Abstract
Parkinson’s disease is the second most common neurodegenerative disease without cure. It is characterized by α-synuclein accumulation and aggregation in dopaminergic and other types of neurons. Because α-synuclein accumulation leads to a toxic gain of function, its ectopic expression in Drosophila has been a useful in vivo model for testing modifiers of its toxicity. This chapter describes four assays: the rapid iterative negative geotaxis, rough eye phenotype, quantification of dopaminergic neuronal loss, and measurements of circadian effects.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
References
Tofaris GK, Spillantini MG (2005) Alpha-synuclein dysfunction in Lewy body diseases. Mov Disord 20(Suppl 12):S37–S44
Tofaris GK (2012) Lysosome-dependent pathways as a unifying theme in Parkinson’s disease. Mov Disord 27(11):1364–1369
Reiter LT, Potocki L, Chien S, Gribskov M, Bier E (2001) A systematic analysis of human disease-associated gene sequences in Drosophila melanogaster. Genome Res 11(6):1114–1125
Brand AH, Perrimon N (1993) Targeted gene expression as a means of altering cell fates and generating dominant phenotypes. Development 118(2):401–415
Dietzl G, Chen D, Schnorrer F, Su KC, Barinova Y, Fellner M, Gasser B, Kinsey K, Oppel S, Scheiblauer S, Couto A, Marra V, Keleman K, Dickson BJ (2007) A genome-wide transgenic RNAi library for conditional gene inactivation in Drosophila. Nature 448(7150):151–156
Greene JC, Whitworth AJ, Kuo I, Andrews LA, Feany MB, Pallanck LJ (2003) Mitochondrial pathology and apoptotic muscle degeneration in Drosophila parkin mutants. Proc Natl Acad Sci U S A 100(7):4078–4083
Yang Y, Gehrke S, Imai Y, Huang Z, Ouyang Y, Wang JW, Yang L, Beal MF, Vogel H, Lu B (2006) Mitochondrial pathology and muscle and dopaminergic neuron degeneration caused by inactivation of Drosophila Pink1 is rescued by Parkin. Proc Natl Acad Sci U S A 103(28):10793–10798
Clark IE, Dodson MW, Jiang C, Cao JH, Huh JR, Seol JH, Yoo SJ, Hay BA, Guo M (2006) Drosophila pink1 is required for mitochondrial function and interacts genetically with parkin. Nature 441(7097):1162–1166
Park J, Lee SB, Lee S, Kim Y, Song S, Kim S, Bae E, Kim J, Shong M, Kim JM, Chung J (2006) Mitochondrial dysfunction in Drosophila PINK1 mutants is complemented by parkin. Nature 441(7097):1157–1161
McWilliams TG, Muqit MM (2017) PINK1 and Parkin: emerging themes in mitochondrial homeostasis. Curr Opin Cell Biol 45:83–91
Dodson MW, Zhang T, Jiang C, Chen S, Guo M (2012) Roles of the Drosophila LRRK2 homolog in Rab7-dependent Hum Mol lysosomal positioning. Hum Mol Genet 21(6):1350–1363
MacLeod DA, Rhinn H, Kuwahara T, Zolin A, Di Paolo G, McCabe BD, Marder KS, Honig LS, Clark LN, Small SA, Abeliovich A (2013) RAB7L1 interacts with LRRK2 to modify intraneuronal protein sorting and Parkinson’s disease risk. Neuron 77(3):425–439
Soukup SF, Kuenen S, Vanhauwaert R, Manetsberger J, Hernández-Díaz S, Swerts J, Schoovaerts N, Vilain S, Gounko NV, Vints K, Geens A, De Strooper B, Verstreken P (2016) A LRRK2-dependent endophilin A phosphoswitch is critical for macroautophagy at presynaptic terminals. Neuron 92(4):829–844
Tofaris GK, Goedert M, Spillantini MG (2017) The transcellular propagation and intracellular trafficking of α-synuclein. Cold Spring Harb Perspect Med 7(9). pii: a024380). https://doi.org/10.1101/cshperspect.a024380
Feany MB, Bender WW (2000) A Drosophila model of Parkinson’s disease. Nature 404(6776):394–398
Davies SE, Hallett PJ, Moens T, Smith G, Mangano E, Kim HT, Goldberg AL, Liu JL, Isacson O, Tofaris GK (2014) Enhanced ubiquitin-dependent degradation by Nedd4 protects against α-synuclein accumulation and toxicity in animal models of Parkinson’s disease. Neurobiol Dis 64:79–87
Alexopoulou Z, Lang J, Perrett RM, Elschami M, Hurry ME, Kim HT, Mazaraki D, Szabo A, Kessler BM, Goldberg AL, Ansorge O, Fulga TA, Tofaris GK (2016) Deubiquitinase Usp8 regulates α-synuclein clearance and modifies its toxicity in Lewy body disease. Proc Natl Acad Sci U S A 113(32):E4688–E4697
Szabó A, Papin C, Zorn D, Ponien P, Weber F, Raabe T, Rouyer F (2013) The CK2 kinase stabilizes CLOCK and represses its activity in the Drosophila circadian oscillator. PLoS Biol 11(8):e1001645
Breen DP, Vuono R, Nawarathna U, Fisher K, Shneerson JM, Reddy AB, Barker RA (2014) Sleep and circadian rhythm regulation in early Parkinson disease. JAMA Neurol 71(5):589–595
Gajula Balija MB, Griesinger C, Herzig A, Zweckstetter M, Jäckle H (2011) Pre-fibrillar α-synuclein mutants cause Parkinson’s disease-like non-motor symptoms in Drosophila. PLoS One 6(9):e24701
Karpinar DP, Balija MB, Kügler S, Opazo F, Rezaei-Ghaleh N, Wender N, Kim HY, Taschenberger G, Falkenburger BH, Heise H, Kumar A, Riedel D, Fichtner L, Voigt A, Braus GH, Giller K, Becker S, Herzig A, Baldus M, Jäckle H, Eimer S, Schulz JB, Griesinger C, Zweckstetter M (2009) Pre-fibrillar alpha-synuclein variants with impaired beta-structure increase neurotoxicity in Parkinson’s disease models. EMBO J 28(20):3256–3268
Gargano JW, Martin I, Bhandari P, Grotewiel MS (2005) Rapid iterative negative geotaxis (RING): a new method for assessing age-related locomotor decline in Drosophila. Exp Gerontol 40(5):386–395
Acknowledgements
GKT is funded by a Wellcome Trust Intermediate Clinical Fellowship (097479/Z/11/Z), the Wellcome Beit Prize (097479/Z/11/A), and the NIHR Oxford Biomedical Research Centre.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2019 Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Szabo, A., Tofaris, G.K. (2019). Monitoring α-Synuclein Proteotoxicity in Drosophila Models. In: Bartels, T. (eds) Alpha-Synuclein. Methods in Molecular Biology, vol 1948. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-9124-2_15
Download citation
DOI: https://doi.org/10.1007/978-1-4939-9124-2_15
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-9123-5
Online ISBN: 978-1-4939-9124-2
eBook Packages: Springer Protocols