identification of neuroprotective genes against alpha - acumen - The ...

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µl Microliter miRNA MicroRNA mg Milligram ml Milliliter mM Millimolar MAPK Mitogen-activated protein kinase MAPKK Mitogen-activated protein kinase kinase MAPKKK Mitogen-activated protein kinase kinase kinase n/a Not applicable NGM Nematode growth medium PARK Parkinson disease gene PCR Polymerase chain reaction PD Parkinson disease PDE Posterior deirid neuron RING Really interesting new gene RNA Ribonucleic acid RNAi RNA interference Roc Ras <strong>of</strong> complex ROS Reactive oxygen species rpm Revolutions per minute RT Room temperature (25 °C) v
RT-PCR Reverse transcriptase polymerase chain reaction SAGE Serial analysis <strong>of</strong> gene expression SD Standard deviation SDS-PAGE Sodium dodecyl sulfate polyacrylamide gel electrophoresis SNP Single nucleotide polymorphism UPR Unfolded protein response UPS Ubiquitin-proteasome system UTR Untranslated region C. elegans Proteins AGE-1 Aging alteration 1 (phosphoinositide 3-kinase) ATGR-7 Autophagy 7 BAR-1 Beta-catenin/armadillo related 1 CDK-5 Cyclin dependent kinase 5 CED-3 Cell death abnormality 3 (caspase) CLK-1 Clock 1 (demethoxyubiquinone hydroxylase) CMK-1 Calcium/calmodulin-dependent protein kinase 1 CSNK-1 Casein kinase 1 DAF-2 Abnormal dauer formation 2 (insulin receptor) DAF-16 Abnormal dauer formation 16 (forkhead Box 01A) DAT-1 Dopamine transporter 1 vi
Page 1 and 2: IDENTIFICATION OF NEUROPROTECTIVE G
Page 3 and 4: ABSTRACT Recent functional analyses
Page 5: E3 Ubiquitin ligase ER Endoplasmic
Page 9 and 10: TAG-278 Temporary assigned gene 278
Page 11 and 12: p38 Mitogen-activated protein kinas
Page 13 and 14: ACKNOWLEDGMENTS First and foremost,
Page 15 and 16: CONTENTS ABSTRACT .................
Page 17 and 18: a. Abstract .......................
Page 19 and 20: LIST OF FIGURES 1.1. Schematic repr
Page 21 and 22: Parkinson disease CHAPTER ONE INTRO
Page 23 and 24: al., 2006) and torsinA (Cao et al.,
Page 25 and 26: endoplasmic reticulum (ER) to Golgi
Page 27 and 28: c-Jun N-terminal kinase and caspase
Page 29 and 30: lysosomal protease promotes α-syn
Page 31 and 32: ROS, which may oxidize DA (a neurot
Page 33 and 34: together, although anti-oxidant act
Page 35 and 36: multicellular eukaryotic model orga
Page 37 and 38: C. elegans PD models C. elegans off
Page 39 and 40: Current studies To investigate gene
Page 41 and 42: Specific outcomes and future direct
Page 43 and 44: Corti, O., Hampe, C., Koutnikova, H
Page 45 and 46: Lautier, C., Goldwurm, S., Dürr, A
Page 47 and 48: Scherzer, C.R., Jensen, R.V., Gulla
Page 49 and 50: Willcox, B.J., Donlon, T.A., He, Q.
Page 51 and 52: Table 1.2. Summary of selected inve
Page 53 and 54: FIGURE LEGENDS Fig. 1.1. Schematic
Page 55 and 56: ABSTRACT Genomic multiplication of
Page 57 and 58:
et al., 2003). Maintenance of DA ne
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Pdat-1::FLAG-R05D11.6, Punc-54::DsR
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Biochemicals) and 1:10000 horseradi
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was washed with 100 µl RNase-free
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and effectively discerned via RNAi
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protein. The primary RNAi screen of
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expressing a polyglutamine::GFP fus
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stage (Fig. 2.2B-D). Two genes exhi
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with strong expression localized to
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Our data demonstrating that the C.
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disparate datasets, such as those o
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Driscoll, M., Gerstbrein, B. (2003)
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Singleton, A.B., Farrer, M., Johnso
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Table 2.2. Bioinformatic associatio
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Table 2.4. Results of all genes kno
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C09G12.9 Tumor susceptibility gene
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C32B5.13 Cathepsin H precursor C32B
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D1014.3 Alpha-soluble NSF attachmen
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F22B7.5 DNAJA3, mitochondrial precu
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F40F4.5 Tubulin alpha-ubiquitous ch
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F54F7.5 Serine/threonine kinase 24
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K07A9.2 CAMK1 K07C11.9 Hypothetical
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R74.4 DNAJB1 precursor T01B7.4 Pept
Page 103 and 104:
T22D1.9 PSMD2 T22F3.2 Ubiquitin spe
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Y38F2AL.3 Vacuolar ATP synthase sub
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Y73C8C.8 CENPE variant protein (Fra
Page 109 and 110:
Table 2.5. Summary of RNAi knockdow
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Figure 2.2 91
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Figure 2.4 93
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Figure 2.6 95
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FIGURE LEGENDS Figure 2.1. RNAi kno
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came from genes that were co-expres
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ABSTRACT Multiple Parkinson disease
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vacuoles (mammalian equivalent of l
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neuroprotection when the animals we
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To investigate the genetic interact
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illustrated that α-syn overexpress
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main contributor of α-syn phosphor
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Hoozemans, J.J., van Haastert, E.S.
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Figure 3.2 115
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FIGURE LEGENDS Figure 3.1. RAB3A, R
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CHAPTER FOUR RNA INTERFERENCE SCREE
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INTRODUCTION Central to Parkinson d
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thermotolerance (Lithgow et al., 19
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mCherry were transferred onto NGM p
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20°C for 72 hrs. The gravid adults
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pathways downstream of DAF-2 exclud
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were assayed by growing the RNAi-tr
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while DOG treatment enhanced α-syn
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enhance α-syn misfolding in a daf-
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REFERENCES Bar-On, P., Crews, L., K
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Lesage, S., Brice, A. (2009) Hum Mo
Page 161 and 162:
Table 4.1. Summary of genes analyze
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C12D12.2 Excitatory amino acid tran
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C47B2.2 n/a C47C12.6 Amiloride-sens
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F18A1.5 Replication protein A F18E3
Page 169 and 170:
F46E10.10 Malate dehydrogenase F46E
Page 171 and 172:
K08H10.1 Dentin sialophosphoprotein
Page 173 and 174:
T05A10.5 Cysteine-rich secretory pr
Page 175 and 176:
W03G9.6 Platelet-activating factor
Page 177 and 178:
Y58G8A.1 Acetylcholine receptor sub
Page 179 and 180:
Table 4.2. Summary of positive gene
Page 181 and 182:
Figure 4.1 161
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Figure 4.3 163
Page 185 and 186:
Figure 4.5 165
Page 187 and 188:
Figure 4.7 167
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Figure 4.4. Graph illustrating the
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in yeast but not in humans. In yeas
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kinase that has also been shown to
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mir-2/mir-43/mir-250/mir-797 superf
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of djr-1.1 and djr-1.2 enhance α-s
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expensive, since they require high
Page 201 and 202:
Ambros, V. (2001) Cell 107, 823-826
Page 203 and 204:
Yi, J., Tang, X.M. (1999) Cell Res
Page 205 and 206:
Figure 5.2 185
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