Book of abstracts - British Neuroscience Association

57.13
Characterization of a focal eae-mog model for the study of
reactivation of CNS lesions by a systemic inflammatory response
Serres S 1, Jiang Y 2, Amor S 3, Anthony D C 2, Sibson N R 1
1Experimental Neuroimaging Group, Department of Physiology,
Anatomy & Genetics, and, 2Experimental Neuropathology Lab.,
Department of Pharmacology, University of Oxford, UK, 3Department
of Immunobiology, Biomedical Primate Research Centre, Rijswijk, The
Netherlands,
Multiple sclerosis (MS) is a chronic inflammatory disorder which can
display a relapsing-remitting form of disease. It is thought that
bacterial and viral infections may trigger relapses and progression of
MS. Our aim was to characterise a focal EAE-MOG model targeted in
the brain by intracerebral injection of cytokines, and to determine
whether this lesion could be reactivated by a systemic inflammatory
response.
MOG model was induced in Male Lewis rats by injection of MOG
peptide (35-55) or MOG protein following three weeks later by
injection of cytokines in the left hemisphere. Four weeks later, the
animals were challenged by i.p injection of lipoplysaccharide (LPS;
100 μg/kg). We caracterized the lesion by both MRI and
Immunohistochemistry.
T2-weighted images shows a disruption of the signal in the injection
site (corpus callosum). The longitudinal study reveals the presence of
a chronic hypointense signal in the striatum below the injection site,
the increase in ventricle size and incomplete recovery of the signal
from the corpus callosum.
The immunostaining correlated with the hypointense signal and the
disruption of the corpus callosum structure, suggesting involvement of
both activated macrophage and microglia.
Induction of a systemmic inflammatory response with LPS challenge,
resulted in an increase in the left/right rCBV ratio with recruitment of
macrophages as revealed by ED1 staining. These findings show that
the reactivation of inflammatory processes within a previously
quiescent CNS lesion can be obtained after a systemmic inflammatory
response to a bacterial endotoxin in a clinically-relevent model of MS.
57.14
Localisation of the PARK8 protein kinase LRRK2 in cellular models
and human tissue
Sancho R M, Kingsbury A E*, Bandopadhyay R*, Harvey R J, Harvey K
Department of Pharmacology, The School of Pharmacy, 29-39 Brunswick
Square, London WC1N 1AX; *Reta Lila Weston, Institute, University
College London, London WC1N 1PJ
Missense mutations in the protein kinase LRRK2 are the most common
known cause of Parkinson’s disease (PD), found in ~ 5% and 1.6% of
patients with familial and idiopathic PD, respectively. LRRK2 contains
leucine-rich repeats and GTPase, COR (C-terminal of Roc), kinase and
WD40 domains. Mutations are found in all of these domains, but it is
unclear how these changes cause PD. To enable us to investigate LRRK2
expression, we raised two polyclonal antibodies (pAbs) against LRRK2. In
HEK cells, endogenous and exogenous LRRK2 (encoded by various
mammalian expression constructs) was clearly recognised by antibody
BC300-267. In Western blots from human brain lysates BC300-267
recognised a specific band of ~260 kDa that was completely abolished in
competition experiments using an excess of the peptide used for
immunization. BC300-267 was further tested on post-mortem tissue from
neurologically normal and sporadic PD cases, revealing diffuse staining of
cell bodies and processes of midbrain melanized neurons and selected
brainstem and midbrain nuclei. In nine separate pathologically-verified
sporadic PD cases, LRRK2 immunoreactivity was detected in the inner part
of the halo of cerebellar and midbrain LBs, with some variation in staining
intensity between individual LBs. Interestingly, no LRRK2 immunopositive
cortical LBs were seen in these cases. Lastly, using the yeast two-hybrid
system and mammalian cellular models, we demonstrate that LRRK2
interacts with synphilin-1. Confocal imaging showed colocalisation of
LRRK2 and synphilin-1 in the cytoplasm and perinuclear aggregates with a
central core of synphilin-1 and an LRRK2 halo.
57.15
Anosmia in subjects with dementia is associated with Lewy
pathology in the cortical olfactory pathway.
Hubbard P, Esiri M M, Smith A D, King E, Reading M, Nagy Z,
McShane R
1,6 University of Birmingham, Birmingham, UK., 2,5 Neuropath. Dept,
John Radcliffe Hospital, Oxford, UK, 3, Oxford Centre for Gene
Function University of Oxford,Oxford, UK, 4, OPTIMA Radcliffe
Infirmary NHS Trust Oxford, UK , 7,Dept Old Age Dept Dept Old Age
Psychiatry, Churchill Hospital, Oxford,UK
Neurodegenerative diseases currently affect over 750,000 people in
the UK. The most common disorders leading to dementia are
Alzheimer’s disease and dementia with Lewy bodies. So far the
definitive diagnosis of these dementias can only be confirmed postmortem.
However, loss of smell (anosmia) is an early symptom in
patients who develop dementia. The use of a smell test has been
proposed as an early diagnostic procedure and distinguishes those
with early dementia with Lewy bodies (DLB) from those with early
Alzheimer’s disease.
The present study aimed to examine the relationship between
anosmia and the presence of Lewy body pathology in the olfactory
pathways.
Participants in the Oxford Project to Investigate Memory and Ageing
were tested for basic olfactory function during life. Full ethical
permission was obtained for use of tissue. Tissue was taken from 5
areas of the olfactory pathway; the Olfactory Tract/Bulb, the insertion
of the Olfactory Tract, the orbito-frontal cortex, the hippocampus and
the amygdala. Lewy bodies were detected using
immunohistochemistry to alpha-synuclein.
Results show that the quantity of Lewy Body pathology in the olfactory
system varied, but may follow a particular pattern of development. The
presence of pathology in the cortical pathway but not the limbic
pathway is associated with anosmia, and is further evidence towards
the use of a smell test to aid diagnosis of neurodegenerative diseases.
57.16
A fibroblast growth factor receptor 1 agonist reduces the loss of
hippocampal dendritic spines caused by beta-amyloid(25-35)
Corbett N J, Stewart M G, Gabbott P L, Klementiev B, Davies H A, Colyer F
M, Berezin V, Bock E
The Open University, , Department of Biological Sciences, Walton Hall,
Milton Keynes, Buckinghamshire, MK7 6AA,
In Alzheimer’s disease (AD), β-amyloid (Aβ) affects neural circuits
underlying learning and memory. Fibroblast Growth Loop (FGL), a
pentadecapeptide which mimicks the heterophilic binding of neural cell
adhesion molecule (NCAM) to Fibroblast Growth Factor Receptor 1
(FGFR1), has the potential to significantly reduce the neurodegeneration
seen in AD.
Aβ25-35 was injected intracerebroventricularly (icv) into the right
hemisphere of 2 groups of rats (5mg/15μl; n=6/group). Eight days after Aβ
injection, animals received either a subcutaneous injection of FGL
(10.8mg/kg) or distilled water (dw) every 3 days (Aβ-FGL, Aβ–dw groups
respectively). Another group (dw-FGL: n=6) had dw injected icv followed by
FGL treatment whilst the control group (dw-dw: n=6) received both forms of
dw.
On day 25, animals were transcardially perfused with 2%
paraformaldehyde and 3.75% acrolein in 0.1M phosphate buffer (pH 7.4).
Using the ‘Single-section’ Golgi method on 100μm thick coronal sections,
the density of dendritic-spines along CA1 pyramidal apical dendrites in the
stratum radiatum were obtained (Neurolucida). This area of spiny dendrites
was chosen due to the excitatory interactions with the Schaffer axon
collaterals that are notably damaged in AD.
Results indicated that the Aβ-dw group had a significantly lower average
spine density at 1.5 ± 0.1μm-1 (P