homburg, and the University of Lille, I would like to remind that 

EURON has started already fifteen years ago with the idea to bring together 

neuroscientists in the Euregio to generate a platform for scientific and educational 

exchange for Master as well as PhD students and to stimulate a continuous life 

long learning programme. We initially have started with an educational program 

for PhD students and we could integrate all expertise of the scientists of our 

participating institutes. The educational program and the scientific collaborations 

attract the attention of potential new Master, PhD students and post-docs. 

Over the last fifteen years we were able to stimulate our collaboration through 

funding from the EU by two Marie Curie Early Stage Training Sites under the 

FP5 and FP6 programmes. Last year we received additional funding from the 

European Erasmus Life Long learning Program to develop within EURON a two 

years Research Master program “European Master in Neuroscience”. This Master 

will attract Master Students from all over Europe which later can start a PhD 

program within our network. 

The original educational idea is still valid and is even stronger these days, since 

nowadays also the EU and FENS have started within the NENS organization a 

focus on training for PhD students. Our new intitiatives encompass two new 

projects for which we will apply in 2012 on joint Master and PhD degrees. In 

addition the exchange of master and PhD students between the various centres 

is strongly encouraged and will be discussed further. However, it is not our aim 

to bring all scientific interests together under one common theme, moreover it 

is not necessary. Each of the participating institutes have their specific expertise 

which we will use in our joint applications. Scientifically EURON is moving from 

systems neuroscience towards more translational neuroscience. 

During this joint meeting with THEME we will have 29 oral presentations and 

40 poster presentations of the PhD students of our Graduate schools. We are 

pleased to offer jointly this platform to present your data to colleagues within

5 


the Graduate Schools and more important we hope you will use the opportunity 

to start your own networking. 

EURON can provide each of you with a EURON certificate on top of your local 

degree. This extra certificate will give you a premium on top of your local PhD 

degree and therefore better possibilities on the later post-doc market. It makes 

you special and it shows your premium quality. Information on the requirements 

to obtain this EURON certificate can be found in this booklet and on our website. 

We encourage all students to go for 3 to 6 months abroad to another lab and 

add or continue your research project within EURON. We offer opportunities, 

however it is you to decide to make use of them to your advantage. I wish you 

two scientific rewarding and also socially exciting days in Bonn / Bad Honnef. 

Prof. Dr. Harry Steinbusch, Director of EURON

6 

EURON and THEME joint meeting 2011

EURON 

a short introduction 

7 


Information from EURON 

8 


EURON currently consists of 10 full partner universities in Belgium, Germany and 

the Netherlands and 6 associated university partners in France, Portugal, Turkey 

and Germany. 

The research school EURON was officially established and accredited in 2003 

and successfully re-accredited in 2010 by the Royal Netherlands Academy of 

Arts and Sciences (KNAW). Euron has received at July 2010 its official approval 

(with delight) of this follow up recognition. The Committee was praising the 

international orientation and the way at which EURON tries to link the research 

and educational collaboration between the partners. This recognition is again for 

a six years period. 

Furthermore EURON has a strong international position due to its recognition by 

the EU as Marie Curie Early Stage Training Site during the period 2002-2010. 

Our new initiatives are: 

• To develop a joint Research Master program in Neuroscience between the 

EURON partner universities supported by a Curriculum Development project 

of the Erasmus Lifelong Learning Program. Estimated start date: September 

2012-2013. This joint master program will result in a joint degree. For Euron 

the EMiN program is the best example to share knowledge and expertise 

between universities and countries. 

• To (re) apply for a FP7 People Marie-Curie Training Network program with 

our SYNERGY project “Interdisciplinary Training Network in New Therapeutic 

Targets for Neurodegeneration” (January 12th, 2012). Innovative treatments 

can only result from innovative research. Therefore EURON has developed a 

new initiative in which seven universities and six large companies will work 

together to provide insight into the neurodegenerative processes and disorders 

in order to increase the development of new therapeutic strategies to prevent 

or slow down neurodegenerative processes. The complementary network 

of industrial and academic partners will combine their industrial or scientific 

knowledge: expertise and resources with a particular focus on the exchange of 

molecular, anatomical, electrophysiological techniques in neuroscience. 

• The realizations of joint PhD projects between EURON partner universities. In 

2009 EURON formally realized a so-called “Rotationsstellen” (PhD projects) 

between Maastricht University and the Klinikum Aachen and these joint PhD 

projects will now be also established between Maastricht University and 

Hasselt University called “sandwich positions”.

9 


• To apply for an Erasmus Mundus joint Doctorates April 2012 with the theme: 

CNS repair. Coordinators Hasselt and Maastricht University. 

EURON invests in a scientifically stimulating and inspiring environment for 

Master, PhD students and Post docs to perform disease oriented research and 

to be part of our extensive educationa program that reflects the expertise of our 

universities. 

EURON aims: 

• To stimulate mobility of students and teachers 

• To share knowledge and expertise 

• To transfer knowledge 

• To aim for uniformity of PhD degrees 

• To agree to the Bologna and Lisbon process 

The EURON Certificate is the way EURON encourages amongst others the 

uniformity in PhD degrees. Furthermore it will provide the EURON PhD students 

an extra recognition in their curriculum and that will be of advantageous for their 

further career. 

To obtain this certificate is depending on the following requirements: 

• PhD thesis in English language 

• Obligation to follow courses and to obtain credit points (based on the ects 

system: in 2012 this will be implemented. 10 ects from the EURON course 

program according to the entire PhD) 

• One EURON member (not from the home university) should represent EURON 

on the dissertation review committee and/or at the dissertation defense. 

• Scientific exchange visit(s) to other EURON research groups for a period of at 

least 3 months, related to the PhD program or other foreign experience. 

• The thesis should consist of at least four papers or of papers with a total Impact 

Factor of at least 10 as first-author or joint first author. Adjusted weight factors 

for the various disciplines in neuroscience are considered 

• Admittance to the EURON PhD degree program is limited to individuals holding 

a MSc, MD or MA degree or equivalent.

10 


EURON courses and events Autumn 2011 – 2012 

November 21, 2011 3th MHeNS Translational Workshop on Pain 

School for Mental Health and Neuroscience 

(MHeNS)/ Maastricht University 

November 24, 2011 EURON Course Rodent Neuroanatomy 

University of Cologne 

January 26-27, 2012 Euron course on Stereology in Neuroscience. 

Maastricht University / Ondukuz Mayis University, 

Samsun / Ankara University 

February 6-9, 2012 MHeNS Course From Neuroanatomy to 

Psychopathology 

MHeNS / Maastricht University 

April 16-20, 2012 FENS-IBRO/EURON Workshop Drugs and the 

Brain: an update in Psychopharmacology 

Maastricht University / University of Minho 

Sponsored by the ECNP 

Sept. - Oct. 2012 16th EURON PhD Days 

Maastricht University 

(dates need to be defined) 

Oct. – Nov. 2012 Update on Alzheimer Research: Workshop 

for PhD-students 

New Orleans, USA, Preceding SFN 2012 

EURON / ISAO / AHAF 

Information and registration courses and events: www.euronschool.eu

11 


List of EURON partner universities that have organized the EURON PhD Days in 

the past. 

2000: Maastricht University 

2001: Katholieke Universiteit Leuven 

2002: University of Cologne 

2003: Université de Liège 

2004: Saarland University, Homburg 

2005: Université de Liège 

2006: Maastricht University 

2007: Université catholique de Louvain 

2008: RWTH Aachen 

2009: Radboud University Nijmegen 

2010: Hasselt University 

2011: University of Bonn

12 


Programme 

13 


Location: Seminaris Hotel Bad Honnef 

Wednesday September 21 st 

13:00 - 13:20 Registration 

13:20 - 13:30 Opening and Welcome 

14 


13:30 - 15:30 Session I – (Chair: W. Kunz) 

O. Braganza Illuminating recurrent circuits in the epileptic dentate gyrus: a 

population Ca 2+ imaging study 

J. Anschlag Tcfap2c target genes in mouse primordial germ cells 

J. Lodder Characterisation of two N-acetylaspartylglutamate synthetases 

R. Hardt Functions of fatty acid 2-hydroxylation in mammals 

D. Gerlach Intracerebroventricular enzyme infusion corrects central 

nervous system pathology and dysfunction in mouse model of 

metachromatic leukodystrophy 

K. Schulte Simultaneous activation of the presynaptic cannabinoid CB 1 

receptor attenuates the function of the presynaptic muscarine 

M 2 receptor and the δ opioid receptor 

15:30 - 16:00 Coffee/tea 

16:00 - 17:40 Session II – (Chair: V. Gieselmann) 

M. Trautmann Inhibition of WNT signaling impairs growth of synovial sarcoma 

cells 

T. Nguyen Presenilins regulate autophagic induction 

V. Peeva Multiple mitochondrial DNA deletions in human diseases 

A. Bock The extracellular allosteric area of a seven transmembrane 

receptor controls intracellular signal trafficking 

C. Kilgus Local gene targeting and cell positioning using magentic nanoparticles 

for the generation of biological cardiac pacemakers 

17.40 - 17.50 Final Discussion 

17.50 - 18.30 Walking Dinner/Aperitive

Thursday September 22 nd 

9:00 - 10:00 Registration and poster set-up 

15 


10:00 - 10:10 Opening and Welcome 

Prof. Dr. med. Volkmar Gieselmann and Prof. Dr. Jochen Walter 

10:10 - 10:15 Communications 


10:15 - 11:00 Key lecture Prof. Dr. Eckhard Mandelkow 

MPI Hamburg/German Center for Neurodegenerative Diseases 

Bonn 

Structural principles of Tau aggregation and Tau-dependent 

neurodegeneration 

(Chair: J. Walter) 

11:00 - 12:20 Session I - Neurodegeneration 

(Chair: P. Kienlen-Campard) 

R. Gentier Expression of aberrant ubiquitin B (UBB +1 ) in the brainstem of a 

transgenic mouse model with AD-associated phenotype 

F. Dennissen Mutant ubiquitin (UBB +1 ) associated with neurodegenerative 

disorders is hydrolysed by UCH-L3 

C. Mencarelli The role of Goodpasture antigen-binding protein (GPBP) in the 

cellular response against Aβ-induced toxicity 

S. Tosheva γ-Secretase dependent phagocytosis of Amyloid-beta (Aβ) in 

microglial cells 

12:20 - 13:30 Lunch 

13:30 - 14:15 Key lecture Prof. Dr. Frank Kirchhoff 

University of Saarland 

Mechanisms of neuron-glia interaction in vivo – what transgenic 

mouse models tell us 

(Chair: H. Steinbusch) 

➤

16 


14:15 – 15:35 Session II – Neurodegeneration 

(Chair: M. De Baets) 

A. Sierksma Persistent spatial memory improvement after phosphodiesterase 

type 4d inhibition in the APPswe/PS1dE9 mouse model of 

Alzheimer disease 

T. Vanmierlo Brain-derived neurotrophic factor (BDNF), a bridge between 

depression and Alzheimer’s disease 

D.H. Zeef Memory deficits in transgenice huntington's disease rats 

R. Nalavade Investigating the role of miRNAs in Spinocerebellar Ataxia type 

3 (SCA3) 

15:35 – 16:00 Coffee/tea 

16:00 – 17:00 Session III – Behaviour 

(Chair: H. Beck) 

I. Carvalho Melo Role of Penk gene in stress reactivity 

I. Rayen Maternal fluoxetine exposure, regardless of prenatal stress, 

affects physiological systems involved in sexual development of 

offspring 

M.E. Siwek The influence of spatial distortion during body perception: an 

event-related potential study 

17:00 – 18:20 Session IV – Neurocommunication 

(Chair: J-M. Rigo) 

S. Paßlick Does the proteoglycan NG2 influence neuron-NG2 cell synaptic 

signaling? 

M. Vaessen Aberrant modular organization of cerebral functional networks 

in cognitive impaired children with frontal lobe epilepsy 

L. Pothmann Effects of antiepileptic drugs on hippocampal inhibitory 

microcircuits in the epileptic hippocampus 

C. Albus Altered hippocampal network oscillations in ASA-deficient 

mice 

18.30 – 19.00 Aperitive 

19:00 Walking Dinner + Posters

Friday September 23 rd 

17 


9.00 - 9:40 Session V – Neurocommunication (continued) 

(Chair: C. Steinhäuser) 

H. Dannenberg Labelling and optogenetic manipulation of learning-related 

neuronal assemblies 

M. Pabst Optogenetic modulation of the septohippocampal pathway 

9:40 - 10.20 Session VI – Genetics 

(Chair: C. Steinhäuser) 

S. Heilmann Identification of HDAC9 on chromosome 7p21.1 as a new 

candidate gene for androgenetic alopecia 

S. Horpaopan Identification of new causative genes in patients with 

adenomatous polyposis by copy number variation analysis 

10.20 - 10:40 Coffee/tea 

10:40 - 12:20 Session VII – Neuroinflammation 

(Chair: M. Theis) 

A. Maheshwari Local overexpression of Interleukin -11 in the central nervous 

system prevents demyelination 

C.-H. Chang Characterization of Kir channel expression in Schwann cells 

of the sciatic nerve in a mouse model of metachromatic 

leukodystrophy 

S. Smolders Migration of microglia in the embryonic neocortex 

L. Bodea Investigating the Role of Microglial TREM2 Receptor under 

Normal and Pathological Conditions 

E. Vlassaks The effects of asphyctic preconditioning and perinatal asphyxia 

on inflammation 

12:20 - 13:15 Lunch 

➤

18 


13:15 - 14:55 Session VIII – Neurodevelopment 

(Chair: V. Gieselmann) 

L. Mürtz Identification of microRNAs regulating differentiation of human 

neural stem cells 

B. Roese-Koemer The impact of bifunctional microRNA-9/9* on the differentiation 

of human ES cell – derived neural stem cells 

A. Avila Glycine receptor activation influences cortex development 

G. Bodea Unraveling the migratory behavior of dopaminergic neuronal 

subpopulations in the ventral midbrain 

A. Kabanova Determining the role of Shh signaling in establishing midbrain 

dopaminergic neuron subclasses 

15.00 - 15:15 Awards / Closing remarks

Poster presentations 

19 


1. Sven Akkerman Methodological considerations on exploration and 

discrimination measures of object recognition 

2. Buket Basmanav Rare variants on the schizophrenia associated 

1q21.1 microdeletion region 

3. Jessica Becker Genome-wide Meta-analysis on Dyslexia 

4. René Besseling Tract specific morphological and parametric 

reproducibility 

5. Eva Bollen TrkB agonist 7,8-dihydroxyflavone improves 

memory in an object recognition task 

6. Verena Borm Biochemical and functional properties of Liprinsalpha 

7. Claudia Cornelissen Immunohistochemical characterization of glial 

cells in human temporal lobe epilepsy 

8. Kimberly Cox Neurogenomics in perinatal asphyxia and fetal 

preconditioning 

9. Kristina Dobrindt A human iPS cell-based model for autosomal 

dominant hereditary spastic paraplegia 

10. Marianne Eisenhardt Extra-hepatic accumulation of CXCR3(+) NK cells 

as a novel mechanism of dysregulated NK cell 

function may contribute to immunopathogenesis 

of chronic HCV infection 

11. Daniela Evers Reprogramming of induced pluripotent stem cells 

from human cord blood 

12. Stephanie Friedrichs Cardiomyocytes obtained from murine induced 

pluripotent stem cells with long QT syndrome 3 

recapitulate typical disease-specific features in 

vitro

20 


13. Mary Gazea The role of primary cilia in the development of 

dopaminergic neurons in the murine ventral 

midbrain 

14. Jose Gerardo Nava Development of an in vitro system for the 

assessment of axon growth promoting properties 

of bioengineered scaffolds 

15. Andreas Glässner Natural killer cells from HCV-infected patients are 

highly efficient in inducing apoptosis of activated 

primary human hepatic stellate cells 

16. Alejandro Gomez Repair mechanisms at the neuromuscular 

junction. The role of Dok7 

17. Michaela Granzow Role of the Renin-Angiotensin-System in fibrosis 

and cirrhosis with portal hypertension in rats 

18. Stephanie Griemsmann Morphological and functional analysis of 

astrocytes in the thalamus 

19. Caroline Hammels Susceptibility and resilience in the social defeat 

model 

20. Verena Herl Bidirectional expression of Lck-GCaMP3 and 

DsRed in NG2 cells as an approach for monitoring 

glial Ca 2+ - microdomains 

21. Katia Herz Monitoring of vascular development in the 

murine embryonic stem cell system using a flt-1/ 

eGFP construct 

22. Dorothee Hodde Development of a Nanofibre-Based Tissue 

Engineering Strategy to Promote Functional 

Repair Following Traumatic Nervous Tissue injury 

23. Ali Jahanshahi Motor cortex stimulation induced neurogenesis

21 


24. Anna-Lena Klauke Targeting neuropathic pain by endocannabinoid 

signaling 

25. Sabine Klein HSC directed inhibition of Rho-kinase reduces 

portal pressure without systemic effects 

26. Sylvie van der Kruijs Abnormal functional connectivity between areas 

involved in emotion and executive control in 

psychogenic non-epileptic seizures (PNES) 

27. Andreas Lindstrot Gene expression profile of laser-microdisected 

prostate cancer tissues 

28. Roopika Menon Determining the Protein Profile of Prostate Cancer 

samples harboring the ERG rearrangement using 

MALDI Imaging Mass Spectrometry 

29. Kim Neitzert The role of the C-C chemokine CCL17 in Alzheimer’s 

disease 

30. Neville-Andrew Niessen Effect of prenatal stress and developmental 

fluoxetine exposure on hippocampal glucorticoid 

receptors and coactivator GR 

31. Astrid Ooms Function of BAG-3 in Skeletal and Cardiac 

Muscles 

32. Annika Ottersbach Nanomagneto-assisted cell tracking in vitro 

33. Geke Overvliet Cortical abnormalities and language impairment 

in Rolandic epilepsy 

34. Lutz Priebe Association between copy number variants in 

16p11.2 and major depressive disorder in a 

German case-control sample 

35. Sarah Rieck The application of different magnetic 

nanoparticles optimizes lentiviral transduction 

and cell positioning of endothelial cells

22 


36. Karin Rohleder Activity-dependent expression of presynaptic SV2 

proteins in hippocampal synapses 

37. Anna Schueth Retrograde tracing of neurons in the rodents 

lateral bladder wall - An experimental study with 

fluorescent latex beads: mucosal layers of the 

bladder vs. small intestinal mucosa 

38. Jo Stevens Cloning and production of anti inflammatory 

antibodies against A-BETA 

39. Svenja Ternes Generation of conditional knock out mice for 

diacylglycerol lipase α and β 

40. Sarah Vosen Radially symmetric endothelial cell replacement 

and lentiviral targeting in vessels by the use of 

magnetic nanoparticles (MNPs)

EURON Lecture 

Frank Kirchhoff 

ADDRESS 

Saarland University 

Department of Molecular Physiology 

Institute of Physiology 

Building 58 

66421 - Homburg 

Germany 

Email: frank.kirchhoff@uks.eu 

Homepages: http://physiology.uni-saarland.de 

Phone: +49 (0) 6841 - 1626489 

Fax: +49 (0) 6841 - 1626496 

Birthdate: 1.11.1960 

POSITION 

Professor of Molecular Physiology 

23 


1. Education 

1986 University of Hannover Diploma (Biochemistry) 

1990 University of Heidelberg Dr. rer nat. 

2. Positions and employment 

1991-1994 University of Heidelberg - Institute of Neurobiology 

Postdoc with Helmut Kettenmann, Heidelberg, Germany 

1995-1999 Max-Delbrück-Centrum for Molecular Medicine, 

Cellular Neurosciences, Berlin, Germany 

Postdoc and project leader with Helmut Kettenmann, Berlin

24 


2000-2009 Max Planck Institute of Experimental Medicine, 

Department of Neurogenetics, Göttingen, Germany 

Research group leader “Glial Physiology and Imaging” 

2009 - Saarland University, Medical Faculty, Institute of 

Physiology 

Full Professor for Molecular Physiology 

3. Other experience and professional memberships 

Editorial board Glia, since 2009 

Selection committee Studienstiftung des Deutschen Volkes, 2001 - 

and mentor 

Member German Physiological Society, German Neuroscience 

Society, German Society of Biochemistry, Society for 

Neuroscience 

4. Honours 

1981-1986 Fellowship „Studienstiftung des Deutschen Volkes“ 

1987-1989 Fellowship “Boehringer Ingelheim Fonds” 

5. Fields of Specialization 

Molecular and cellular mechanisms of neuron-glia interactions, transgenic 

mouse models, in vivo-imaging

ABSTRACT 

25 


Mechanisms of neuron-glia interaction in vivo – what 

transgenic mouse models tell us 

Our research focuses on the molecular and cellular mechanisms of neuron-glia 

interaction in the central nervous system. We are pursuing two main research 

questions: 

How do glial transmitter receptors sense and modulate synaptic transmission? 

What is the impact for living organisms? How do glial cells respond to acute 

injuries within the central nervous system? 

For functional analysis we generated (and are still continuing to develop) 

transgenic mouse models with cell-type specific expression of various fluorescent 

proteins (FPs) and inducible gene deletion. We are applying a combination of 

biochemical and molecular biological methods together with imaging techniques 

such as two-photon laser-scanning microscopy (2P-LSM) oder CCD imaging.

26 


THEME Lecture 

Eckhard Mandelkow 

POSITION 

Director and Scientific Member, Max-Planck-Society 

27 


1.Education 

Year Institution Degree Field of Study 

1964-66 Technical University of Braunschweig Vordiplom Physics 

1966-67 Tulane University, New Orleans Physics 

1967-69 University of Hamburg 

1970-73 Max-Planck-Institute for Med. Research, 

Diplom Physics 

Heidelberg Ph.D. Biophysics 

2. Positions and Honors 

1974-76 Postdoctoral fellow, Brandeis University, Waltham MA, USA 

1976-85 Group leader, Max-Planck-Institute for Medical Research, 

Heidelberg. 

1986-2011 Director, Max-Planck-Unit for Structural Molecular Biology, Hamburg 

2011 - Principal Investigator, DZNE, Bonn (German Center for Neurodegenerative 

Diseases), . 

Professor, University of Hamburg 

Scientific Member, Max-Planck-Society 

2010 Metlife Award 

2011 Potamkin Award

28 


3. Research fields 

Structure and function of the cytoskeleton, with special emphasis on 

• Structure of microtubules, actin filaments, intemediate filaments. 

• Structure of microtubule-associated proteins, including Tau. 

• Structures of protein kinases regulating microtubules. 

• Structures of microtubule motor proteins. 

• Axonal transport mechanisms. 

• Cell models of Tau pathology and neurodegeneration. 

• Screening and development of Tau aggregation inhibitors for therapy. 

Publications 

Mandelkow, E., Mandelkow, E.-M., Hotani, H., Hess, B., Müller, S.C. (1989). 

Spatial patterns from oscillating microtubules. Science 246, 1291-1293. 

Wille,H., Drewes,G., Biernat,J., Mandelkow,E.-M., Mandelkow, E. (1992). 

Alzheimer-like paired helical filaments and antiparallel dimers formed from 

microtubule-associated protein tau in vitro. J. Cell Biol. 118, 573-584. 

Drewes, G., Ebneth, A., Preuss, U., Mandelkow, E.-M., Mandelkow, E. (1997). 

MARK - a novel family of protein kinases that phosphorylate microtubuleassociated 

proteins and trigger microtubule disruption. Cell 89, 297-308. 

Kozielski, F., Sack, S., Marx, A., Thormählen, M., Schönbrunn, E., Biou, V., 

Thompson, A., Mandelkow, E.-M., Mandelkow, E. (1997). 

The crystal structure of dimeric kinesin and implications for microtubuledependent 

motility. Cell 91, 985-994. 

von Bergen, M., Friedhoff, P., Biernat, J., Heberle, J., Mandelkow, E.-M., 

Mandelkow, E. (2000). 

Assembly of tau protein into Alzheimer paired helical filaments depends 

on a local sequence motif (306-VQIVYK-311) forming beta structure. Proc. Natl. 

Acad. Sci. USA 97, 5129-5134. 

Bulic, B., Pickhardt, M., Schmidt, B., Mandelkow, E.-M., Waldmann, H., 

Mandelkow, E. (2009). 

Development of Tau aggregation inhibitors for Alzheimer disease. Angew. Chemie 

Int. Ed. 48, 2-15.

ABSTRACT 

29 


Structural principles of Tau aggregation and Taudependent 

neurodegeneration 

Eckhard Mandelkow 

DZNE, German Center for Neurodegenerative Diseases, c/o CAESAR, 53175 Bonn, Germany 

Tau is an unusual protein from several points of view: (1) Neurodegenerative 

diseases: Changes in Tau are early markers of AD and other "tauopathies" (e.g. 

FTD, Pick disease, PSP etc), (2) Neuronal cell biology: Tau is a brain-specific 

microtubule-associated protein (MAP), mostly confined to neuronal axons and 

implicated in neuronal differentiation, (3) Protein structure: Tau is the prototype 

of a "natively unfolded" or "intrinsically unstructured" protein, which does not 

require a defined structure for biological activity (in contrast to most "textbook" 

proteins with defined secondary and tertiary structure). We are interested 

in defining the interactions of Tau and the structural basis of its abnormal 

behavior in neurodegeneration. Tau is best known as an axonal protein that 

serves to stabilize microtubules, the tracks for long-haul traffic of vesicles and 

organelles in axons. As such, Tau interacts with tubulin (the building blocks of 

microtubules), motor proteins (kinesin, dynein), and several protein kinases and 

phosphatases that regulate these interactions. Malfunction of Tau (e.g. after 

hyperphosphorylation) can affect the stability of microtubules, interfere with 

motor-driven transport, and promote the pathological aggregation of Tau after 

detachment from microtubules. As a natively unfolded protein, Tau is not suitable 

for high resolution crystallography, however, the structure can be approached 

by spectroscopies (CD, fluorescence, FTIR, FRET) and NMR (solution and solidstate, 

collaboration with C. Griesinger, M. Zweckstetter, MPI Göttingen). The 

spectroscopic studies have revealed that Tau has "hotspots" for aggregation due 

to their tendency to form β-structure, and that Tau in solution has a "paperclip" 

folding where the N- and C-termini interact with the repeat domain. Both 

properties can be modified by phosphorylation at several sites. Knowledge of the 

β-forming hotspots enables one to design Tau molecules which favor or inhibit 

aggregation (pro- and anti-aggregant Tau). These variants now form the basis 

for designing cell- and animal models of tauopathy, and for screening inhibitors 

of Tau aggregation and other therapeutic agents (collab. B. Bulic, CAESAR). - 

Supported by MPG, DFG, VW Fnd, BMBF (KNDD), Metlife Fnd.

30 


Abstracts 

31 


32 


33 

Euron PhD meeting 2010 

Methodological considerations on exploration and 

discrimination measures of object recognition 

S. Akkerman 1,* , A. Blokland 2 , O. Reneerkens 1 , N.P. van Goethem 1 , E. Bollen 1 , 

H.J.M. Gijselaers 1 , C.K.J. Lieben 1 , H.W.M. Steinbusch 1 , J. Prickaerts 1 

1 Faculty of Health, Medicine and Life Sciences, Department of Neuroscience, School for Mental Health 

and Neuroscience, European Graduate School of Neuroscience (EURON), Maastricht University, The 

Netherlands; 2 Faculty of Psychology and Neuroscience, Department of Neuropsychology and Psychopharmacology, 

European Graduate School of Neuroscience (EURON), Maastricht University,The Netherlands. 

The object recognition task (ORT) is a frequently used tool in neurobiological 

research. In a sample trial, objects are presented to the animal. After a delay 

interval, one of the sample objects is replaced by a novel object in the test trial. 

Rats that remember the sample object(s) will spend more time exploring the novel 

object, compared to the sample object(s), the exploration difference is regarded 

to be a measure of memory. In this study, 28 ORT experiments were pooled, 

containing 731 male Wistar rats. We investigated the relationship between 3 

commonly used measures of discrimination and how they were related to levels 

of exploration in the sample and test trial. In this context, the effects of training, 

trial duration, delay interval and the amnesic drugs MK-801 (0.125 mg/kg) and 

scopolamine (0.1 mg/kg) on exploration and discrimination measures were 

investigated. Finally, we addressed object bias and relativity of object novelty 

as possible factors interfering with exploration and discrimination performance. 

Our analysis showed that the ORT is sensitive to potential biases like stress and 

side0-effects of drugs. There was no indication of a relationship between the 

level of exploration in the sample trial and discrimination performance. On the 

other hand, exploration in the test trial was positively related to the absolute 

discrimination measure but not to the relative discrimination measures, making 

them more resistant to exploration biases. Interaction with objects in prior 

sessions (training) decreased discrimination during subsequent 24 h retention 

interval testing. Thus, discrimination appears to reflect a lesser degree of 

familiarity of the novel object relative to the more familiar sample object due to 

a more recent encounter with the latter, rather than true novelty per se. Taken 

together, our findings suggest the consideration of pre-experimental exposure to 

objects, habituation to treatment procedures, balancing of object presentation 

and the use of relative discrimination measures when using the ORT.

34 


Altered hippocampal network oscillations in ASAdeficient 

mice 

Christina Albus 1 , Matthias Eckhardt 2 , Volkmar Gieselmann 2 , Heinz Beck 1 , Thoralf 

Opitz 1 

1 Laboratory for Experimental Epileptology, Department of Epileptology, Bonn, Germany; 2 Institute of Biochemistry 

and Molecular Biology, Bonn, Germany. 

Arylsulfatase A (ASA) is a lysosomal enzyme catalyzing the degradation of 

sulfatides. Mice deficient in ASA accumulate sulfatides in glial cells, microglia, and 

neurons and show neuromotor deficits as well as mild behavioral disturbances. In 

addition, invasive EEG recordings have revealed a marked cortical hyperexcitability, 

with episodes of spontaneous epileptiform activity. The phenotypic abnormalities 

have so far been mainly ascribed to the progressive demyelination and axonal 

damage, but if sulfatide accumulation exerts direct effects on neuronal excitability 

is so far unknown. 

To address this question, we first examined excitability on the network level in 

the hippocampal slice preparation. In the hippocampal CA1 and CA3 subfields, 

spontaneous slow episodes of population activity were observed under conditions 

of slightly increased excitability, termed sharp waves (SPW), but their incidence 

was not different in ASA-deficient mice compared to littermate controls. However, 

we observed a significantly higher fraction of SPWs with superimposed high 

frequency oscillations in the CA3 subfield of ASA-deficient mice (termed sharpwave 

ripples, SWRs). To address the cellular basis for increased SWRs, we have 

used intracellular recordings to examine the functional properties of pyramidal 

neurons in both the CA1 and CA3 subfields. We could not identify any systematic 

changes in passive or active membrane properties in hippocampal principal cells 

when comparing ASA-deficient mice to littermate control animals. 

These results suggest that ASA deficiency causes a selectively increased propensity 

to generate high frequency network activity within the subfield. Given the lack 

of functional changes in principal neurons, we propose that changes in inhbitory 

micronetworks may mediate the increase in high-frequency synchronized activity 

in ASA-deficient mice. 

Supported by the BMBF (Project LEUKONET)

35 

Euron PhD meeting 2010 

Tcfap2c target genes in mouse primordial germ cells 

Jana Anschlag 1 , Dawid Eckert 1 , Hubert Schorle 1 

1 Department of Developmental Pathology, Institute of Pathology, Bonn University Medical School, Sigmund-Freud-Str. 

25, 53127 Bonn, Germany. 

The transcription factor Tcfap2c (AP-2γ) is a member of the activating protein 2 

(AP-2) family of transcription factors and consists of a transactivation domain at 

the amino terminus and a helix-span-helix motif at the carboxyl terminus. The 

central basic region is responsible for dimerization and together with the helixspan-helix 

motif for DNA binding. It is known that AP-2 proteins bind to G/C rich 

regions like the palindromic sequences 5´-GCCN 3/4 GGC-3´ or 5´-GCCN 3/4 GGG-3´. 

During mouse embryogenesis Tcfap2c is expressed in primordial germ cells 

(PGCs) from embryonic day (E) 7.25 until E 12.5. Lack of Tcfap2c leads to sterile 

animals in which PGCs are specified but lost around E 8.0. In these PGCs the 

expression of germ cell markers is down-regulated and genes indicating somatic 

differentiation are up-regulated. Tcfap2c has been demonstrated to be upregulated 

in carcinoma in situ (CIS/IGCNU) and seminoma in humans, raising the 

critical question regarding the biological function and potential target genes of 

Tcfap2c in germ cells and testicular germ cells tumors. 

To investigate the function of transcription factor Tcfap2c in the transcriptional 

network of PGCs we used an in vitro differentiation protocol to generate PGCs. 

Mouse embryonic stem (ES) cells harbouring a Stella-GFP transgene were 

differentiated by embryoid body (EB) aggregation. Upon differentiation in EB 

culture, PGCs were identified and sorted by Stella-GFP signal a germ cell specific 

marker. 

Using these in vitro generated PGCs we performed a ChIP-on-chip analysis to 

search for promoters recognized by Tcfap2c. Preliminary analysis suggests that 

Tcfap2c regulates and probably reinforces the genetic and epigenetic network 

that is utilized to setup PGC identity. This experiment will help to further 

understanding of Tcfap2c in germ cell biology as well as in germ cell tumors.

36 


Migratory interneurons express functional glycine 

receptors during early development of the cerebral 

cortex 

Ariel Avila 1,2 , Pía Vidal 1 , Laurent Nguyen 2 and Jean-Michel Rigo 1 

1 BIOMED Research Institute, Hasselt University, Agoralaan C, Diepenbeek; 2 Developmental Neurobiology 

Unit, Centre for Cellular and Molecular Neurobiology, University of Liege, C.H.U. Sart Tilman, Liège, 

Belgium. 

The strychnine-sensitive glycine receptor (GlyR) is a member of the ligand-gated 

ion channel superfamily. In the adult, the GlyR is known to mediate fast inhibitory 

neurotransmission in the spinal cord and in the brainstem. The GlyR has also 

been described in the embryonic cortex from embryonic day 19 (E19) where it 

could participate in developmental processes, but its presence at earlier stages 

has not been documented. Since other neurotransmitter systems, i.e. GABA 

and its receptors, are known to be present during corticogenesis, we wondered 

if this could also be the case for glycine and its GlyR. In this study, we analyze 

the presence and physiological relevance of GlyR in the early development of 

the cortex using in vitro and ex vivo cultures of slices, patch-clamp, two photon 

microscopy for time lapse and for calcium imaging, immunocytochemistry and 

western-blot. Electrophysiological experiments confirmed the presence of GlyR 

mediated currents in migrating interneurons during early stages of development 

(E13-E15). Using in vitro labeling of interneurons we have described the 

pharmacological properties of glycinergic currents present in interneurons born 

at E13. The concentration-response curve showed an EC50 of 69 ± 12 micro M for 

glycine. All these currents were fully blocked by strychnine with an IC50 of 0.10 

± 0.02 micro M. Picrotoxinin and picrotin also blocked these currents, but with 

different potency, remaining 20 % of the current when 10 micro M of picrotin was 

used. Similar glycinergic currents were also observed in ex vivo preparations from 

Dlx5/6-Cre EGFP transgenic animals, where it was clear that GlyR expressing cells 

are a subpopulation of migratory interneurons. Consequently, immunostainings 

directed against the alpha 2 subunit of GlyR showed that 29 ± 2 % of cortical 

migrating interneurons, which were mainly born in the medial ganglionic 

eminence (MGE) at E13, express GlyR. All this evidences shows that GlyR appears 

earlier than ever described during cortex development and it is composed, mainly, 

by alpha 2 homomeric channels. It also shows that GlyR is not homogenously 

expressed and it is only present in a subpopulation of migrating interneurons 

born at a defined space-temporal window during brain development. In search 

for the physiological function of GlyR, two photon time lapse analysis for cell 

migration and calcium imaging was performed on ex vivo slices. All these studies

37 


were complemented by gain and loss of function experiments. As it has been 

previously described for the GABA system, we show here that GlyR play a role 

acting on the migratory behavior of interneurons and its effects are linked to 

modulation of calcium dynamic and the activity of calcium downstream targets. 

More in detail molecular mechanisms were analyzed by western-blot.

38 


Tract specific morphological and parametric 

reproducibility 

RMH Besseling 1,2,3 , JFA Jansen 1,2 , GM Overvliet 1,2,3 , MJ Vaessen 1,2,3 , PAM 

Hofman 1,2,3 , AP Aldenkamp 1,2,3 , WH Backes 1,2 

1 Department of Radiology; 2 School for Mental Health and Neuroscience, Maastricht University Medical 

Center (MUMC+), Maastricht, the Netherlands; 3 Epilepsy Center Kempenhaeghe, Heeze, the Netherlands. 

Rationale 

Tractography based tract segmentations were used to study reproducibility 

characteristics of tract volume as well as diffusion tensor metrics at the tract 

level. In addition, the relatively new diffusion contrast tract density imaging (TDI) 

was studied at the tract specific level. 

Furthermore, the reproducibility of the shape of the tracts (tract morphology) 

was assessed, which included the reproducibility of complete tract segmentations 

(extended) compared to tract segmentation excluding the directly subcortical 

projections (proximal). 

Methods 

Diffusion weighted imaging (DWI) was performed twice in 9 healthy subjects 

(28±6 y) on an 3T Philips Achieva scanner using: 128 gradient directions, voxel 

size of 2x2x2 mm 3 , and b=1200 s/mm 2 . Standard space seed and inclusion ROIs 

were mapped non-linearly to native DWI space, and subsequently constrained 

spherical deconvolution (CSD) probabilistic tractography was performed. The 

selected tracts were the genu of the corpus callosum, the cingulum, the pyramidal 

tract, the optic radiation, and the arcuate fasciculus. Tractograms were converted 

to tract density maps and thresholded to create tract segmentations that were 

used as ROIs to investigate tract fractional anisotropy (FA), apparent diffusion 

coefficient (ADC), volume and TDI. In addition, tract segmentations were mapped 

back to standard space to calculate between-session morphological overlap 

using the Dice similarity coefficient (DSC). Reproducibility of FA, ADC, volume 

and TDI was quantified using the coefficient of variation (COV) and the interclass 

correlation coefficient (ICC). 

Results 

For all metrics, reproducibility differed strongly between tracts and between 

proximal and extended tract segmentations. For FA, the proximal reproducibility 

values were: COV=1.6-2.7%, ICC=0.65-0.94. For ADC, comparable values were 

found (COV=1.5-3.5%, ICC=0.66-0.92). For volume, proximal COV=6.1-22%, 

proximal ICC=0.64-0.96 (0.029 for the arcuate fasciculus). For TDI, proximal

39 


COV=7.7-27% and not in all tracts accurate ICC values could be estimated. 

Morphological overlap as expressed by DSC ranged from 0.70 (arcuate fasciculus, 

extended) to 0.92 (corpus callosum, proximal). For a data subset of 66 gradient 

directions, DSC values were reduced for each tract with approximately 5%. 

Conclusion 

Estimates of tract FA and ADC are reliable, based on favorable COV and ICC values. 

Tract volume has worse COV values but maintains high ICC values. Tract TDI is the 

least reliable contrast, due to high COV combined with low ICC. 

Furthermore, our approach results in very high between-session morphological 

correspondence (up to DSC=0.92). Reliability is reduced in extended tract 

segmentations compared to the proximal segmentations. Since both types 

of segmentations differ primarily in their extent of subcortical projections, a 

reduced sensitivity in detection of subcortical changes in tract morphology can 

be expected.

40 


Conformational strain may underlie biased agonism 

of dualsteric ligands at the M 2 receptor 

Bock, A. 1 , Müller, A. 2 , Holzgrabe, U. 3 , De Amici, M. 4 , Kostenis, E. 2 , Mohr, K. 1 

1 Pharmacology and Toxicology Section, Institute of Pharmacy, University of Bonn; 2 Molecular, Cellular, 

and Pharmacobiology Section, Institute of Pharmaceutical Biology, University of Bonn; 3 Institute of Pharmaceutical 

Chemistry, University of Würzburg; 4 Dipartimento di Scienze Farmaceutiche ‘Pietro Pratesi’, 

Università degli Studi di Milano, Milano, Italy. 

G protein-coupled receptors (GPCRs) are seven transmembrane (7TM)-spanning 

proteins representing the largest and most ubiquitously expressed type of cell 

surface receptors. Many 7TMRs contain at least one allosteric binding site which 

is topographically distinct from the orthosteric site recognized by the respective 

endogenous messenger compound. The muscarinic M 2 acetylcholine receptor is 

an excellent model to study allosteric/orthosteric interactions as the core region 

of the allosteric binding site is well characterized. 

Recently, bisquaternary allosteric/orthosteric hybrid compounds were designed 

consisting of an allosteric inverse agonist fragment linked via an aliphatic 

hexamethylene middle chain with an orthosteric high affinity agonist. In 

exclusively activating the Gi pathway these dualsteric compounds showed biased 

agonism at the M 2 receptor [1] . However, hybrids’ potency was considerably lower 

than that of the orthosteric building block alone, suggesting a suboptimal fit of 

the hybrids’ building blocks to their corresponding binding sites. Here we show 

that middle-chain elongation to octamethylene considerably increases potency 

for M 2 receptor-mediated G protein activation as measured by [ 35 S]GTPγS binding 

in membranes of hM 2 -CHO cells. In order to check whether middle chain length 

affects biased signaling we carried out real-time measurements of dynamic mass 

redistribution in hM 2 -CHO cells using the EPIC® system (Corning, New York). Our 

findings show that the elongated hybrids regain modest ability for G s activation. 

We conclude that a spatial misfit between a dualsteric ligand and its corresponding 

orthosteric/allosteric receptor sites might impose a conformational strain on the 

receptor protein that underlies biased agonism of hexamethylene-type dualsteric 

compounds. 

[1] Antony J et al. (2009). Dualsteric GPCR targeting: a novel route to binding and 

signaling pathway selectivity. FASEB J 23: 442-450 

Support by the DFG is gratefully acknowledged (HO 1368/12-1, MO 821/2-1). 

This abstract is published in Naunyn Schmiedebergs Arch Pharmacol. 2011, 

March (383).

41 


Unraveling the migratory behavior of dopaminergic 

neuronal subpopulations in the murine ventral 

midbrain 

Gabriela O. Bodea and Sandra Blaess 

Institute of Reconstructive Neurobiology, Neurodevelopmental Genetics Group, University of Bonn, Sigmund-Freud-Str. 

25, D-53127 Bonn. 

Midbrain dopaminergic (MbDA) neurons located in the ventral tegmental area 

(VTA) and the substantia nigra (SN) are involved in many brain functions including 

reward associated behavior, modulation of emotions and motor control. MbDA 

neurons are derived from a Sonic Hedgehog (Shh)-expressing precursor domain 

in the ventral-medial mesencephalon. During development, they have to migrate 

from this precursor domain to form the laterally positioned SN and the medially 

located VTA. However, it is not known which migration patterns result in the 

formation of the distinct MbDA nuclei and which molecular mechanisms direct 

the migration of MbDA neurons. 

Using genetic inducible fate mapping to heritably mark Shh-expressing MbDA 

precursors and their descendants with YFP fluorescent reporter protein, we found 

that the ventral MbDA precursor domain can be roughly subdivided into a SN 

and VTA precursor domain. By following the fate of MbDA neurons derived from 

either the SN or VTA precursor domain during their migration phase (embryonic 

day 11.5-16.5), we are investigating whether the different MbDA subpopulations 

undergo distinct migration patterns to reach their final position in the SN or VTA. 

Our analysis of the position and morphology of fate-mapped MbDA neurons at 

different developmental time points indicates that SN neurons migrate radially 

while leaving their precursor domain and then switch to tangential migration 

to reach their final lateral position. In contrast, VTA neurons appear to migrate 

only radially. To monitor MbDA neuronal migration directly, we established an 

organotypic slice culture system of the embryonic midbrain, in which we can 

track the migratory behavior of fluorescently labeled, fate-mapped MbDA neuron 

subsets using time-lapse imaging. Observation of SN and VTA MbDA neurons 

in these slice cultures confirms their different migratory behavior. We are now 

investigating which molecular mechanism regulate radial versus tangential 

migration of MbDA neurons.

42 


Investigating the Role of Microglial TREM2 Receptor 

under Normal and Pathological Conditions 

Bodea Liviu-Gabriel 1 , Linnartz Bettina 1 , Colonna Marco 2 , Neumann Harald 1 

1 Institute of Reconstructive Neurobiology, University Bonn, 53127 Bonn, Germany; 2 Washington University 

School of Medicine, Department of Pathology & Immunology, St. Louis, MO 63110, USA. 

The triggering receptor expressed on myeloid cells 2 (TREM2) is presented on the 

cell surface of microglia, the resident immune cell of the central nervous system 

(CNS). The signaling cascade of TREM2 relies on the DAP12 adaptor molecule and 

especially on its immunoreceptor tyrosine-based activation motif (ITAM). TREM2/ 

DAP12 was shown to be involved in the non-inflammatory clearance of apoptotic 

neuronal material under normal conditions. In humans, a disease caused by loss 

of function mutation of either TREM2 or DAP12 was identified, the Nasu-Hakola 

disease, which is characterized by neuroinflammation in the CNS. 

The present study provides insights on TREM2 function and the mechanism of 

Nasu-Hakola disease by using a TREM2 knock-out (KO) mouse model. 

The nigrostriatal system of aged TREM2 KO mice shows signs of mild 

neurodegeneration. The data also show a slightly increase in microglial Iba1immunoreactivity 

in different brain regions of the aged TREM2 KO mice (ventral 

midbrain, hypothalamus, cortex) compared with the WT animals. However, TNFα, 

iNOS and IL-1β cytokine gene transcript levels were unaffected. 

Preliminary data show that the time frame of neurodegeneration was successfully 

shortened in TREM2 KO mice by using a protocol based on systemic challenges 

of mice with inflammatory stimuli,. Thus, injecting lipopolysaccharides (LPS) 

intraperitoneally on four consecutive days in mice has lead to dopaminergic 

degeneration but not in the vehicle treated controls. 

Data show that TREM2 KO mice are more susceptible to degeneration compared 

to WT controls, thus TREM2 presents a neuroprotective role within the CNS.

43 


BDNF signaling in hippocampal learning and memory: 

The effects of 7,8-dihydroxyflavone on object memory 

E. Bollen 1 , J. De Vry 1 , T. Vanmierlo 1 , H.M.W. Steinbusch 1 , J. Prickaerts 1 

1 Dept of Psychiatry and Neuropsychology, Maastricht University, The Netherlands. 

Brain derived neurotrophic factor (BDNF) has emerged as an important regulator 

of synaptic plasticity in the central nervous system. Binding of BDNF to its main 

receptor TrkB activates several intracellular signaling cascades, including the 

PLCγ-Ca 2+ pathway, the Ras-mitogen-activated protein kinase (MAPK) pathway 

and the phosphatidylinositol 3-kinase (PI3K)-Akt pathways, all of which have 

been implicated in neuronal growth and/or plasticity. In hippocampal long-term 

potentiation (LTP), one of the most commonly studied forms of plasticity which 

is generally considered as the cellular correlate of memory formation, BDNF 

has been attributed a critical role. In addition, in previous studies we found that 

the cAMP- and cGMP-mediated intracellular signaling cascades are exerting 

their specific effects on object memory consolidation via BDNF. Therefore, we 

studied the role of TrkB signaling pathways in synaptic plasticity and memory 

formation by injecting rats with 7,8-dihydroxyflavone (7,8-DHF; 0.3, 1 and 3 

mg/kg), a recently identified TrkB agonist during different consolidation phases 

after learning in an object recognition paradigm. Our results show that 7,8-DHF 

improves memory when injected both during the early and the late phase of 

object memory consolidation at 1 and 3 mg/kg. The lowest dose (0.3 mg/kg) 

resulted in a significant memory improvement only when injected at the late 

consolidation phase, which may suggest a stronger involvement of BDNF signaling 

in late memory consolidation. Taken together, these results show the potential of 

7,8-DHF as a cognition enhancer.

44 


Illuminating recurrent circuits in the epileptic dentate 

gyrus: a population Ca 2+ imaging study 

Oliver Braganza, Tony Kelly, Heinz Beck 

Department of Epileptology and Life&Brain Center, University Bonn, Sigmund-Freud-Str. 25, 53105 Bonn, 

Germany. 

Neural activity is defined by the anatomical and functional connectivity of the 

underlying neuronal networks. In epilepsy the neural network displays a tendency 

to spontaneous hypersynchronization, a phenomenon which may well be brought 

about by changes in the underlying network topology. A hallmark symptom of 

patients with temporal lobe epilepsy, as well as animal models thereof, is mossy 

fiber sprouting which is supposed to lead to profound changes in the network 

structure of the dentate gyrus. Physiologically there is a clear anatomical 

segregation between the granule cell input zone and their projections, the mossy 

fibers, reflected in the observation that there is very little interconnectivity 

between granule cells. In fact, the absence of such recurrent connections within 

the dentate gyrus granule cell layer is one of its hallmark features. In epilepsy, 

the anatomical segregation of input and output zone is severely impaired, as 

mossy fibers grow into the dendritic zone and occasional granule cells with basal 

dendrites, located in the output zone appear. We suggest that granule cells with 

basal dendrites exhibit a particularly high interconnectivity within the dentate 

gyrus, and thus may function as so-called network hubs. Modeling studies have 

shown that the presence of hub cells with extremely high connectivity alter the 

dynamics of networks profoundly, and can create hyperexcitability. In order to 

address questions of network topology it is necessary to simultaneously sample 

the activity of a large population of neurons with cellular resolution. We describe 

a multibeam multiphoton Ca 2+ imaging approach that permits fast imaging from 

large numbers of neurons in adult brain slices. We have used this approach to 

begin to address changes in network connectivity in the epileptic dentate gyrus.

45 


Characterization of Kir channel expression in 

Schwann cells of the sciatic nerve in a mouse model of 

metachromatic leukodystrophy 

Cin-He Chang 1 , Lihua Wang-Eckhardt 2 , Matthias Eckhardt 2 , Gerald Seifert 1 , 

Volkmar Gieselmann 2 , Christian Steinhäuser 1 

1 2 Institute of Cellular Neurosciences and Institute of Biochemistry and Molecular Biology, Medical Faculty, 

University of Bonn, Germany 

Metachromatic leukodystrophy (MLD) is a lysosomal storage disorder caused by 

deficiency of arylsulfatase A (ASA) and sulfatide storage. ASA deficient mice are 

an animal model for MLD, which, however, show a relatively mild phenotype. An 

improved mouse model of MLD, showing increased sulfatide synthesis, has been 

generated that displays increased sulfatide storage in neural cells of the brain 

and peripheral nervous system. In the peripheral nerves of these mice, hypo- and 

demyelination was observed, leading to a slowed propagation of action potentials 

(Ramakrishnan et al., 2007). Analysis of transcript and protein expression revealed 

an upregulation of the inward rectifying K + channel subunit Kir4.1 which in the 

nervous system is expressed solely by glial cells. To investigate whether these 

changes are accompanied by alterations in Kir4.1 channel function, we applied 

the patch clamp-technique to Schwann cells freshly isolated from sciatic nerves 

of ASA-/- mice, ASA-/- mice overexpressing the sulfatide synthesizing cerebroside 

sulfotransferase in Schwann cells (TG/ASA-/-), and wildtype littermates (WT) 

(postnatal day 300 – 500). 

Schwann cells from WT and ASA-deficient mice displayed depolarized membrane 

potentials of about -45 mV and a high input resistance (about 400 ΜΩ). These 

findings did not imply significant expression of functional Kir channels. In line with 

this assumption, neither WT nor ASA-deficient mice displayed Ba2+ (100 µM)sensitive 

inward currents at negative membrane potentials. In conclusion, data 

obtained so far suggest that in these MLD models, upregulation of Kir4.1 mRNA 

and protein in peripheral nerves is not accompanied by increased expression of 

functional channels. 

In the second part of my Ph.D. thesis I’m investigating the functional impact of 

inducible astrocyte-specific ablation of GABA B receptors. Analyses in WT mice 

revealed that the GABA B R1 subunit is expressed in almost all astrocytes of the 

hippocampus, whereas pyramidal neurons of the same brain region express 

both GABA B R1 and GABA B R2 subunits. Recent studies demonstrated that 

GABA B receptor activation in astrocytes leads to increase in [Ca 2+ ]i, release of 

gliotransmitters and modulation of synaptic signalling, although the underlying

46 


mechanisms are not understood. Analysis of GABA B R1 knockout mice will help 

to better understand how Ca 2+ signaling in astrocytes influences information 

processing in the brain.

47 


Neurogenomics in perinatal asphyxia and fetal 

preconditioning 

KEM Cox, E Strackx, E Vlassaks, M Sparnaaij, L Zimmerman, JS Vles, AW Gavilanes. 

Dept of Psychiatry and Neuropsychology, Maastricht University, The Netherlands. 

Background 

Part of perinatal hypoxic-ischemic brain damage can be attenuated when it is 

preceded by fetal preconditioning. The mechanism behind this endogenous 

neuroprotective phenomenon has not been deciphered yet. It is suggested that 

genomic reprogramming could be the answer to this question. We performed 

50 micro array experiments to evaluate changes in cerebral gene expression 

after fetal preconditioning and perinatal asphyxia at multiple time points. In this 

abstract we describe some preliminary results. 

Objective 

We aim to analyze the whole genome gene expression at multiple time points 

in pre- and neonatal pups after fetal preconditioning (FA) and perinatal asphyxia 

(PA). This can give us insight into the mechanisms of endogenous neuroprotection 

after FA and the deleterious effects of PA. Unraveling these mechanisms is an 

important step towards possible new clinical strategies for asphyctic neonates. 

Design/Methods 

FA was induced on E17 by clamping the uterine circulation of the maternal rat for 

30 minutes. On P0 pups PA was induced by placing the uterine horns, including 

the fetuses, in a water bath for 19 minutes. Control (CCD) and FA pups were 

delivered by Caesarean section. These procedures generated four experimental 

groups: CCD, FA, PA, and FA+PA. Five male pups per group were sacrificed at the 

following time points: 96h after FA, 6h and 96h after birth. RNA was isolated from 

the left hemispheres and 50 micro array experiments were performed on the 

Affymetrix platform. Up- or down-regulation of gene-products was studied via 

Gene-Ontology terms with a computational method named Gene Set Enrichment 

Analysis (GSEA). 

Results 

With GSEA we found a considerable number of GO-terms that were significantly 

up- or down-regulated compared to CCD animals (see figure 1). Interestingly in 

the protected phenotype (FA+PA) no GO-terms were significantly different from 

the control phenotype 6 hours after birth. Four days after perinatal asphyxia we

48 


see that numerous GO terms are down-regulated in the PA group. These data 

will be further analyzed and visualized in pathways; results are expected early 

September 2011. 

Conclusions 

These data in rodent fetuses and neonates are the first to demonstrate global 

gene expression profiles in the brain after fetal preconditioning and perinatal 

asphyxia. From previous behavioral studies we know that adult FA+PA animals 

perform similar to adult CCD animals despite the fact that the FA+PA group 

experienced a perinatal asphyctic insult. Interestingly here we also found no 

difference between these groups at 6hours after birth.

49 


Labelling and optogenetic manipulation of learningrelated 

neuronal assemblies. 

Holger Dannenberg, Milan Pabst, Karen van Loo, Albert Becker, Susanne Schoch, 

Heinz Beck 


Germany. 

Learning is thought to correlate with an initial change in the dynamics of a 

specific assembly of neurons and a subsequent sustained change in the synaptic 

connections between them. The propensity of this particular assembly of neurons 

to be recruited into synchronized activity can then be viewed as a memory trace. 

However, identifying these neuronal assemblies has been challenging. 

Immediate early gene promoters (IEGs) could be good candidates to genetically 

label cell populations active in-vitro, and to subsequently drive expression of 

fluorescent markers or light-sensitive channels that allow specific millisecondscale 

modulation of cellular activity. We have first compared the hippocampal 

expression patterns of the IEGs Arc (also known as Arg3.1), Egr1 (also known as 

zif268) and c-fos after a spatial learning paradigm in rats. We found an increase 

in the number of cells in the hippocampus of trained rats and/or rats having 

explored a novel environment compared to a cage control group. The expression 

patterns showed a partial overlap with subregional differences between each of 

the IEGs. 

We first chose a core promoter from the Arc gene (termed synaptic-activityresponsive 

element, SARE) to drive expression of the light-activated inhibitory 

chloride-pump halorhodopsin (eNpHR3.0) fused to EYFP. We show that the 

adenoviral approach is suitable to express eNpHR3.0-EYFP in principal cells of the 

hippocampus and characterized the SARE activation pattern by comparing it with 

the expression pattern of Arc, Egr1, and c-fos. eNpHR3.0-EYFP expressing cells 

could be inhibited by photo-activation in acute hippocampal slices, confirming 

its functionality in vitro. These approaches may permit the light-based activation 

or inhibition of neurons participating in the neuronal engram, both in-vitro and 

in-vivo.

50 


Hydrolysis of the mutant ubiquitin (UBB+1) 

accumulating in neurodegenerative disorders by 

UCH-L3 

F.J.A. Dennissen 1 , N. Kholod 1 , D.J.H.P. Hermes 1 , N. Kemmerling 1 , H.W.M 

Steinbusch 1 , N.P. Dantuma 2 and F.W. van Leeuwen 1 

1 Department of Neuroscience, Faculty of Health, Medicine and Life Sciences (FHML), Maastricht University, 

Maastricht, the Netherlands; 2 Department of Cell and Molecular Biology (CMB), Karolinska Institutet, 

Stockholm, Sweden. 

The ubiqutitin proteasome system (UPS) is of paramount importance for protein 

quality control since misfolded proteins are usually degraded by the UPS. The 

mutant ubiquitin UBB +1 accumulates selectively in the hallmarks of tauopathies 

and polyglutamine diseases. The UBB +1 protein is generated due to a frameshift 

at the mRNA level resulting in a loss of the C-terminal glycine and an addition 

of 20 amino acids to the C-terminus of this protein (Van Leeuwen et al., Science 

279, 242-247,1998). As a result, UBB +1 cannot be used for ubiquitination and 

impairs the UPS when being ubiquitinated. Furthermore, ubiquitinated UBB +1 

is refractory to deubiquitination by isopeptidase T. Studies in yeast and human 

cells showed that expression of UBB +1 gives rise to an additional C-terminally 

truncated product that corresponds in size with ubiquitin (Verhoef et al.,FASEB J. 

23, 123-33, 2009). In the transgenic UBB +1 mouse impaired contextual behaviour 

is observed ( Fischer et al.,Neurobiol.of Aging, 30, 847-863 2009) 

In order to identify the peptidase(s) responsible for the C-terminal truncation of 

UBB +1 we performed a systematic screen with 175 yeast deletion strains. After 

identifying the responsible enzyme for C-terminal truncation in yeast we cloned 

the mouse and human homologues and co-expressed them with UBB +1 in HEK293 

cells. We determined the effect of the individual enzymes on truncation of UBB +1 

using SDS-PAGE as was done for yeast. 

For yeast, we found the deubiquitylation enzyme YUH1 to be responsible for 

hydrolysis of the C-terminal extension of UBB +1 . Human and mouse homologue 

of YUH1, UCH-L3, were also able to hydrolyse the C-terminus of UBB +1 . Other 

members of the family, UCH-L1, -L4, -L5 or the naturally occurring human CRA_f 

isoform of UCH-L3 could not induce any truncation. 

Human and mouse UCH-L3 are able to hydrolyse the C-terminal extension of 

UBB +1 . Hydrolysis of UBB +1 s C-terminal tail prevents detection with the antibodies 

specific for this extension. Consequently, we hypothesize that UBB +1 which is 

detected in post-mortem tissue may be detectable due to lack of truncation by 

C-terminal hydrolyses (e.g. UCH-L3).

51 


The role of primary cilia in the development of 

dopaminergic neurons in the murine ventral midbrain 

Mary Gazea 1 , Christian Gojak 2 , Julia Franzen 2 , Kerry L. Tucker 2 and Sandra 

Blaess 1 

1 Neurodevelopmental Genetics, Institute of Reconstructive Neurobiology, Life and Brain Center, University 

of Bonn, Sigmund-Freud-Str. 25, 53127 Bonn; 2 Interdisciplinary Center for Neurosciences and Department 

of Anatomy, University of Heidelberg, 69120 Heidelberg, Germany. 

Midbrain dopaminergic (DA) neurons develop from the ventral midbrain floor 

plate. Sonic Hedgehog (Shh) signaling, which is mediated by the Gli zinc finger 

transcription factors Gli2 and Gli3, is essential for DA progenitor induction. 

Studies in spinal cord and forebrain have demonstrated that Shh signaling occurs 

in primary cilia and that Gli2 and Gli3 are not functional in absence of primary 

cilia. To assess whether primary cilia are important for Shh signaling in ventral 

midbrain development, we analyzed DA neuron development in cobblestone (cbs) 

mutants. Cbs mutants have reduced levels of the intraflagellar transport protein 

88 (Ift88) resulting in defects in ciliary function and in Shh signaling. Analysis of 

the developing ventral midbrain showed that the number of DA neurons and 

progenitors was severely reduced compared to wild-type. Interestingly, this 

phenotype was less severe than in mutants with inactivated Shh signaling. To 

investigate whether the mild phenotype in cbs mutants is due to residual Ift88 

function, we generated conditional knock-out (cko) mice in which the Ift88 

allele was inactivated in the midbrain after E8.5 (about a day after the onset of 

Shh signaling) and compared them with mutants with a conditional deletion of 

Gli2 and Gli3 (Gli2/Gli3 cko). In Ift88 cko midbrain, primary cilia were reduced 

innumber and deformed and Shh signaling appeared to be abolished. Ift88 cko 

had a smaller DA progenitor domain and the number of DA neurons was reduced 

by more than 50%. DA neurons and progenitors were also reduced in Gli2/3 

cko embryos, but the reduction was more severe than in Ift88 cko mutants. In 

summary, our data show that Ift88 plays an important role in the induction of 

ventral midbrain DA neurons, likely by maintaining functional primary cilia and 

consequently normal levels of Shh signaling.

52 


Autonomic dysfunction in Alzheimer’s disease 

(AD) correlated with UBB +1 in brainstem nuclei of 

transgenic mice 

R. Gentier, D.A. Hopkins, H.W.M. Steinbusch, F. W. van Leeuwen 

Department of Neuroscience, Faculty of Health Medicine and Life Sciences, Maastricht University, Maastricht, 

The Netherlands. 

Ubiquitin B +1 (UBB +1 ) accumulation in the cellular hallmarks of AD most probably 

contributes to a dysfunctional ubiquitin-proteasome system (UPS) (e.g. Van 

Leeuwen et al. 1998). Transgenic (tg) mice expressing the aberrant UBB +1 (line 

3413) in neurons show not only a behavioral phenotype (Fischer et al, 2009) 

but also an altered breathing pattern and the respiratory response to hypoxia 

is affected. In the present study, immunohistochemistry for UBB +1 neurons was 

mapped in the lower brainstem of the transgenic mice to determine if UBB +1 was 

expressed in pathways associated with respiratory modulation. Wild type control 

mice and absorbed UBB +1 antiserum showed no labeling. In line 3413, a clear 

and specific UBB +1 expression was present in various brainstem nuclei which 

are involved in the regulation of respiratory function. UBB +1 was observed in 

neurons in the dorsal vagal complex (area postrema (AP), nucleus of the tractus 

solitarius (NTS), dorsal motor nucleus of the vagus nerve). Strongly positive 

UBB +1 -immunoreactive neurons were present in subnuclei of the parabrachial 

nucleus (PBN), especially in the external lateral (PBNel) and dorsal lateral (PBNdl) 

subnuclei which receive visceral inputs from the caudal NTS. The rodent PBNel is 

involved in respiratory function and in the response to hypoxia and hypercapnia 

challenges (Song et al, 2009), functions that are affected in line 3413. These 

findings are consistent with the hypothesis that high UBB +1 expression might have 

mild toxic effects that could interfere with the normal functioning of pathways. 

AD patients suffer quite often from swallowing problems which frequently leads 

to aspiration pneumonia and eventually to disproportional mortality caused 

by autonomic dysfunction (Humbert et al, 2010). This is of particular interest 

because the phases of swallowing are regulated by central pattern generators 

in the brainstem and one of them is the NTS (Lang et al, 2009). This indicates 

a possible link between the respiratory aberrations shown in the UBB +1 tg mice 

and the respiratory/swallowing problems in AD patients because the same 

anatomical regions appear to be affected (Rüb et al, 2009).

53 


Intracerebroventricular enzyme infusion corrects 

central nervous system pathology and dysfunction in 

a mouse model of metachromatic leukodystrophy 

Stijn Stroobants, Debora Gerlach, Frank Matthes, Dieter Hartmann, Jens Fogh, 

Volkmar Gieselmann, Rudi D’Hooge and Ulrich Matzner 

1 Laboratory of Biological Psychology, Department of Psychology, University of Leuven, Tiensestraat 102, 

B-3000 Leuven, Belgium; 2 Institut für Biochemie und Molekularbiologie, Rheinische Friedrich-Wilhelms- 

Universität, Nussallee 11, D-53315 Bonn, Germany; 3 Anatomisches Institut, Rheinische Friedrich-Wilhelms-Universität, 

Nussallee 10, D-53315 Bonn, Germany and 4 Zymenex A/S, Roskildevej 12C, DK-3400 

Hillerød, Denmark. 

Metachromatic leukodystrophy (MLD) is a lysosomal storage disease caused 

by a functional deficiency of arylsulfatase A (ASA). This leads to storage of the 

membrane lipid 3-O-sulfogalactosylceramide (sulfatide), which is abundant 

in myelin. The enzyme deficiency causes demyelination of the PNS and CNS, a 

progressive multisystemic pathology and premature death. ASA-deficient mice 

have been generated as an animal model of MLD. ASA knockout mice store sulfatide 

and develop progressive neurological symptoms, yet no demyelination. The mild 

disease phenotype could be aggravated and demyelination could be induced by 

a transgenic overexpression of the galactocerebroside sulfotransferase. Enzyme 

replacement therapy (ERT) is an option to treat MLD and other lysosomal storage 

diseases. In ERT, the nonfunctional enzyme is replaced by active recombinant 

enzyme using repeated and lifelong injections. This approach is, however, limited 

by the blood-brain barrier (BBB) which prevents efficient trespass of injected 

enzyme from the circulation to the brain parenchyma. To circumvent the BBB and 

improve delivery of therapeutic enzyme to the CNS we used osmotic miniature 

pumps and infused recombinant human ASA (rhASA) directly into the brains of 

conventional and genetically aggravated ASA knockout mice. rhASA continuously 

delivered to the lateral ventricle for 4 weeks penetrated the ependyma and brain 

parenchyma. Co-immunostaining of rhASA, cellular marker proteins and lamp-2 

indicated efficient endocytosis and lysosomal targeting of rhASA by all cell types 

of the Brain. Alcian blue staining of brain sections revealed complete reversal 

of storage in the infused hemisphere. rhASA concentrations and sulfatide 

clearance declined with increasing distance from the infusion site. Correction 

of the ataxic gait indicated reversal of central nervous system dysfunctions. 

The profound histopathological and functional improvements, the requirement 

of low enzyme doses and the absence of immunological side effects suggest 

intracerebroventricular ERT to be a promising treatment option for MLD and 

other LSDs with prevailing CNS disease.

54 


Repair mechanisms at the neuromuscular junction. 

The role of Dok7 

Alejandro Gomez, Jo Stevens, Peter Molenaar, Mario Losen, Marc H. De Baets, 

and Pilar Martinez-Martinez. 

Department of Neuroscience. Maastricht University. 

Myasthenia gravis (MG) is an autoimmune disease that is caused by antibodies 

against the acetylcholine receptor (AChR) at the neuromuscular junction (NMJ). 

The disease is caused by a continuous attack of the auto-antibodies in the 

postsynaptic membrane of the NMJ, where the AChR is located at high density. 

Antibody binding leads to damage of the synapse, to AChR loss and to loss of 

AChR-associated proteins. 

Expression of less active forms of AChR subunits or of the AChR-associated 

proteins rapsyn, MuSK and Dok7 are the cause of congenital myasthenic 

syndromes (CMS). Therefore the loss of these proteins in autoimmune MG could 

aggravate the disease. The muscle responds to an autoimmune attack by upregulating 

the proteins that can compensate for the loss of function (e.g. AChR 

subunits, Dok7). However, in MG these repair mechanisms might be insufficient 

and some patients have severe disease despite of low antibody levels. 

MuSK and its activator Dok7 are very important factors for the dense clustering 

of AChR at the postsynaptic membrane. To study the relevance of this clustering 

machinery in the context of MG, we will investigate both the silencing and overexpression 

of Dok7 in the animal model of MG (experimental autoimmune 

myasthenia gravis, EAMG). To this end, we designed siRNA’s that targets Dok7 

and tested them in vitro for their silencing efficiency. Afterwards, we will transfect 

the validated siRNAs into the tibialis anterior muscle of rats by electroporation. 

We hypothesize that silencing Dok7 expression in the adult muscle leads to an 

increased susceptibility to passive-transfer EAMG and a delayed recovery from the 

EAMG. Besides, we will study if Dok7 over-expression in the muscle increases the 

resistance to EAMG. We will evaluate the animal’s neuromuscular transmission 

by electromyography, quantify AChR content at the NMJ by radioimmunoassay, 

and study the morphology of the endplate by electron microscopy.

55 


Epigenetic marks in brains of susceptible and resilient 

mice after social defeat stress 

Caroline Hammels 1,2 , Jos Prickaerts 2,3 , Tim Vanmierlo 2,3 , Gunter Kenis 1,2 , Harry W. 

M. Steinbusch 2,3 , Jim van Os 1,2,4 , Daniel van den Hove 2,3 , Bart Rutten 1,2 

1 Dept. of Psychiatry and Psychology, School for Mental Health and Neuroscience, Maastricht University, 

Maastricht, The Netherlands; 2 European Graduate School of Neuroscience (EURON), Maastricht University, 

Maastricht, The Netherlands; 3 Dept. of Neuroscience, School for Mental Health and Neuroscience, 

Maastricht University, Maastricht, The Netherlands; 4 King’s College London, Institute of Psychiatry, London, 

United Kingdom. 

When subjected to aversive life events, most individuals do not develop 

depressive-like psychopathology. The molecular mechanisms underlying this 

resilience remain to be elucidated. Yet, recent studies using social defeat as a 

mouse model for depression suggest that epigenetic mechanisms might be 

involved. We used the chronic social defeat paradigm in which eight-week-old 

experimental C57Bl6 are exposed for 10 consecutive days to an aggressor CD1 

mouse for ten minutes daily, followed by a 24 hours sensory (but not physical) 

contact. Social avoidance was measured by the social interaction test. We found 

that 50% of the mice showed a susceptible phenotype, characterized by social 

avoidance, while the other 50% showed a resilient phenotype in this test, which 

is in line with previous studies. Defeated mice did not display anhedonia, as 

measured by the sucrose intake test or increased anxiety-like behavior in the 

elevated zero maze. On the other hand they do display increased despair in 

the forced swim test. These data support previous findings displaying a prodepressant 

effect of the social defeat paradigm. However, effects of social defeat 

are generally reported on anhedonia instead of despair. Furthermore, repeated 

social avoidance testing revealed that ‘resilience’ can change over time, i.e. some 

resilient mice can become susceptible and vice versa. This interesting observation 

needs further research, in particular on the question which selection criterium is 

most valid. In order to start determining whether certain epigenetic mechanisms 

are associated with (and possibly mediate) these behavioral differences after 

social defeat, immunohistochemical stainings for epigenetic marks are currently 

performed.

56 


Functions of fatty acid 2-hydroxylation in mammals 

Robert Hardt, Volkmar Gieselmann, Matthias Eckhardt 

Institut für Biochemie und Molekularbiologie (IBMB), Uniklinikum Bonn, Nussallee 11, 53119 Bonn, 

Germany. 

Sphingolipids represent a highly diverse group of essential eukaryotic membrane 

lipids. They vary mainly in their type of headgroup, sphingoid base or acyl chain 

(i.e. length, saturation status, hydroxylation). 

In our group we focus on the role of sphingolipids containing N-linked 

α-hydroxylated fatty acids (hFA). In mammals there has been described only 

one enzyme until now, fatty acid 2-hydroxylase (FA2H), responsible for fatty acid 

α-hydroxylation. FA2H is expressed in various tissues mainly brain, spinal cord, 

skin, testis, ovary, kidney, stomach and intestine corresponding with significant 

amounts of hFA-sphingolipids. Contrasting their high incidence and diverse 

distribution, their functional roles in the organism are still largely unknown. 

One aspect of interest is the possible influence of α-hydroxylation on transport, 

metabolic processing and membrane distribution of sphingolipids within the 

cell. For example there are experimental observations indicating that GlcCer 

and hFA-GlcCer differ substantially in these aspects. For this reason sphingolipid 

α-hydroxylation may represent a key mechanism for generating membranes of 

differential composition. This in turn will affect a membrane’s physical properties 

and probably the distribution of membrane and membrane-associated proteins. 

Therefore using two previously described lipid transport assays (Halter et al. 

2007) the possible differences in GlcCer and hFA-GlcCer transport are examined 

in detail. 

In addition studies on FA2H knock-out mice, created in our group, showed that 

some hFA-sphingolipids are still present in certain tissues (esp. skin), implying 

the existence of at least one additional fatty acid 2-hydroxylase. To discover the 

second enzyme a complementation assay utilizing a mouse skin cDNA library and 

FA2H-deficient S. cerevesiae strains is established. 

References: 

Halter et al. 2007. Pre- and post-Golgi translocation of glucosylceramide in 

glycosphingolipid synthesis. Journal of Cell Biology 107:101-115

57 


Identification of HDAC9 on chromosome 7p21.1 as a 

new candidate gene for androgenetic alopecia 

Stefanie Heilmann 1,2* , Felix F. Brockschmidt 1,2* , Justine A. Ellis 3,4 , Sibylle 

Eigelshoven 5 , Sandra Hanneken 5 , Christine Herold 6 , Susanne Moebus 7 , 

Margrieta A. Alblas 1,2 , Bärbel Lippke 1,2 , Nadine Kluck 1,2 , Lutz Priebe 1,2 , Franziska 

A. Degenhardt 1,2 , Rami Abou Jamra 8 , Christian Meesters 6,9 , Karl-Heinz Jöckel 7 , 

Raimund Erbel 10 , Stephen Harrap 3 , Johannes Schumacher 2 , Holger Fröhlich 11 , 

Roland Kruse 12 , Axel M. Hillmer 13 , Tim Becker 6,9 , Markus M. Nöthen 1,2 

*these authors contributed equally to this work 

Institute of Human Genetics, Biomedical Center, University of Bonn, Germany. 

Androgenetic alopecia (AGA) is the most common form of hair loss in humans 

affecting up to 80% of European males by the age of 80 years. Candidate gene 

analyses and genome-wide association studies (GWAS) have identified two major 

genetic risk loci: the X-chromosomal androgen receptor (AR)/ectodysplasin A2 

receptor (EDA2R) locus and the paired box 1 (PAX1)/forkhead box A2 (FOXA2) 

locus on chromosome 20. Although these loci explain a significant fraction 

of the overall genetic risk for AGA, additional genetic risk factors still await 

identification. Here, we performed a GWAS using an case-control sample of 

581 severely affected AGA cases and 617 controls, 270 of whom were men 

aged >60 years with no signs of AGA, representing the 20% least affecteds in 

the population. The best association signal was obtained for rs756853, located 

intronically in the histone deacetylase 9 (HDAC9) gene on chromosome 7p21.1. 

Fine mapping analysis within the case-control sample and a corresponding familybased 

analysis revealed rs756853 and rs2249817, respectively as the most highly 

associated variants. The association finding for rs2249817, located 6kb distally of 

rs756853, was confirmed within an independent Australian sample (P=0.026). A 

combined analysis of severely affected German and Australian cases (N=639) and 

unaffected controls (N=384) for rs2249817 revealed a strong association signal of 

P=9.09x10 -8 , odds ratio 1.63 [1.36-1.95]. Tissue expression studies demonstrated 

HDAC9 expression in AGA related tissues like scalp and hair follicle. Genotypespecific 

expression as well as splice studies revealed no strong genotypic effects, 

although smaller effects cannot be excluded. Pathway analyses however support 

the hypothesis that HDAC9 plays a functional role in AGA via interaction with the 

AR gene. The genetic data of the present study thus provide strong evidence that 

HDAC9 is the third AGA susceptibility gene.

58 


Development of a Nanofibre-Based Tissue Engineering 

Strategy to Promote Functional Repair Following 

Traumatic Nervous Tissue Injury 

Hodde D 1 , Kriebel A 2 , Mey J 2,3 , Klee D 4 ; Möller M 4 ; Weis J 1 , Brook GA 1 . 

1 Institute of Neuropathology, RWTH Aachen University Hospital, Germany; 2 Institute of Biologie II, RWTH 

Aachen, Germany; 3 National Hospital of Paraplegia, SESCAM, Toledo, Spain; 4 DWI e.V. and Institute of 

Technical and Macromolecular Chemistry, RWTH Aachen, Germany. 

Peripheral nerve injury (PNI) causes an immediate loss of function. Simple 

transection injuries can be surgically repaired with tensionless sutures. However, 

the presence of a larger gap within the lesioned nerve requires the interposition 

of a bridging substrate or conduit. The current “gold standard” treatment for the 

repair of such gaps in lesioned peripheral nerve is the autologous nerve graft 

which has significant limitations. Therefore, alternative strategies to replace or 

at least supplement the autograft are desired. The goal of the present project is 

to develop a biomimetic nanofibre-based implantable scaffold that will induce 

Schwann cell migration and axonal regeneration to promote functional repair 

after PNI. Using the electrospinning technique we have generated simple two 

dimensional (2D) arrays of highly aligned poly-caprolactone (PCL) nanofibres that 

allow the investigation of single cell-single fibre interactions in vitro. By adapting 

the recently published methodology of Yang et al. (2011), the 2D arrays of highly 

aligned nanofibres have been collected and stacked onto each other resulting in 

a three dimensional (3D) configuration of layered nanofibres that are suspended 

in air. These 3D arrays appear to be remarkably stable, even when infused with 

a number of different hydrogels (i.e. gelatine, fibrin and the Puramatrix self 

assembling nanofibre hydrogel). Infusion with these hydrogels resulted in no 

disruption of the layered architecture, nor of the orientation of the nanofibres. 

Cell-substrate interactions in simple 2D in vitro investigations have demonstrated 

the powerful orientating influence of such nanofibres on Schwann cell growth, 

process formation and length of extension. Ongoing investigations are focussing 

on Schwann cell-nanofibre interactions in the present 3D arrays. Such nanofibrebased 

devices represent a significant advance in the field of tissue engineering 

and, if demonstrated to be effective in controlling and directing glial and axonal 

growth in vitro, will be used for future implantation experiments as nerve bridges 

in vivo.

59 


Copy number variation analysis in 134 unrelated 

patients with mutation negative adenomatous 

polyposis 

Sukanya Horpaopan 1 , Stefanie Vogt 1 , Isabel Spier 1 , Alexander M Zink 1 , Kirsten 

Wöllner 1 , Stefan Herms 1,2 , Markus Draaken 1,2 , Sandra Pasternack 1 , Markus M 

Nöthen 1,2 , Per Hoffmann 1,2 , Stefan Aretz 1 

1 2 Institute of Human Genetics, University of Bonn, Germany; Dept. of Genomics, Life & Brain Center, 

University of Bonn, Germany. 

Background 

Adenomatous polyposis syndromes are characterised by multiple colorectal 

adenomas and a high lifetime risk of colorectal cancer. In up to 50% of patients 

with colorectal adenomatous polyposis no germline mutation in the currently 

known genes – APC and MUTYH – can be identified. Copy number variants 

(CNVs) have recently been recognised as important forms of structural variation 

which also predispose to human disease. It can be hypothesised that in 

particular heterozygous microdeletions contribute to the underlying cause in yet 

unidentified genes responsible for adenomatous polyposis syndromes. 

Methods 

Genomic DNA from 134 unrelated mutation negative polyposis patients was 

used for genome-wide SNP genotyping with the HumanOmni1-Quad BeadArray 

(Illumina). Putative CNVs were identified by the QuantiSNP v2.2 algorithm, filtered 

according to various criteria by use of the Cartagenia Benchsoftware, by in-silicoanalysis, 

and by comparison with 531 healthy controls, and validated by qPCR. 

Results 

35 unique heterozygous deletion CNVs containing 38 protein coding genes could 

be validated in 33 patients (25%) but not in healthy controls. 25 genes are partly 

or completely deleted, in 13 more the deletion affects intronic regions only. 

Additionally, 47 unique duplication CNVs from 38 patients (28%) were validated by 

qPCR. 49 out of the 106 involved genes are partially duplicated which might point 

to potential loss-of-function effects. All CNVs are present only once in the whole 

cohort; all except eight patients harbor just one CNV. Candidate adenoma genes 

include protein kinases, transcription factors, and potential tumor suppressors. 

Conclusions 

By applying stringent filter criteria, we identified a group of rare deletion 

and duplication CNVs which might contain predisposing genes for adenoma

60 


formation. After prioritization of the included genes according to expression 

profiles, function, and pathway, currently work includes sequencing the coding 

regions of the most interesting candidates in all patients to look for pathogenic 

point mutations. The study was supported by the German Cancer Aid (Deutsche 

Krebshilfe).

61 


Hyperdopaminergic status in Huntington’s disease 

Ali Jahanshahi 1,2,3 , Rinske Vlamings 1,2,3 , Dagmar Zeef 1,2,3 , Harry Steinbusch 1,3 and 

Yasin Temel 1,2,3 

Departments of 1 Neuroscience and 2 Neurosurgery, Maastricht University Medical Centre, Maastricht, 

The Netherlands; 3 European Graduate School of Neuroscience (EURON). 

Introduction 

Huntington’s disease (HD) is a neurodegenerative disorder characterized by 

progressive cognitive impairments and chorea. The latter has been linked to 

an increased dopaminergic neurotransmission in the striatum. Treatment with 

dopamine (DA) antagonist or DA depleting drugs can reduce chorea. However, 

the origin of this hyperdopaminergic status remains unknown. Tracing studies 

have shown that dopaminergic input to the striatum comes from the substantia 

nigra pars compacta (SNc), ventral tegmental area (VTA), and a specific cell 

population of the dorsal raphe nucleus (DRN). We tested the hypothesis that 

elevated striatal DA levels are caused by changes in the number of dopaminergic 

neurons in the SNc, VTA and DRN in a transgenic rat model of HD (tgHD) and in 

the DRN of human HD specimens. 

Materials and methods 

Rodents were transgenic HD (tgHD) rats (homozygous, hemiozygous HD 

and wildtype littermates). Brains were cut serially and processed for 

immunohistochemical staining. Sections containing the striatum, VTA, SNc, and 

DRN were immunohistochemically processed for tyrosine hydroxylase (TH), the 

rate-limiting enzyme in the synthesis of DA. In addition, we processed another 

series of brain sections containing the DRN for phenylalanine hydroxylase (PH8) 

immunohistochemistry, rate limiting enzyme in serotonin production. The number 

of TH-ir and PH8-ir cells in tgHD rats was evaluated using Stereology and the level 

of TH expression in the striatum were assessed by optical densitometry. 

Human brain sections of HD patients and controls containing the DRN were 

stained for TH and PH8. The number TH and PH8 containing cells in the DRN 

were evaluated semi-quantitatively. 

Results 

The number of TH-ir cells in the tgHD rats was significantly higher as compared 

to the WT littermates, in all investigated regions in tgHD rats. Measurements of 

the optical densities showed that the TH-ir in the dorsal and ventral striatum 

was significantly higher in the tgHD rats as compared to the WT counterparts.

62 


In addition, our results showed substantial decrease in the number of PH8-ir 

neurons in the DRN. The number of TH-ir cells in the DRN of human HD brains 

was significantly higher compared to the human controls and was accompanied 

by a reduction in the number of PH8-ir cells. 

Conclusion 

We found that the origin of increased levels of dopamine in the striatum might 

be linked to an increase in the number of TH-ir cells in the VTA, SNc and the DRN 

of tgHD rats. In addition, we observed increased number of TH-ir and reduced 

number of PH8-ir cells in the DRN of tgHD rats and HD patients. We suggest that 

the underling mechanism for this hyperdopaminergic status in HD can be due to 

a change in phenotype of the non-dopaminergic cells, like serotonergic cells into 

dopaminergic cells.

63 


Determining the role of Sonic Hedgehog signaling in 

establishing midbrain dopaminergic neuron subclasses 

Anna Kabanova, Sandra Blaess 

Neurodevelopmental Genetics, Institute of Reconstructive Neurobiology, Life and Brain Center, University 

of Bonn, Sigmund-Freud-Str. 25, D-53127 Bonn. 

The midbrain dopaminergic (mDA) neurons of the substantia nigra (SN) and 

the ventral tegmental area (VTA) play critical roles in the control of voluntary 

movement, and reward behavior, respectively. These subpopulations of mDA 

differ in gene expression, axonal projections and their aberrant function underlies 

a wide spectrum of disorders, such as Parkinson’s disease and schizophrenia. 

All mDA neurons appear to be derived from a precursor domain at the ventral 

midline of the developing midbrain. Sonic Hedgehog (Shh) signaling is necessary 

for the induction of the mDA precursor domain, but it remains unclear whether 

Shh plays additional roles in mDA specification and differentiation at subsequent 

stages of development. 

To investigate later roles of Shh signaling in mDA neurogenesis, we are analyzing 

mutants in which Gli2, the main activator downstream of Shh signaling, 

was removed in the vMb between embryonic day (E) 8.5-E9.0. In the mutant 

mice, we observed a severe reduction in the number of mDA neurons in the 

adult midbrain. Evaluation of the expression of mDA subset markers and mDA 

projections to the forebrain showed that the majority of the remaining mDA 

neurons in the mutants adopt the fate of neurons in the SN, whereas only a small 

number of neurons are specified to the mDA neurons of the VTA. Analysis of the 

developing midbrain demonstrates that mDA neurons are already reduced and 

disorganized at E11.5. Together with fate mapping studies that show that mDA 

precursors that give rise to the ventral-medial VTA respond to Shh signaling after 

E9.0, these results indicate that in addition to an early role in mDA precursor 

induction, Shh signaling could influence the specification and fate decision of 

mDA neurons. Using conditional mosaic inactivation, we are currently addressing 

whether Shh signaling plays a direct role in this process.

64 


Local gene targeting and cell positioning using 

magnetic nanoparticles for the generation of 

biological cardiac pacemakers 

Carsten Kilgus, Tobias Bruegmann, Bernd K. Fleischmann and Philipp Sasse 

Institute of Physiology I, Life&Brain Center, University Bonn, Sigmund-Freud-Str. 25, 53105 Bonn, Germany. 

Implantation of electronic implantable pacemakers is the primary therapy for 

patients with sinus node dysfunction or high degree of atrio-ventricular block. 

Although complications are rare, this treatment has some limitations which is 

why the development of biological pacemakers is studied. A biological pacemaker 

should be confined to an area within the heart and we have therefore developed 

new technologies for localized cell therapy and gene transfer to cardiomyocytes 

using magnetic nanoparticles (MNP) and magnetic fields. 

First initial proof of principle studies, a monolayer cardiomyocytes from the HL-1 

line transfected with a lentivirus that expressed the green fluorescent protein 

(GFP). Localization was achieved by generation of complexes with MNP and 

lentiviruses (200 fg iron per virus particle) and transduction of cells in a confined 

magnetic field that was generated by neodymium-iron-boron permanent 

magnets with specially designed soft iron tips. This approach allowed reliably 

and stable transduction of cells within an area of < 1mm diameter indicated by 

localized GFP expression. 

To provide a functional pacemaker we plated HL-1 cells onto multi-electrode 

arrays (MEA, 60 electrodes, 200 µm spacing) to read out the local field potentials 

and calculated the site from where pacemaking was initiated. We performed 

localized transduction with MNP-lentivirus complexes that express the light-gated 

ion channel channelrhodopsin2 (ChR2). This lead to localized ChR2 expression 

and light flashes paced the whole monolayer. Importantly latency analysis of the 

MEA recordings proved that pacing was always initiated from the transduced sites 

despite global illumination. To be able to generate an autonomeous pacemaker, 

we have successfully generated lentiviruses to express the classical pacemaker 

channels HCN2 and HCN4 as well as a dominant-negative potassium channel 

(Kir2.1-AAA). Preliminary long-term MEA recordings and localized transduction 

with the Kir2.1-AAA virus indicated that pacemaking is initiated from the 

transduced site 2-3 days after localized gene transfer. 

Besides gene transfer we have also tested localized cell positioning for pacemaker 

generation. Therefore we have differentiated and purified ChR2-expressing

65 


cardiomyocytes from embryonic stem cells, which could be paced by light. 

These were loaded with MNP, guided onto HL-1 monolayers on MEA by localized 

magnetic fields. By this approach only 5000 cells were needed to generate a 

confined cell cluster (~ 1mm diameter). Importantly, global light stimulation 

could be used to pace the monolayer of HL-1 cells and pacing started from the 

guided ChR2-cell cluster. 

In summary, magnetic nanoparticles and localized magnetic field are very 

effective tools for localized gene transfer and cell positioning to generate cardiac 

pacemakers in vitro. The application of this technology for light-induced or 

autonomic cardiac pacing in vivo is currently under investigation.

66 


Abnormal functional connectivity between areas 

involved in emotion and executive control in 

psychogenic non-epileptic seizures (PNES) 

S.J.M. van der Kruijs 1,2 , N.M.G. Bodde 1 , M.J. Vaessen 1,2 , R.H. Lazeron 1 , P.A. 

Hofman 1,2 , W.H. Backes 2 , A.P. Aldenkamp 1,2 , J.F.A. Jansen 2 

1 Epilepsy Center Kempenhaeghe, Heeze, Netherlands; 2 Department of Radiology, Maastricht University 

Medical Center, Maastricht, Netherlands. 

Introduction 

Psychogenic non-epileptic seizures (PNES) are paroxysmal episodes without 

epileptiform brain activity as recorded by EEG. Instead, there is positive 

evidence for psychogenic factors that may underlie the seizures. One important 

predisposition factor for PNES is dissociation, i.e. disrupted integration of 

conscious functioning. We explored whether fMRI could identify biomarkers of 

brain alterations associated with dissociation in PNES. 

Methods 

Patients with PNES (n=11) and healthy volunteers (n=12) completed dissociation 

questionnaires (DIS-Q, DES and SDQ-20), and the Raven’s Progressive Matrices 

Test. The fMRI protocol consisted of 4 scans: Resting state fMRI (rsfMRI) 1, 

picture encoding, Stroop color naming, and rsfMRI 2. All MR images were preprocessed 

in SPM8, and between-group differences in cerebral activation were 

assessed. Subsequently, ROIs with strong activation were defined, based on 

the activation patterns during the tasks, averaged over all subjects. Based on 

the ROIs, functional connectivity maps were obtained from resting-state fMRI. 

Multiple regression was performed to assess between-group differences (p

67 


Conclusion 

The abnormal strong functional connectivity in PNES patients provides a 

neurophysiological correlate for an underlying dissociation mechanism where 

emotion can result in altered motor control, resulting in seizure-like episodes.

68 


Characterisation of two N-acetylaspartylglutamate 

synthetases 

J. Lodder-Gadaczek, M. Eckhardt, V. Gieselmann 

Institut für Biochemie und Molekularbiologie, Universitätsklinikum Bonn. 

N-acetylaspartylglutamate (NAAG) is an abundant neuropeptide in the 

mammalian brain, present in micromolar to millimolar concentrations. NAAG 

is synthesized in specific neurons by a synthetase catalyzing a condensation of 

N-acetylaspartate (NAA) and glutamate. After release from synaptic terminals 

NAAG can be degraded by glutamate carboxypeptidase II (GCPII) or glutamate 

carboxypeptidase III (GCPIII). The released products can then be transported to 

oligodendrocytes and astrocytes. 

We and others (Collard, 2010) identified the NAAG synthetase-I (NAAGS-I) 

encoded by a ribosomal modification protein rim-like family member B (Rimklb) 

gene and the NAAG synthetase-II (NAAGS-II) a structurally related protein 

encoded by a ribosomal modification protein rim-like family member A (Rimkla) 

gene. Besides NAAGS-II`s production of NAAG we determined the synthesis of an 

acetylated tripeptide N-acetylaspartylglutamylglutamate (NAAG 2 ), which is not 

synthesized by NAAGS-I. 

Tandem mass spectrometric measurements showed NAAG 2 presence in 

nanomolar concentrations in the murine nervous system, with the highest 

concentrations in sciatic nerv, spinal cord and stem brain, which correlate with 

the expression levels of NAAGS-II. The NAAGS-II knock-out mice showed no 

NAAG 2 . 

Furthermore we found proof that NAAG and NAAG 2 react in the same metabolic 

system. 

This is the first description of the presence of NAAG 2 in the vertebrate nervous 

system. For now the physiological roles of NAAG 2 are not understood and remain 

to be determined.

69 


Local overexpression of Interleukin -11 in the central 

nervous system prevents demyelination 

A. Maheshwari 1 , H. Slaets 1 , C. Van den Haute 2 , V. Baekelandt 2 , P. Stinissen 1 , 

J. Hendriks 1 , N. Hellings 1 

1 Hasselt University, Biomedical Research Institute,and transnationale Universiteit Limburg, School of Life 

Sciences, Diepenbeek, Belgium; 2 Katholieke Universiteit Leuven, Laboratory for Neurobiology and Gene 

Therapy, Division of Molecular Medicine, Leuven, Belgium. 

Background & Aim 

Demyelination is one of the clinical hallmarks of multiple sclerosis (MS). Till 

today no therapy is available which potentiates the endogenous remyelination. 

Interleukin-11 (IL-11) is a member of the gp130 family cytokines, that also 

includes leukemia inhibitory factor and oncostatin M. These neuropoietic 

cytokines are upregulated in MS lesions and several have been shown to control 

neuroinflammation. Recently, IL-11 was identified to directly provide trophic 

support to oligodendrocytes by potentiating their survival and maturation in 

vitro. Moreover, IL-11 was shown to improve clinical severity in the EAE model. 

In this study we aim to elucidate the direct in vivo effect of IL-11 demyelination 

independent of the immune response. 

Methods 

To achieve immune independent demyelination, a cuprizone induced 

demyelination mouse model was used. 0.2% of cuprizone was mixed with 

powdered chow and fed to 8 week old mice for 5 weeks to achieve acute 

demyelination. To allow spontaneous remyelination, mice were returned to 

normal diet after 5 weeks. CNS directed overexpression of IL-11 was achieved by 

stereotactical administration of IL-11 encoding lentiviral vectors in the striatum of 

mouse brain. For the prophylactic study, vectors were administered 2 weeks prior 

to the start of the cuprizone diet and acute demyelination was studied. For the 

therapeutic study, the vectors were administered 4 weeks after the start of the 

cuprizone diet. Mice were allowed to remyelinate for 2 weeks and subsequently 

sacrificed for immunohistochemical analyses (Luxol Fast Blue, Iba-1, NG-2). 

To study the in vitro effect of IL-11 on myelin phagocytosis, RAW264.7 cells were 

cultured and treated with 1, 3, 10, 30 and 100 ng/ml of murine rIL-11 for a period 

of 6 and 12 hrs. The uptake of DI labeled myelin was analysed by flowcytometry. 

Results 

Overexpression of IL-11 in mouse brain striatum significantly limited the area 

of acute demyelination (decrease of 27% as compared to controls) in the

70 


area of corpus callosum around the midline. Moreover, the number of NG2 + 

oligodendrocyte progenitor cells (OPC) and Iba-1 + microglia were found to be 

significantly reduced compared to controls. In vitro, IL-11 dose dependently 

increased myelin phagocytosis (1, 3, 10, 30 ng/ml) by RAW 264.7 cells. Currently, 

the effect of therapeutic application of IL-11 is being evaluated. 

Conclusion 

Prophylactic delivery of IL-11 reduces acute demyelination in the cuprizone model. 

IL-11 potentiates myelin phagocytosis in vitro, which could point to enhanced 

clearing of myelin debris in vivo thereby creating an environment conducive for 

repair processes in the demyelinated area. Moreover, the reduced number of 

OPC may indicate that differentiation into mature myelinating oligodendrocytes 

has occurred, which then play their putative role in remyelination. Future studies 

will have to confirm these findings.

71 


Identification of microRNAs regulating neuronal 

differentiation of human neural stem cells 

Laura Mürtz 1 , Lodovica Borghese 1 , Beate Roese-Koerner 1 , Sandra Weinhold 2 , 

Philipp Koch 1 , Peter Wernet 2 , Markus Uhrberg 2 and Oliver Brüstle 1 

1 Institute of Reconstructive Neurobiology, Life and Brain Center, University of Bonn; 2 Institute for Transplantation 

Diagnostics and Cell Therapeutics, Heinrich-Heine University Düsseldorf. 

MicroRNAs are a class of small non-coding RNAs that act as post-transcriptional 

regulators of gene expression. Studies in model organisms have demonstrated 

critical roles for a number of microRNAs in neuronal development, but their 

specific functions have not been fully examined. Likewise, data on microRNA 

function in the context of human neural development and neural stem cell 

biology is particularly scarce. Our human embryonic stem cell (hESC) based 

neural differentiation paradigm provides a useful model for the analysis of 

microRNA profiles in human pluripotent and multipotent stem cells and for the 

identification and functional characterization of novel microRNAs associated 

with neuronal differentiation. We assessed microRNA of human ES cells (hES, 

I3 line), of long-term self-renewing neuroepithelial-like stem cells (lt-NES® cells) 

(Koch et al. 2009) and of 15- and 30-days old differentiated neuronal cultures 

(ND15, ND30) using a quantitative real-time PCR multiplex assay. We validated 

the identified expression patterns for several microRNAs by Northern blot 

analysis in two independent hES cell lines (I3 and H9.2) indicating the reliability 

of our approach. Furthermore we were able to confirm previous assignments of 

ES cell-specific and brain-specific microRNAs. In addition, we identified several 

novel microRNAs, besides the known neuronal microRNAs, i.e. miR-124 and 

miR-125, as up-regulated during human neural stem cell differentiation. Stable 

overexpression of the newly identified microRNAs, just like the overexpression of 

miR-124 and miR-125, resulted in an increased rate of neuronal differentiation. Our 

data demonstrate that these microRNAs are not only markers of differentiation 

but may also function as regulators of human neuronal differentiation.

72 


Role of Preproenkephalin (PENK) gene in stress 

reactivity 

Irene Melo de Carvalho 

Institute of Molecular Psychiatry, University of Bonn, Germany. 

Stress reactivity is a response of the organism that aims to replace an homeostatic 

state after internal or external stimuli. Diverse physiological components are 

implicated in the stress reactivity, at which the activation of the HPA axis seems to 

be crucial. The HPA axis is composed of the hypothalamic paraventricular nucleus 

and the pituitary and adrenal glands. These structures release the neuronal 

factor CRF and the hormones ACTH and glucocorticoids. These hormones are key 

factors in the activation of other components involved in stress reactivity. 

Severe or chronic stressful stimuli can lead to an abnormal activation of the HPA 

axis, leading to alterations in diverse brain and peripheral pathways associated 

with stress disorders like phobias, chronic anxiety states or depression. 

Recently, the opiod peptide enkephalin, which is encoded by the PENK gene, has 

been related to stress reactivity, mainly in studies with PENK knockout animals. 

Under basal and acute stress situations, these animals showed augmented stress 

reactivity. 

Our aim is to analyze the role of the Penk gene and enkephalin peptide in stress 

reactivity under chronic stress situations, in behavioral experiments and related 

cellular and molecular pathways. 

Up to know, we found that Penk knockout animals exposed to chronic mild stress, 

an animal model of anxiety and depressive like states, show less susceptibility to 

chronic stress than WT animals. Penk KO animals, in contrast to WT, exhibited no 

anxiety phenotype in O-maze test and no despair behavior in Porsolt test. 

These results suggest that exacerbated reaction under acute stress situations 

in PENK KO animals can have a protective role in chronic stress. Experiments 

to understand the cellular and molecular mechanism beyond this possible 

protective effect are ongoing.

73 


The role of Goodpasture antigen-binding protein 

(GPBP) in the cellular response against Aβ-induced 

toxicity 

Chiara Mencarelli 1 , Gerard H. Bode 1 , Marjorie Gangolf 1 , Sonia Tosheva 2 , Jochen 

Walter 2 , Mario Losen 1 , Peter Molenaar 1 , Harry W.M. Steinbusch 1 , Marc H. De 

Baets 1 , and Pilar Martinez-Martinez 1 

1 Department of Neuroscience, School of Mental Health and Neuroscience, Maastricht University, Maastricht, 

the Netherlands; 2 Department of Neurology, University of Bonn, Bonn, Germany. 

Alzheimer’s disease (AD) is a complex multifactorial syndrome which is 

characterized by intra- and extracellular deposits of amyloid beta (Aβ), 

neurofibrillary tangles, dystrophic dendrites and increased levels of ceramide 

in the cerebrospinal fluid. Goodpasture antigen-binding protein (GPBP) is an 

ubiquitous protein which can bind to proteins that are prone to misfold and 

aggregate. GPBP is widely distributed in the CNS, where it is involved in brain 

homeostasis and development. We hypothesize that GPBP plays a role in the 

pathogenesis of neurodegenerative diseases that are characterized by the 

formation of protein aggregates. Secreted GPBP forms extracellular complexes 

with Aβ. We found that GPBP accumulates in amyloid plaques from AD patients 

and colocalizes with Amyloid P component and Aβ fibrils. GPBP gene and protein 

levels are increased in HEK cells over-expressing APP Sw . Finally we found that 

GPBP reduces Aβ aggregation and protects against Aβ-induced toxicity in cultured 

neurons. Our results indicate that GPBP takes part in the cellular response 

against Aβ-induced toxicity and thus is may be involved in the pathophysiology 

of neurodegenerative processes.

Investigating the role of microRNAs in 

Spinocerebellar Ataxia type 3 

Rohit Nalavade 

74 


DZNE, German Center for Neurodegenerative Disases, Ludwig-Erhard-Allee 2, 53175 Bonn, Germany 

Spinocerebellar Ataxia type 3 (SCA3) is an autosomal, dominantly inherited 

disorder belonging to the group of polyglutamine repeat (polyQ) diseases. It is 

caused by CAG repeat expansions in the SCA3 gene coding for polyglutamine 

repeat expansions in the Ataxin-3 (At3) protein. The disease is characterized by 

intranuclear aggregates and progressive neuronal cell death especially in the 

cerebellum and the brain stem. MicroRNAs (miRNAs) have been shown to play 

important roles in neurodegenerative diseases. This study aims to investigate 

if miRNAs play a role in regulation of At3 protein expression. MiRNA profiling 

and target prediction software have suggested the involvement of miRNAs in 

regulation of At3 expression. Further experiments using constructs including 3’ 

Un-Translated Region (UTR) of At3 mRNA coupled with luciferase and lentiviral 

constructs for over-expression of these miRNAs have backed the initial results 

suggesting that some of these miRNAs may indeed play a role. A promising 

recent development has been the use of neurons derived from IPSCs (Induced 

Pleuripotent Stem Cells) from patients of SCA3 as a system to study various 

aspects of the disease. Assessment of the miRNA profile of these neurons and 

their further use in experiments involving modulations of specific miRNA levels is 

expected to provide vital insights regarding the role of miRNAs in SCA3.

75 


Development of an in vitro system for the 

assessment of axon growth promoting properties of 

bioengineered scaffolds 

Gerardo-Nava J 1,2 , Grehl T 3 , Weis J 2 , Steinbusch HWM 1 , Brook GA 2 . 

1 Maastricht University, School of Mental Health and Neurosciences; 2 RWTH Aachen University Hospital, 

Institute of Neuropathology; 3 Ruhr-University-Bochum, Department of Neurology. 

An in vitro model capable of studying the axon growth promoting properties of 

3D biomaterials will be useful in the development of scaffolds for the repair of 

traumatically injured peripheral nervous system (PNS) tissues. The well preserved 

cytoarchitectural organization of spinal cord organotypic slice cultures (SCOSC) 

as well as hemisected dorsal root ganglia (DRG) are being used to determine 

the orientated motor- and sensory axon growth promoting properties of a 

3D collagen scaffold. Sensory neurons from hemisected DRGs extended long 

and highly orientated SMI32-positive axons throughout the Matricel collagen 

microchannels as well as along the surface of the scaffold. Axons that reached 

the dorsal horn of the SCOSC crossed the scaffold-SCOSC interface and matrix 

crossing it and exploring the target tissue of the DH of the organotypic slice 

cultures. Similarly, ventral horn motor neuron axons (also SMI-32 positive) from 

the of slice cultures extend within and along the surface of the scaffold. In both 

cases, a close interaction between regenerating axonal profiles and NGFr/p75postive 

Schwann cells could be seen. Cell migration into the collagen scaffold 

was strong in both experimental set-ups and included migration by cells that 

were immunoreactive for NGFr/p75, S100, IBA1 and ED1. A more limited degree 

of GFAP-positive astrocyte migration was observed close to the SCOSC-scaffold 

interface. Earlier in vitro investigations have focused largely on cell-substrate 

interactions using cell suspensions and 2D biomaterials. As the field of biomaterial 

research advances in the development of 3D scaffolds, the utilization of in vitro 

models capable of analyzing the tissue-scaffold interactions may generate much 

useful information. In particular, it is hoped that such approaches will eventually 

reduce the number of animals required in the development of bioengineering 

strategies intended for PNS repair.

76 


Presenilins regulate induction of autophagy 

Nguyen Thanh Tien 

Department of Neurology, University of Bonn, Germany 

Introduction 

Macroautophagy or autophagy, is a lysosome-dependent degradative pathway 

for organelles and nutrient recycling. The process starts with formation of 

autophagosomes, which then fusing with lysosomes to become autolysosomes. 

Autophagy has also been implicated in the Alzheimer’s disease (AD) associated 

neurodegeneration. Sequential cleavage of β-amyloid precursor protein by β- 

and γ- secretases generates β-amyloid (Aβ), which deposits as amyloid plaques 

in the AD brains. Mutations in presenilins (PS1/PS2), the catalytic subunits of 

γ-secretase complex, are major cause of early-onset familial AD. Recent studies 

show enrichment of PS1 in autophagic vacuoles, and also indicate potential role 

of PS1/PS2 and PS1 homolog in regulation of autophagic flux. 

Aims 

We address the role of presenilins in autophagy in steady-state and adaptive 

response to stimuli, and possible involvement of the class III phosphoinositide 

3-kinase (PI3K) complex in this process. 

Methods 

PS knock-out and PS1 expressing mouse embryonice fibroblasts and human 

embryonic kidney cells were used as experimental model. Autophagic flux 

was induced by starvation or impaired by lysosomal inhibitors. The changes of 

autophagy were assessed by biochemical analyses. 

Results 

LC3 is widely used as a major marker to assess autophagic flux. LC3-I, a 

cytosolic form, is lipidated to LC3-II, which is anchored into the membrane of 

autophagosomes. At steady state, western blot analysis shows accumulation of 

both LC3-I and LC3-II in PS1 wild-type (PS1wt) expressing cells compared to PS 

deficient cells. Immunocytochemistry also prove abundance of LC3 puncta per 

cells in PS1wt expressing cells in comparison to PS deficient cells. Upon short 

starvation, ratio of LC3-II/LC3-I is increased in PS1wt expressing cells as compared 

to PS deficient cells.

77 


Cathepsin D, a lysosomal aspartyl protease, is used to assess lysosomal 

degradation. However, the ratio of active Cathepsin D/pre-pro Cathepsin D was 

not change in cells expressing PS1 variants. 

Beclin 1 is an essential component in the PI3K class III complex to regulate 

autophagic induction. It associates with bcl-2 – an antiapoptotic protein to inhibit 

autophagosome build-up under normal nutrient condition. Phosphorylation of 

either beclin 1 or bcl-2 results in dissociation of this complex and the activation 

of autophagy. Western blotting demonstrates that beclin 1 and bcl-2 levels do 

not change upon PS1 variants. Interestingly, phosphorylation of bcl-2 (Ser 70) 

significantly increases in PS1wt expressing cells as compared to PS deficient 

cells. 

Conclusion 

Our data indicate that PS regulates the induction of autophagy in steady-state 

and during starvation. PS1wt expressing cells show increases in the size and/ 

or number of autophagic vacuoles in comparison to PS deficient cells. This 

accumulation is associated with phosphorylation of bcl-2 at Ser 70. 

The involvement of PS in autophagy could also contribute to degradation of AD 

- associated proteins

78 


Effect of prenatal stress and developmental fluoxetine 

exposure on hippocampal glucorticoid receptors and 

coactivator GRIP1 in adolescent offspring 

N.A. Niessen 1,3 , J. L. Pawluski 1,2,3 , I. Rayen 2,3 , D.L. van den Hove 2,3 , H.W.M. 

Steinbusch 2,3 , J. Balthazart 1,3 , and T.D. Charlier 1,3 

1 GIGA-Neurosciences, University of Liège, Liège, Belgium; 2 Department of Neuroscience, Maastricht University, 

Maastricht, The Netherlands; 3 EURON, European Graduate School for Neuroscience. 

Up to 20% of women suffer from depression during pregnancy and the 

postpartum period. Selective serotonin reuptake inhibitors (SSRIs) are commonly 

used to treat maternal depression and up to 10% of mothers are prescribed this 

type of medication. While maternal stress, depression and anxiety have longterm 

effects on the physical and behavioral development of offspring, numerous 

preclinical studies also point to a significant action of developmental exposure to 

SSRIs. Surprisingly, data are limited concerning the combined effect of maternal 

depression and maternal SSRI exposure on offspring development. Our goal was 

to determine how maternal fluoxetine exposure affects the neurodevelopment of 

male and female adolescent offspring in an animal model of maternal adversity. 

Because previous studies suggest that exposure to SSRI affects the HPA axis 

during adulthood, we studied the expression of glucocorticoid receptors (GR), 

mineralocorticoid receptors (MR), and the associated steroid receptor coactivator 

GRIP1 in the hippocampus, a brain region sensitive to stress. Gestationally 

stressed and non-stressed Sprague-Dawley rat dams were chronically treated 

throughout lactation with either fluoxetine (5mg/kg/day) or vehicle beginning 

on postnatal day 1. We therefore obtained four groups of male and female 

offspring: 1) prenatal stress + fluoxetine exposure, 2) prenatal stress + vehicle, 

3) fluoxetine exposure alone, and 4) vehicle alone. Offspring were weaned at 

P21 and killed during adolescence, at P42. Our findings show that developmental 

fluoxetine exposure leads to an overall decrease in GR immunoreactivity, but not 

MR expression, in the hippocampus of adolescent offspring, independently of 

prenatal stress. Interestingly, this effect was more pronounced in males. We are 

currently analyzing the expression pattern of GRIP1 throughout the hippocampus. 

Altogether, these results highlight a direct effect of developmental exposure to 

SSRI medications on the capacity of the hippocampus to respond to stress during 

adolescence.

79 


Cortical abnormalities and language impairment in 

Rolandic epilepsy 

*G.M. Overvliet 1,2,3 , *R.M.H.Besseling 1,2,4 , A.P. Aldenkamp 1,2,3 J.F.A Jansen 2,4 , J.S.H. 

Vles 1,5 , S. Ebus1 P. Hofman 4,2 , W.H. Backes 4,2 

1 Epilepsy center Kempenhaeghe, Heeze, the Netherlands; 2 Research school of Mental Health & Neuroscience, 

Maastricht University, Maastricht, the Netherlands; 3 Department of Neurology. Maastricht University 

Medical Center, Maastricht, the Netherlands; 4 Department of Radiology, Maastricht University 

Medical Center, Maastricht, the Netherlands; 5 Department of Child Neurology, Maastricht University 

Medical Center, Maastricht, the Netherlands. 

* Both authors contributed equally in this study 

Background 

In clinical practice, children with Rolandic epilepsy may display language 

impairment. Epileptiform discharges have been associated with disturbed 

functionality of the language network. However, whether abnormalities in 

cortical morphology exist and may explain the language impairment has not 

been investigated. The aim of this study is to compare the cortical thickness and 

language performance of children with Rolandic epilepsy with healthy controls. 

Methods Cross-sectional study of children with Rolandic epilepsy. Structural 

T1-weighted MRI (1 mm iso, 3 Tesla) and the Clinical Evaluation of Language 

Fundamentals (CELF-4, Dutch edition) language test were performed in 21 

children with typical Rolandic epilepsy confirmed by EEG (age, mean±SD: 

135±24 months) and 21 age-matched controls (125±20 months). The Freesurfer 

image analysis software package was used to measure cortical thickness and to 

perform linear regression with CELF language metrics and age. Cortical regions 

of abnormal thickness were identified using uncorrected trends of p

80 


Conclusions 

For the first time, abnormal cortical morphology was identified in children with 

Rolandic epilepsy relative to age-matched healthy controls. The observed cortical 

thinning was localized in language mediating brain regions in the left hemisphere, 

correlated with age. This probably suggest a growing into deficit phenomena in 

Rolandic epilepsy. These observations suggest that Rolandic epilepsy cannot be 

merely considered as a benign condition in children.

81 


Optogenetic modulation of the septohippocampal 

pathway 

Milan Pabst 


Germany. 

The septohippocampal projection is thought to play a critical role in the 

generation and the maintenance of various forms of rhythmic activity in the 

hippocampus, i.e. theta rhythm. Septal neurons projecting to the hippocampus 

can be subdivided into GABAergic and cholinergic neuronal subtypes, with 

distinctive axonal morphology and terminations. However, the precise functional 

effects of stimulating GABAergic or cholinergic septohippocampal afferents on 

hippocampal neuronal networks have remained unclear, primarily because of the 

difficulty of stimulating septohippocampal fibers selectively. We have expressed 

channelrhodopsin-2 (ChR2) in either cholinergic or GABAergic septal neurons 

using recombinant adeno-associated virus (rAAV) mediated gene transfer of a 

construct leading to Cre-dependent expression of ChR2 in mouse lines expressing 

Cre recombinase only in cholinergic (ChAT-Cre) or GABAergic (PV-Cre) septal 

neurons. We have examined the effects of optogenetic stimulation of cholinergic 

projection axons on hippocampal microcircuits in the dentate gyrus.

82 


Does the proteoglycan NG2 influence neuron-NG2 cell 

synaptic signaling? 

S. Passlick 1 , K. Karram 2,3 , J. Trotter 3 , G. Seifert 1 , C. Steinhäuser 1 , R. Jabs 1 

1 Institute of Cellular Neurosciences, University of Bonn, Germany; 2 Institute for Molecular Medicine, University 

of Mainz, Germany; 3 Molecular Cell Biology, University of Mainz, Germany. 

Glial cells expressing the proteoglycan NG2 are widely distributed throughout 

the developing and adult gray and white matter of the CNS. Several properties 

distinguish them from astrocytes, mature oligodendrocytes and microglia. NG2 

cells express different types of voltage-gated K + , Na + , and Ca 2+ -channels. They 

also express a variety of ligand-gated receptors including group I metabotropic 

glutamate receptors and ionotropic AMPA- and GABA A -receptors. Furthermore, 

NG2 cells are the only non-neuronal cells in the CNS that form synapses with 

neurons. In this respect, it is interesting that the NG2 protein (i) binds to the 

postsynaptic Glutamate Receptor Interaction Protein (GRIP) and (ii) contains 

two Laminin G/Neurexin/Sex Hormone Binding Globulin (LNS) domains in the 

extracellular region. GRIP is considered important for clustering of the GluR2 

subunit of AMPA receptors. LNS domains are characteristic for postsynaptic 

neurexins that, by binding to presynaptic neuroligins, are important for synapse 

formation in neurons. 

In this study we asked whether the NG2 protein is crucial for the formation of 

functional NG2 cell synapses, by influencing clustering of postsynaptic receptors 

or neuroligin interactions. To address this issue, we investigated synaptic 

transmission between glutamatergic neurons and NG2 cells in NG2-EYFP-knockin 

(+/- and -/-) and wildtype mice (p 8-14). We recorded whole-cell membrane 

currents from hippocampal NG2 cells during electrical stimulation of Schaffer 

collaterals and analysed the evoked excitatory postsynaptic currents (eEPSCs). 

Comparison of the kinetics and paired-pulse ratios of NG2 cell eEPSCs revealed 

no significant differences among the tested genotypes. 

We conclude that the lack of the NG2 protein does not cause a general failure 

of synaptic signaling between glutamatergic neurons and NG2 cells in the 

hippocampus. It remains to be tested whether miniature EPSCs, which are not 

synchronised by presynaptic action potentials, are affected in NG2-deficient 

mice. 

Supported by DFG (SPP 1172) and EC (FP7-202167 Neuroglia).

83 


Multiple Mitochondrial DNA deletions in human 

disease 

Viktoriya Peeva, Gábor Zsurka, Susanne Schöler, Alexei P. Kudin, Stefan Vielhaber, 

Grazyna Debska-Vielhaber, Werner Zuschratter, Cornelia Kornblum, and Wolfram 

S. Kunz 

Division of Neurochemistry, Department of Epileptology and Life&Brain Center, University Bonn, 

Sigmund-Freud-Str. 25, 53105, Bonn, Germany. 

Charcot-Marie-Tooth neuropathy type 2A (CMT2A) is an autosomal dominant 

axonal form of peripheral neuropathy caused by mutations in the mitofusin 2 

gene (MFN2), which encodes a mitochondrial outer membrane protein that 

promotes mitochondrial fusion. In this study we investigated two novel MFN2 

mutations and their effect on mitochondrial function in skeletal muscle and 

cultured fibroblasts of two patients affected by the disease. 

To elucidate the potential mitochondrial impairment in skeletal muscle biopsies 

and fibroblasts of these patients, high-resolution respirometry and sensitive PCRbased 

deletion detection methods were applied, and the mtDNA copy number 

was determined. 

Altered distribution of mitochondria in type 2A skeletal muscle fibers, 

characterized with accumulation of subsarcolemmal mitochondria and 

rarefication of intermyofibrillar mitochondria were observed in skeletal muscle 

biopsy from the patients. 

The sensitivity of saponin-permeabilized muscle fibers and digitoninpermeabilized 

fibroblasts to the cytochrome c oxidase (COX) inhibitor azide 

increased, although their maximal activities of respiration were only slightly 

affected by the MFN2 mutations. 

In comparison to controls, the MFN2 muscle showed decrease in the mtDNA copy 

number (the extent of mtDNA depletion was lower when compared to patients 

carrying POLG mutation), and elevated levels of mtDNA deletions. Overall, these 

mtDNA changes are implicated to cause the increased flux control of COX in 

CMT2A fibroblasts and skeletal muscle. 

Our findings support the viewpoint that impairment of the fusion proteins affect 

mitochondrial function. This appears to be related to the role of mitochondrial 

dynamics for segregation of mtDNA mutations.

84 


Effects of anticonvulsant drugs on hippocampal 

inhibitory microcircuits in the epileptic hippocampus 

Leonie Pothmann, Heinz Beck 


Germany. 

The anticonvulsant carbamazepine (CBZ) acts via inhibition of voltage-gated 

Na + channels, showing an increase of inhibition when neurons fire at high 

frequencies. This use-dependent block is thought to be important in mediating 

the inhibition of pathological high-frequency seizure activity. However, some 

inhibitory interneuron types are capable of firing at very high rates, suggesting 

that CBZ should cause impaired GABAergic inhibition, and consequently increase 

excitability. 

Therefore, we examined the effects of CBZ on different cell types in the rat 

hippocampus. We found that pyramidal cell firing was inhibited by CBZ, as 

expected. However, interneurons that predominantly target pyramidal cell 

somata (group I), and interneurons targeting the apical dendritic tuft of 

pyramidal neurons (group II) did not show significant effects of CBZ. A third 

group of interneurons innervating the proximal dendrites of pyramidal cells in 

stratum oriens and stratum radiatum was affected by CBZ, showing a decrease 

of 44 % in maximal firing rate. To determine the impact of this reduction on 

inhibitory micronetworks we recruited feed-back and feed-forward inhibition 

of CA1 pyramidal cells by electrical stimulation of CA1 and CA3 pyramidal cell 

axons, respectively. At stimulation frequencies of 50 Hz, neither feed-back nor 

feed-forward inhibition was disturbed. 

In the pilocarpine model of chronic epilepsy we found marked changes in the 

responsiveness of different cell types: pyramidal cells showed a significant 

decrease of 14 % in the response to CBZ, whereas group I interneurons that 

where unresponsive in control animals now showed a block of 40 % in maximal 

firing rate. In contrast the pronounced effect on group III interneurons was lost in 

epileptic animal whereas the resistance of group II interneurons was unchanged. 

These data suggest differences in the properties of Na+ channels of different cell 

types, and that epileptogenesis leads to a marked change in the network effects 

of anticonvulsant drugs.

85 


Maternal fluoxetine exposure, regardless of prenatal 

stress, affects physiological systems involved in sexual 

development of offspring 

Ine Rayen 1 , Thierry D. Charlier 2 , Stephanie Mollegaard Kristensen 1 , Jacques 

Balthazart 2 , Harry Steinbusch 1 , Jodi L. Pawluski 1,2 

1 School for Mental Health and Neuroscience, Maastricht University, Universiteitssingel 50, 6229 ER 

Maastricht, The Netherlands; 2 University of Liege, GIGA-Neurosciences, 1 avenue de l'Hôpital (Bat. B36), 

B-4000 Liège, Belgium. 

Depression during pregnancy and postpartum is a growing health problem and 

affects up to 20% of women. While selective serotonin reuptake inhibitor (SSRIs) 

medications are commonly used for treatment of maternal depression, the 

combined effect of maternal depression and perinatal SSRI exposure on offspring 

development is poorly investigated. Here, our aim was to determine the role of 

exposure to fluoxetine during development on sexual behavior in offspring using 

a rodent model of maternal adversity. To do this, gestationally stressed and nonstressed 

Sprague-Dawley rat dams were chronically treated throughout lactation 

with either fluoxetine (5mg/kg/day) or vehicle beginning on postnatal day 1 (P1). 

Four groups of male and female offspring were obtained: 1) prenatal stress + 

fluoxetine exposure, 2) prenatal stress + vehicle, 3) fluoxetine exposure alone, 

and 4) vehicle alone. In Experiment 1, we assessed ano-genital (AG) distance 

in juvenile male and female offspring. In Experiment 2, adult male and female 

offspring were tested to assess the effect of developmental exposure to fluoxetine 

and prenatal stress on sexual behavior. Maternal fluoxetine exposure significantly 

decreased AG distance in juvenile male offspring, regardless of prenatal stress. 

AG distance in juvenile female offspring was not affected. In adult male offspring, 

preliminary results demonstrate that maternal fluoxetine treatment, regardless of 

exposure to prenatal stress, significantly decreased the number of intromissions, 

significantly increased the latency to first intromission and tended to increase 

the latency to first ejaculation. Further work will investigate the effect of prenatal 

stress and developmental fluoxetine exposure on sexual behavior in adult female 

offspring. We will also investigate the neurobiological plasticity underlying these 

behavioral effects. These preliminary results provide important evidence of the 

long-term impact of maternal fluoxetine use on the development of fundamental 

physiological systems.

86 


The impact of bifunctional microRNA-9/9* on the 

differentiation of human ES cell – derived neural stem 

cells 

Beate Roese-Koerner, Lodovica Borghese, Laura Mürtz, Oliver Brüstle 

Institute for Reconstructive Neurobiology, University of Bonn. 

MicroRNAs are non-coding RNA molecules about 22 nucleotide-long that 

regulate gene expression at a post-transcriptional level. MicroRNAs are known 

to be involved in many cellular processes including stem cell self-renewal and 

differentiation. We carried out a comparative microRNA expression profiling 

across the differentiation of human embryonic stem cells (hESC) into neural 

stem cells (lt-NES®) and their neuronal progeny. MicroRNA-9/9* (miR-9/9*) was 

among the microRNAs that showed a strikingly differential expression pattern. 

It was barely detectable in hESC, whereas it was found expressed in lt-NES® and 

up-regulated in differentiating neurons. 

To gain insight into the potential roles of miR-9/9* in our long-term self-renewing 

lt-NES®, we made use of a lentiviral-based overexpression system and achieved 

overexpression of the mature forms of both miR-9 and miR-9* at comparable 

levels. Despite the presence of growth factors (i.e. FGF2 and EGF) in the culture 

medium, miR-9/9* overexpression induced a reduction in proliferation and an 

increase in differentiation of lt-NES®, as assessed by BrdU incorporation and 

expression of neuronal markers. By exposing lt-NES® to individual synthetic 

mimics of miR-9 and miR-9*, we could demonstrate that the promotion of 

differentiation is due both to miR-9 and miR-9*, whereas the anti-proliferative 

effect is mainly exerted by miR-9* alone. We are currently investigating potential 

connections between miR-9/9* and signaling pathways that are relevant in 

neural stem cell maintenance and differentiation. 

Our experimental approach provides an elegant way to assess the role of 

microRNAs – including bifunctional ones such as miR-9/9* - during early human 

neural development. It further offers a promising tool for the modulation of 

microRNA levels to gain control on the fate of human stem cells.

87 


Retrograde tracing of neurons in the rodents lateral 

bladder wall: an experimental study with fluorescent 

latex beads: mucosal layers of the bladder vs. small 

intestinal mucosa 

Anna Schueth 1,2 , Gommert A. van Koeveringe 1 

1 Maastricht University Medical Centre, azM, Department of Urology, Maastricht, The Netherlands; 

2 Institute of Anatomy, University of Luebeck, Luebeck, Germany. 

In previous studies it was shown that latex beads (LB) of different sizes (20 – 

200 nm), experimentally applied to the intact murine small intestinal mucosa, 

were taken up into the tissue. The LB were detected at least 20 minutes after 

application in different cells lying within the epithelial and subepithelial tissue of 

the mucosa. Both via intravital experiments by means of 2- photon microscopy 

(TPLSM) and confocal microscopy (CLSM) as a reference technique using dissected 

parts of the small intestine, the uptake and localization of LB in the small intestinal 

tissue could be detected and analyzed. Especially, TPLSM was used to investigate 

the particle uptake and further processing in anaesthetized and living mice 

under physiological conditions up to 8 hours. The idea is now, to transfer this 

technique and experimental set-up, initially described with the small intestinal 

mucosa, in an adapted way to mucosal layers of the bladder in rodents. With 

these investigations, mapping of the whole bladder and the neurons (Koebbert 

et al., 2000), as a matter of special interest, starting from the lateral bladder wall 

and ending up in the sacral cord could take place. Unlike the small intestine, 

the bladder has a triple innervation (pudendus and pelvic nerve, going to the 

sacral part and the hypogastric nerve, going to the thoracolumbar spinal cord). 

This could be achieved by applying the fluorescent LB to the rodent’s bladder 

mucosa. A subsequent tracing of the particle routes and their distribution within 

the lateral bladder wall could take place. Immune-histochemical techniques can 

be used in order to further analyse the dissected tissue.

88 


Simultaneous activation of the presynaptic 

cannabinoid CB 1 receptor attenuates the function 

of the presynaptic muscarine M 2 receptor and the δ 

opioid receptor 

Kirsten Schulte, Eberhard Schlicker 

Department of Pharmacology and Toxicology, University of Bonn, 53125 Bonn, Germany. 

We studied whether the presynaptic CB 1 receptor influences the function of other 

types of presynaptic inhibitory receptors. The inhibitory effect of the muscarine 

receptor agonist oxotremorine 10 µM on acetylcholine release was 61±3 and 

78±1 % in hippocampal slices from wild-type (WT) and CB 1 receptor knockout 

(CB 1 KO) mice, respectively (P

89 


Persistent spatial memory improvement after 

phosphodiesterase type 4d inhibition in the APPswe/ 

PS1dE9 mouse model of Alzheimer’s disease 

Annerieke SR Sierksma 1 , Daniel LA van den Hove 1,2 , Olga Bruno 3 , Ernesto Fedele 4 , 

Tim Vanmierlo 1 , Roberta Ricciarelli 4 , Harry WM Steinbusch 1 , Jos Prickaerts 1 

1 Department of Psychiatry and Neuropsychology, School for Mental Health and Neuroscience, Maastricht 

University, Maastricht, The Netherlands; 2 Department of Psychiatry, Psychosomatics and Psychotherapy, 

University of Würzburg, Würzburg, Germany; 3 Department of Pharmaceutical Sciences, University of 

Genoa, Genoa, Italy; 4 Department of Experimental Medicine, University of Genoa, Genoa, Italy. 

Phosphodiesterase type 4 inhibitors (PDE4-Is) have been shown to enhance 

cognition both acutely and chronically. By selectively preventing the breakdown 

of cAMP, it is thought to enhance intracellular signaling and facilitate long-term 

potentiation, the possible neural correlate of memory. Chronic PDE4-I treatment 

with Rolipram of a mouse model of Alzheimer’s disease (AD) has demonstrated a 

persistent improvement on synaptic transmission and spatial memory function. 

However, current PDE4-Is cause severe emesis, making the drug unsuitable for 

human use. GEBR-7b, a recently developed specific PDE4d inhibitor, has been 

shown to have acute cognition-enhancing potential, while lacking the emetic 

side effects. Our aim was to investigate whether chronic GEBR-7b treatment of 

the APPswe/PS1dE9 mouse model of AD could also persistently augment spatial 

memory function. 

For this purpose 5-month-old C57BL/6 (WT) and APP/PS1 mice received a daily 

s.c. injection (5ml/kg) of GEBR-7b (0.001 mg/kg) or vehicle for 21 days. Another 

group received no injections or treatment and were merely handled to control 

for injection stress. After cessation of treatment the animals were exposed to 

the sucrose intake test (SIT), elevated zero maze (EZM), open field (OF), Y-maze, 

object location task (OLT) and the forced swim test (FST), at approximately 7 

months of age. 

OLT results indicated that APP/PS1 mice in general show significantly impaired 

spatial memory performance when compared to WT mice. Moreover, at the 4-hour 

interval, APP/PS1 mice without GEBR-7b treatment were not able to distinguish 

the novel and familiar location, while the APP/PS1 mice treated with GEBR-7b, as 

well all WT mice, were able to discriminate. Interestingly, APP/PS1 mice did show 

higher exploratory behaviour than WT mice in the OLT, which was confirmed by 

greater motility in the OF and EZM. The Y-maze spontaneous alternation test 

showed that there was no difference between groups in spatial working memory 

performance. Although no difference in hedonism or depressive-like behaviour 

could be observed among the groups in the SIT and the FST, respectively. Finally,

90 


APP/PS1 mice showed more anxious behaviour than WT mice as measured by 

the EZM. 

Despite higher object exploration times, APP/PS1 mice showed apparent spatial 

memory deficits compared to WT mice. When chronically treated with GEBR-7b, 

spatial memory performance in APP/PS1 could be restored to WT level, without 

affecting working memory or affective behaviour. Interestingly, the memoryenhancing 

effects of GEBR-7b persisted after cessation of treatment, which 

may be indicative of structural hippocampal changes. Hippocampal analyses are 

underway to provide novel insights into the underlying mechanisms of GEBR-7b 

in preventing spatial memory loss.

91 


The influence of spatial distortion during body 

perception: an eventrelated potential study 

*M. E. Siwek 1,2 , B. Suchan 2 , D. Soria-Bauser 2 , I. Daum 2 . 

1 Cell. and Systemic Neurophysiol., Federal Inst. For Drugs and Med. Devices (BfArM), Bonn, Germany; 

2 Dept. of Neuropsychology, Inst. of Cognitive Neurosci., Bochum, Germany. 

Like human faces, human bodies are indispensable for communication as well 

as for perceiving another’s emotion correctly while providing information about 

gender, age and intentions of other individuals becoming familiar with recurred 

exposure over one’s lifetime. Concerning the high importance both stimuli classes 

for our daily routine, the question arises, what kind of mechanisms are needed 

in the human brain to ensure specialized visual and perceptual processing of 

both, facial and bodily expressions. The current study was aimed to investigate 

behavioral and electrophysiological influences of inversion (rotation around 180°) 

as well as changes of body perception depending upon the degree of spatially 

distorted stimuli referring to a possible disruption of configural processing 

mechanism. Thereby, configural processing continuum distinguishes different 

processing levels of human body shapes according to their spatial relations 

among internal features. 

In this case smoothed body images as one possible stimulus manipulation were 

used in the current study. Spatial distortion or smoothing was done by Gaussian 

filtering (low-pass filtering) of the original pictures with filter widths of 1, 3, 5 

and 7 mm. Thereby, the reduction in high-frequency noise by using low- pass 

filters is obtained at the cost of a loss of spatial resolution. Concerning this the 

presentation of less information/ details containing within a stimuli caused by 

different smoothing degrees might influence the encoding of basic features that 

are necessary for the processing of body parts and individual representation 

of the body as a whole. On the electrophysiological level body specific eventrelated 

components, N170 and P100 were investigated. For both, N170 and P100 

amplitudes, the inversion effect was replicated. On the behavioral level increasing 

smoothing steps lead to higher error rates and efficiency scores. In case of 

electrophysiological measures an effect of spatial distortion was obtained for the 

P100 latency but not for the N170 component. Overall, the findings support that 

the N170 being elicited by bodies is a robust phenomena concerning spatially 

manipulated depictions of human bodies.

92 


Migration of microglia in the embryonic neocortex 

Sophie Smolders 1 , Nina Swinnen 1 , Bert Brône 1 , Jean-Michel Rigo 1 

1 Hasselt University, BIOMED, Physiology group, Diepenbeek, Belgium. 

Microglia, the macrophages of the central nervous system (CNS), are assumed 

to support the architecture and functional maturation of the developing CNS, 

through secretion of various factors. After invading the quail CNS early during 

embryogenesis, retinal microglia migrate in tangential and radial directions 

using Müller cells (retinal radial glia). Preliminary results of our laboratory show 

contact between microglia and radial cells during murine neocorticogenesis. 

Moreover, in vitro, microglia express receptors for extracellular matrix proteins 

present in the developing brain. Therefore, this study aims to elucidate the 

migratory behavior of microglia and to determine the presence of adhesion 

molecules on microglia and radial cells, which could allow molecular interaction 

during neocorticogenesis. Immunohistochemical stainings were performed on 

fixed brain tissue of C57BL/6 CX3CR1 +/eGFP mice embryos (Embryonic day (E) 12.5- 

15.5). Cortical microglial location and protrusion morphology were analyzed 

using a home-made matlab plug-in. Microglial migration was recorded in CX3CR1 

+/eGFP acute brain slices using live imaging and analyzed using MTrackJ in ImageJ. 

Statistical analyses were applied in GraphPad Prism with P

93 


Cloning and production of anti inflammatory 

antibodies against A-Beta 

Jo Stevens, Pilar Martinez, Marc De Baets, Harry Steinbusch, Mario Losen 

Maastricht University, School of Mental Health and Neuroscience, Universiteitssingel 50, 6229 ER, Maastricht, 

The Netherlands. 

Active and passive immunization studies targeting different forms of Abeta 

in Alzheimer’s disease have demonstrated antibody-mediated reduction of 

amyloid plaque load. However, these treatments may lead to T-cell-mediated 

meningoencephalitis and inflammation. Since antibodies can exert inflammatory 

responses, we propose that the effector mechanisms of therapeutic antibodies 

need to be carefully controlled: we aim to study the contribution of these 

mechanisms to the therapeutic effect (plaque clearing and improvement of 

memory function) and possible side effects using pro-inflammatory mouse IgG2a 

and anti-inflammatory mouse IgG1 antibodies in the 5xFAD transgenic AD mouse 

model with plaque formation starting at 2 months of age. Here we show the 

first results of the production and characterization of 3 clinically relevant antiinflammatory 

antibodies.

94 


γ-secretase dependent phagocytosis of Amyloid-beta 

(Aβ) in microglial cells 

Sonia Tosheva, Jochen Walter 

Department of Neurology, University of Bonn. 

Introduction 

Alzheimer’s disease (AD) is the most common form of dementia characterized 

by key features that include neurofibrillary tangels formation and depositions 

of fibrillar amyloid β-peptides (Aβ) which form senile plaques in the brain. Aβ 

is produced after amyloid precursor protein proteolytic cleavage via β and 

γ-secretase pathways. γ-secretase cleaves many type I proteins thereby affecting 

the protein stability and trafficking, cognitive functions, intracellular signaling 

and calcium homeostasis. This enzyme is a complex of 4 individual proteins but 

the catalytic role is due to Presenilins (PS). And missense mutations in the PS 

genes are a major cause of early onset of familial AD (FAD). 

To address the critical role of γ-secretase in amyloid plaque clearance, we focused 

our work on studying the involvement of microglial cells in this process. The 

microglial phagocytosis of Aβ can be triggered via several receptors like the Low 

density lipoprotein receptor related protein1 (LRP1). LRP1 is a type I membrane 

protein and a γ-secretase substrate. It interacts with APP and facilitates its trafficking 

and processing. LRP1 uses Aβ as a ligand and controls its uptake and transport to 

the lysosomes. There Aβ is degraded by proteolitic enzymes mainly Cathepsin D. 

Methods 

We chose different approaches to study the γ-secretase functions in microglial 

cell lines- BV2 and ES derived microglia (EsDM). Genetic mutations of the 

Presenilin (PS) gene and pharmacological component (DAPT) were used to inhibit 

the γ-secretase activity. 

Results 

Our results showed that γ-secretase is involved in Aβ uptake and decrease in this 

process was seen after γ-secretase inhibition. We reported that Aβ`s receptor 

LRP1 is expressed in microglial cells where its processing and endocytosis is 

γ-secretase dependent. Cells with inhibited γ-secretase activity demonstrated 

impaired LRP1 endocytosis, and Aβ cellular uptake. This also correlated with a 

decrease in degradation and changes in the lysosomal structure and localization. 

Opposite effects were shown in microglia cells overexpressing PS1 where the rate 

of Aβ uptake and its degradation was significantly high.

95 


Conclusion 

Our results indicate that γ-secretase inhibition contribute to Aβ accumulation 

in the brain. Modulation of γ-secretase activity could be used as a promising 

therapeutic target.

96 


Inhibition of WNT signaling impairs growth of 

synovial sarcoma cells 

M. Trautmann 1 , E. Sievers 1 , D. Kindler 1 , A. Koch 3 , R. Büttner 2 , W. Hartmann 1 

1 Institute of Pathology, University Hospital Bonn, Sigmund-Freud Str. 25, 53127 Bonn, Germany; 2 Institute 

of Pathology, University Hospital Cologne, Kerpener Str. 62, 50924 Cologne, Germany; 3 Department 

of Neuropathology, Charité Universitätsmedizin, Charitéplatz 1,10117 Berlin, Germany. 

Synovial sarcoma is a rare malignant soft tissue tumor affecting mainly 

adolescents and young adults. The hallmark of synovial sarcoma is the presence 

of a reciprocal balanced t(X;18) translocation, leading to the fusion of the SS18 

gene to either the SSX1, SSX2 or rarely the SSX4 gene, resulting in a chimeric 

transcriptional modifier. Therapeutic outcome of synovial sarcomas is primarily 

determined by the efficiency of surgery as a high tendency for local relapse is 

documented. Standardized chemo-and radiotherapy are further therapeutic 

options, however, specific targeted therapies are currently not available. 

Recently, several expression profiling studies in mesenchymal malignancies 

revealed gene expression signatures indicating WNT signaling activation in 

synovial sarcomas. 

This study was performed to examine the functional relevance of WNT signaling 

in synovial sarcomas and to evaluate if interference with the WNT signaling 

pathway might represent an option in the development of novel and highly 

selective drugs in the treatment of synovial sarcoma. 

To assess the prevalence of WNT signaling activation in a set of 30 synovial 

sarcoma tumor samples, nuclear staining of β-catenin was analyzed 

immunohistochemically. Nuclear β-catenin signals were observed in a significant 

subset of these tumors, indicating activation of the WNT signaling pathway. 

In order to evaluate whether WNT activation is molecularly dependent on 

the SS18/SSX fusion proteins, tetracycline-inducible systems overexpressing 

the SS18/SSX fusion proteins were established in T-Rex293 cells. In luciferase 

reporter assays employing the TOP-/FOPflash system, expression of SS18/SSX 

proteins effectively activated TCF/β-catenin mediated transcriptional activity, 

which was associated with nuclear recruitment of β-catenin. Five human synovial 

sarcoma cell lines were subsequently treated with small molecular inhibitors of 

WNT signaling. In MTT assays, a significant dose-dependent inhibition of cellular 

growth were observed, which was accompanied by decreased expression of the 

WNT downstream targets c-Myc and Cyclin D1. In flow cytometric analyses, the 

growth effects exerted by the inhibitors were shown to be due to a reduction of 

cellular proliferation combined with an increase of apoptosis. 

In summary, our data emphasize the pivotal role of WNT signaling in synovial

97 


sarcoma and indicate its functional dependence on the characteristic SS18/SSX 

translocation. Furthermore, our study demonstrates that targeting the WNT 

signaling pathway provides a specific, molecularly founded therapeutic strategy 

in the treatment of synovial sarcoma. Additional functional studies in vitro and 

in vivo are required to further understand the role of WNT signaling and its 

therapeutic applicability in synovial sarcomas.

98 


Aberrant modular organization of cerebral 

functional networks in cognitive impaired children 

with frontal lobe epilepsy 

Vaessen M. 1,2,4 , Heerink J. 1 , Braakman H. 2,3,4 , Hofman P. 2,4 , Aldenkamp A. 2,3,4 , 

Jansen J. 1,4 , Backes W. 1,4 

1 Department of Radiology, Maastricht University Medical Centre, Maastricht, The Netherlands; 2 Department 

of Research and Development, Epilepsy Centre Kempenhaeghe, Heeze, The Netherlands; 3 Department 

of Neurology, Maastricht University Medical Centre, Maastricht, The Netherlands; 4 Research 

School for Mental Health and Neuroscience, Maastricht University Medical Centre, Maastricht, The 

Netherlands. 

Introduction 

Many children suffering from frontal lobe epilepsy (FLE) have significant cognitive 

impairments, which may hinder their scholarly development. The underlying 

mechanism or neuronal correlate of this cognitive co-morbidity has not yet been 

unraveled. Using resting-state functional magnetic resonance imaging (RS-fMRI), 

a technique that enables the measurement of intrinsic functional connections in 

the brain, we investigated whether a quantitative analysis of cerebral network 

characteristics might be associated with epilepsy and co-morbid cognitive 

problems. 

Methods 

We included 37 children with FLE, compared them with 41 healthy age-matched 

controls, and determined their cognitive performance by means of a cognitive 

visual searching task. A connectivity matrix was generated for each subject by 

calculating the correlation of the time-signals of 82 cortical and sub-cortical brain 

regions. From this connectivity matrix network parameters were calculated. 

Results 

In children with FLE, decreasing cognitive performance was accompanied by 

higher modularity scores and a different modular organization of the brain. 

Conclusions 

Cognitively impaired patients showed highest modularity scores, implying 

that the constituting sub-networks are less mutually connected. Therefore, it 

is concluded they have a more profound deterioration in the organization of 

functional networks. We suggest that cognitive problems might be partially 

related to this abnormal large-scale functional connectivity of the brain.

99 


Brain-derived neurotrophic factor (BDNF), a bridge 

between depression and Alzheimer’s disease 

Tim Vanmierlo, Jochen De Vry, Caroline Hammels, Annerieke Sierksma, Denise 

Hermes, Harry Steinbusch and Jos Prickaerts. 

School for Mental Health and Neuroscience, Maastricht University, Maastricht, The Netherlands 

There is ample evidence suggesting that a history of major depression constitutes a 

risk factor for the development of Alzheimer’s disease (AD) later in life. Decreased 

levels of the growth factor brain-derived neurotrophic factor (BDNF) play a key 

role in the reduced plasticity in the hippocampus of patients with depression as 

well as in patients with AD. We hypothesize that hippocampal overexpression 

of tropomyosine receptor kinase (TrkB), the high affinity receptor of BDNF, will 

attenuate and delay the onset of AD pathology. To test this hypothesis we used 

the APPswe/PS1dE9 mouse model of AD. Hippocampal TrkB overexpression 

was established via a current controlled stereotactic microelectroporation of a 

plasmid stably expressing TrkB at the start of AD pathology, i.e. plaques formation, 

at 6 months of age. Memory was scored at 7 months of age, when cognitive 

decline is expected to start, in the object location task, the Morris water escape 

maze and the spatial alteration Y-maze. In addition, depression and anxiety 

related behavior were respectively assessed in the sucrose preference test, the 

forced swim task and the elevated zero maze. Electroporation efficiency will be 

validated by immunohistochemistry (IHC). Manifestation of the AD pathology 

on the molecular level will be quantified by ELISA and IHC. First results showed 

surprisingly that TrkB overexpression in the hippocampus impaired object location 

memory in our AD mice. In contrast, depression–like behavior in the forced swim 

test was attenuated in these AD mice. No effects of TrkB overexpression were 

observed in wild-type control mice. Biochemical and IHC analyses are underway 

to further investigate this possible TrkB-mediated distinction in cognitive and 

affective behavior in AD mice.

100 


Ceramide signaling regulates the effects of fetal 

asphyctic preconditioning 

Evi Vlassaks 1,2 , Eveline Strackx 1,2 , Chiara Mencarelli 2 , Hans Vles 3 , Marc de Baets 2 , 

Danilo Gavilanes 1 , Pilar Martinez 2 

1 Department of Pediatrics – Division of Neonatology, Maastricht University Medical Center; 2 Department 

of Neuropsychology – Division Neuroscience, Maastricht University; 3 Child Neurology, Maastricht 

University Medical Center. 

Introduction 

Nowadays, only few effective treatments are available for perinatal asphyxia. 

One promising approach in the search of new therapies is investigating the 

mechanisms underlying asphyctic preconditioning. Ceramide signaling is one of 

the stress responses participating in various biological processes, including the 

development of post-asphyctic injury. Moreover, it has been shown that ceramide 

exerts protective effects against hypoxia/ischemia. Hence, an understanding of 

the ceramide signaling in asphyctic preconditioning might provide new insights 

into the development of new therapeutics. 

Materials & methods 

A mild asphyctic preconditioning stimulus was induced at E17 by completely 

clamping both the uterine and the ovarian circulation for 30 min. At birth, severe 

perinatal asphyxia was induced by submersing the uterine horns in a saline bad 

for 19 min. Pups were sacrificed at different time points after both insults. The 

effects of preconditioning and asphyxia on the ceramide pathway were assessed 

by investigating relevant enzymes and transporters in both brain and liver by 

quantitative RT-PCR. 

Results 

In the prenatal brain, we found increased mRNA levels of the enzymes Lass1 

and SMS and increased levels of the transporters GPBP and CERT after the 

preconditioning stimulus compared to controls. These central effects on ceramide 

signaling were preceded by peripheral mRNA changes as showed by increased 

hepatic mRNA levels of Lass1, SMS, GPBP and CERT. After birth no changes were 

observed anymore. 

Conclusion 

Fetal asphyctic preconditioning influences the ceramide pathway by increasing 

ceramide synthesis and transport. Accordingly, ceramides might play a role in the 

induction of tolerance to asphyctic insults. Deeper knowledge of the involved cell

101 


signaling pathways will help in understanding the preconditioning phenomenon, 

which subsequently opens opportunities for the development of new therapies.

102 


Memory deficits in transgenic Huntington’s disease 

rats 

Dagmar Zeef 1,2,3 , Nick van Goethem 1,3 , Rinske Vlamings 1,2,3 , Jos Prickaerts 1,3 , 

Yasin Temel 1,2,3 

Departments of 1 Neuroscience and 2 Neurosurgery, Maastricht University Medical School, Maastricht, 

The Netherlands; 3 European Graduate School of Neuroscience (EURON). 

Besides motor abnormalities, Huntington’s Disease (HD) patients suffer from 

progressive cognitive impairments. Recently, a new transgenic rat model for HD 

has been introduced, which is currently being characterized in detail. Here, we 

investigated the memory functions of 10- and 16-months old tgHD rats and 16 

months old wild-type (WT) littermates in the object location (OLT) and the object 

recognition tests (ORT) representing, respectively, visuospatial and visual object 

memory. We found that the differences in exploration times between the familiar 

and the novel object, in the ORT, were significant in WT rats, but not in the tgHD 

rats at the age of 10- and 16-months. Post-hoc analysis showed that the WT rats 

had a significantly better object recognition performance when compared to 

the tgHD rats at the age of 16 months. In the OLT, the difference in exploration 

times between the familiar and the novel location reached significance in the 

WT animals, but no differences in both the 10 and the 16-months old tgHD rats 

were found. Additionally, post-hoc analysis revealed that the 16-months old 

tgHD rats had a significantly less location recognition performance compared 

to the WT animals. The lack of preference for the novel object or for the novel 

place indicates that the tgHD rats have impaired visuospatial and visual object 

memory. This is present already at the early stage of the disease and progresses 

with ageing. These memory deficits and their progressive nature mimic some 

aspects of cognitive symptoms of HD patients and seem to make this tgHD model 

relevant for interventional studies. 

This study was funded by the Cure Huntington’s disease initiative.

List of Participants 

103 


104 


Nuersailike Abuduwali 


PhD-Student 

serk_omar@yahoo.com 

Sven Akkerman 


PhD student 

s.akkerman@maastrichtuniversity.nl 

Christina Albus 



chralbus@uni-bonn.de 

Jana Anschlag 



JanaA@gmx.de 

Stefan Aretz, PD Dr. 


staretz@uni-bonn.de 

Ariel Avila 

Hasselt University 


ariel.avilamacaya@uhasselt.be 

Buket Basmanav 



basmanav@uni-bonn.de 

Heinz Beck, Prof. Dr. 


Heinz.Beck@ukb.uni-bonn.de 

105 


Jessica Becker 



jbecker@uni-bonn.de 

René Besseling 



r.m.h.besseling@gmail.com 

Clara Beutner 



cbeutner@uni-bonn.de 

Sandra Blaess, Dr. 


sandra.blaess@uni-bonn.de 

Andreas Bock 



a.bock@uni-bonn.de 

Gabriela Bodea 



gbodea@uni-bonn.de 

Liviu Bodea 



lbodea@uni-bonn.de 

Eva Bollen 



e.bollen@maastrichtuniversity.nl

Iris Bomilcar-Focke 


Master student 

irisbomilcar@googlemail.com 

Lodovica Borghese, Dr. 


borghese@uni-bonn.de 

Verena Borm 



vborm@uni-bonn.de 

Oliver Braganza 



olli_tb@yahoo.com 

Gary Brook, Dr. 

RWTH Aachen 

gbrook@ukaachen.de 

Cin-He Chang 



cin-he.chang@ukb.uni-bonn.de 

Thierry Charlier, Dr. 

University of Liege 

tcharlier@ulg.ac.be 

Sven Cichon, Prof. Dr. 


scichon@uni-bonn.de 

Claudia Cornelissen 



ccorneli@uni-bonn.de 

106 


Kimberly Cox 



kimberly.cox@maastrichtuniversity.nl 

Holger Dannenberg 



hdannenb@uni-bonn.de 

Marc De Baets, Prof. Dr. 


m.debaets@maastrichtuniversity.nl 

Marie Decock 

Université Catholique de Louvain 


marie.decock@uclouvain.be 

Frank Dennissen 



f.dennissen@maastrichtuniversity.nl 

Ilse Dewachter, Dr. 


ilse.dewachter@uclouvain.be 

Kristina Dobrindt 



Kristina.Dobrindt@uni-bonn.de 

Matthias Eckhardt, PD Dr. 


eckhardt@uni-bonn.de

Marianne Eisenhardt 



Marianne.Eisenhardt@ukb.unibonn.de 

Daniela Evers 



d.evers@uni-bonn.de 

Bernd Evert, PD Dr. 


b.evert@uni-bonn.de 

Bernd Fleischmann, Prof. Dr. 


bfleisch@uni-bonn.de 

Stephanie Friedrichs 



sfriedri@uni-bonn.de 

Mary Gazea 



m.gazea@googlemail.com 

Jose Luis Gerardo Nava 

Maastricht University / 

RWTH Aachen University 


jgerardonava@ukaachen.de 

Romina Gentier 



r.gentier@maastrichtuniversity.nl 

107 


Debora Gerlach 



DeboraGerlach@gmx.de 

Volkmar Gieselmann, Prof. Dr. 


vgieselm@uni-bonn.de 

Andreas Glässner 



Andreas.Glaessner@ukb.uni-bonn.de 

Alejandro Gomez 



a.gomez@maastrichtuniversity.nl 

Michaela Granzow 



Michaela.Granzow@ukb.uni-bonn.de 

Stephanie Griemsmann 



Stephanie.Griemsmann@ukb.unibonn.de 

Caroline Hammels 



c.hammels@maastrichtuniversity.nl 

Robert Hardt 



hardt@uni-bonn.de

Wolfgang Hartmann, PD Dr. 


wolfgang.hartmann@uk-koeln.de 

Stefanie Heilmann 



sheilman@uni-bonn.de 

Aurélie Hendrickx 



aurelie.hendrickx@uclouvain.be 

Verena Herl 



Verena.Herl@ukb.uni-bonn.de 

Stefan Herms 


stefan.herms@uni-bonn.de 

Dorothee Hodde 

RWTH Aachen University, Germany 


dhodde@ukaachen.de 

Per Hoffmann, Dr. 


Per.Hoffmann@uni-bonn.de 

Ramona Hohnen 



r.hohnen@student. 

maastrichtuniversity.nl 

108 


Sukanya Horpoapan 



s6suhorp@uni-bonn.de 

Ronald Jabs, PD Dr. 


Ronald.Jabs@ukb.uni-bonn.de 

Ali Jahanshahi 



a.jahanshahianvar@ 


Anna Kabanova 



anna.kabanova@uni-bonn.de 

Pascal Kienlen-Campard, Prof. Dr. 


pascal.kienlen-campard@uclouvain.be 

Carsten Kilgus 



ckilgus@uni-bonn.de 

Frank Kirchhoff, Prof. Dr. 

University of Saarland 

frank.kirchhoff@uks.eu 

Anna-Lena Klauke 



anna-lena.klauke@uni-bonn.de

Sabine Klein 



Sabine.Klein@ukb.uni-bonn.de 

Jens Kopatz 



jens.kopatz@uni-bonn.de 

Andreas Kriebel 

RWTH Aachen University, Germany 


andreas.kriebel@rwth-aachen.de 

Pierre Leprince, Dr. 


pleprince@ulg.ac.be 

Andreas Lindstrot 



andreas.lindstrot@ukb.uni-bonn.de 

Bettina Linnartz, Dr. 


linnartz@uni-bonn.de 

Julia Lodder-Gadaczek 



julia.lodder@uni-bonn.de 

Mario Losen, Dr. 


m.losen@maastrichtuniversity.nl 

109 


Anurag Maheshwari 



anurag.maheshwari@uhasselt.be 

E. Mandelkow, Prof. Dr. 

MPI Hamburg/German Center for 

Neurodegenerative Diseases Bonn 

mand@mpasmb.desy.de 

Claudia Marinangeli 



claudia.marinangeli@uclouvain.be 

Pilar Martinez, Dr. 


p.martinez@maastrichtuniversity.nl 

Dix Meiberth 



dixurs@gmx.de 

Irene Melo de Carvalho 



irene.melo@gmail.com 

Chiara Mencarelli 



c.mencarelli@maastrichtuniversity.nl 

Roopika Menon 



mroopika@gmail.com

Jörg Mey, Dr. 


jmey@sescam.jccm.es 

Katrin Michel 



michel.katrin@yahoo.com 

Kerstin Morcinek 



kerstin.morcinek@uk-koeln.de 

Laura Mürtz 



lauramue@uni-bonn.de 

Rohit Nalavade 



Rohit.Nalavade@dzne.de 

Kim Neitzert 



kim.neitzert@uni-bonn.de 

Tien Thanh Nguyen 



Thanh_Tien.Nguyen@ukb.uni-bonn.de 

Neville-Andrew Niessen 



NA.Niessen@ulg.ac.be 

110 


Marjan Nokhbesaim 



m.saim@uni-bonn.de 

Vanessa Nunes de Paiva 



nunesp@uni-bonn.de 

Astrid Ooms 



astrid.ooms@uni-bonn.de 

Annika Ottersbach 



annika85@uni-bonn.de 

Geke Overvliet 



geke_overvliet@hotmail.com 

Milan Pabst 



milanpabst@googlemail.com 

Stefan Passlick 



Stefan.Passlick@ukb.uni-bonn.de 

Jody Pawluski, Dr. 


j.pawluski@maastrichtuniversity.nl

Viktoriya Peeva 



Viktoriya.Peeva@ukb.uni-bonn.de 

Leonie Pothmann 



LeoniePothmann@web.de 

Jos Prickaerts, Dr. 


jos.prickaerts@maastrichtuniversity.nl 

Lutz Priebe 



lpriebe@uni-bonn.de 

Ine Rayen 



i.rayen@maastrichtuniversity.nl 

Stefan Remy, Dr. 


Stefan.Remy@dzne.de 

Sarah Rieck 



srieck@uni-bonn.de 

Jean-Michel Rigo, Prof. Dr. 

University of Hasselt 

jeanmichel.rigo@uhasselt.be 

111 


Beate Roese-Koerner 



beate.syttkus@uni-bonn.de 

Karin Rohleder 



karinrohleder@hotmail.de 

Andrea Rottländer 



andrearottlaender@googlemail.com 

Hannsjörg Schröder, Prof. Dr. 


schroeder.anatomie@uni-koeln.de 

Anna Schueth 



anna.schueth@maastrichtuniversity.nl 

Kirsten Schulte 



kirsten.schulte@uni-bonn.de 

Nicole Senden, Dr. 


n.senden@maastrichtuniversity.nl 

Anahita Sharaz 



shahraz@uni-bonn.de

Florian Siedek 



siedek@uni-bonn.de 

Annerieke Sierksma 



a.sierksma@maastrichtuniversity.nl 

Magdalena Elisabeth Siwek 

BfArM, Bonn 


magdalena.Siwek@bfarm.de 

Sophie Smolders 



sophie.smolders@student.uhasselt.be 

Michelle Sparnaaij 



m.sparnaaij@maastrichtuniversity.nl 

Cosmin Stancu 



cosminstancu@gmail.com 

Harry Steinbusch, Prof. Dr. 


h.steinbusch@maastrichtuniversity.nl 

Christian Steinhäuser, Prof. Dr. 


Christian.Steinhaeuser@ukb.unibonn.de 

112 


Jo Stevens 



jo.stevens@maastrichtuniversity.nl 

Christiane Stieber, Dr. 


cstieber@uni-bonn.de 

Moritz Tacke 



Svenja Ternes 



sternes@uni-bonn.de 

Martin Theis, Dr. 


Martin.Theis@ukb.uni-bonn.de 

Sonia Tosheva 



bineval_nondescript3@yahoo.com 

Marcel Trautmann 



trautmann@uni-bonn.de 

Iris Ullrich, 


theme@uni-bonn.de 

Maarten Vaessen 



mvaessen@gmail.com

Sylvie van der Kruijs 



KruijsS@kempenhaeghe.nl 

Fred van Leeuwen, Dr. 


f.vanleeuwen@maastrichtuniversity.nl 

Tim Vanmierlo, Dr. 


t.vanmierlo@maastrichtuniversity.nl 

Hristo Varbanov 



hvarbano@smail.uni-koeln.de 

Evi Vlassaks 



e.vlassaks@maastrichtuniversity.nl 

Sarah Vosen 



svosen@uni-bonn.de 

Jochen Walter, Prof. Dr. 


Jochen.Walter@ukb.uni-bonn.de 

Andrea Weber, Dr. 


aweber@ibmb.uni-bonn.de 

Thérëse Wolfenstetter 



twolfenstetter@aol.com 

113 


Youssef Yakkioui 



y.yakkioui@student. 


Dagmar Helena Zeef 



dh.zeef@Maastrichtuniversity.nl 

Jiong Zhang 



Jiong.Zhang@ukb.uni-bonn.de

Director of Euron 


Coordinating Office 

Nicole Senden [Program Coordinator] 

Peggy Bisschoff [Communication Officer/Webmaster] 

Marie-Thérèse Moers [Secretary] 

European Graduate School of Neuroscience 

[Euron] 


School for Mental Health and Neuroscience [MHeNS] 

Faculty of Health, Medicine and Life Sciences 

Universiteitssingel 50 

6229 ER Maastricht [NL] 

T +31[43] 388 41 30 / 388 10 21 

F +31[43] 367 10 96 

E info@euronschool.eu 

For more information contact our Office or visit 

the website at: 

www.euronschool.eu 

www.neuronworkshops.nl 

www.maastrichtuniversity.nl/mhens

EURON and THEME joint PhD meeting

Create successful ePaper yourself

Delete template?

Save as template?