α-Synuclein (α-Syn) is a major component of protein inclusions known as Lewy bodies, which are hallmarks of synucleinopathies such as Parkinson's disease (PD). The α-Syn gene is one of the familial PD-causing genes and is also associated with an increased risk of sporadic PD. Numerous studies using α-Syn expressing transgenic animals have indicated that α-Syn plays a critical role in the common pathogenesis of synucleinopathies. Drosophila melanogaster has several advantages for modeling human neurodegenerative diseases and is widely used for studying their pathomechanisms and therapies. In fact, Drosophila models expressing α-Syn have already been established and proven to replicate several features of human PD. In this paper, we review the current research on synucleinopathies using α-Syn Drosophila models and, moreover, explore the possibilities of these models for comprehensive genetic analyses and large-scale drug screening towards elucidating the molecular pathogenesis and developing therapies for synucleinopathies. 1. Introduction Protein inclusions known as Lewy Bodies (LBs) are one of the hallmarks of Parkinson’s disease (PD), in which the major component is now known to be α-synuclein (α-Syn) [1, 2]. LBs are found in the substantia nigra in PD and also more extensively in other brain regions in other synucleinopathies including multiple system atrophy and dementia with Lewy bodies (DLB) [3, 4]. The α-Syn encoding gene, SNCA, is the first gene in which missense mutations such as A30P and A53T were found to cause familial PD [5, 6]. Furthermore, the multiplication mutations of α-Syn gene were also found to cause familial PD [7]. Most importantly, single nucleotide polymorphisms (SNPs) of α-Syn have been reported to associate with an increased risk of sporadic PD, which comprises the majority of PD patients [8–11]. α-Syn expression has been experimentally shown to mimic several aspects of PD in transgenic animals, such as motor dysfunction, α-Syn aggregation/accumulation, and neurodegeneration [12–14]. These phenotypes are manifested not only by mutations in the α-Syn gene but also by overexpression of wild-type α-Syn [15], indicating that α-Syn plays a critical role in the common pathogenesis of synucleinopathies. Drosophila melanogaster, commonly known as the fruit fly, has been recognized as a powerful organism for modeling human neurodegenerative diseases [16]. At least ~75% of human disease genes have Drosophila homologues [17]. Using Drosophila for modeling human neurodegenerative diseases has various advantages as follows: (1) analysis of
References
[1]
M. G. Spillantini, R. A. Crowther, R. Jakes, M. Hasegawa, and M. Goedert, “ -synuclein in filamentous inclusions of Lewy bodies from Parkinson's disease and dementia with Lewy bodies,” Proceedings of the National Academy of Sciences of the United States of America, vol. 95, no. 11, pp. 6469–6473, 1998.
[2]
M. G. Spillantini, M. L. Schmidt, V. M. Y. Lee, J. Q. Trojanowski, R. Jakes, and M. Goedert, “ -synuclein in Lewy bodies,” Nature, vol. 388, no. 6645, pp. 839–840, 1997.
[3]
K. Beyer and A. Ariza, “Protein aggregation mechanisms in synucleinopathies: commonalities and differences,” Journal of Neuropathology and Experimental Neurology, vol. 66, no. 11, pp. 965–974, 2007.
[4]
T. Iwatsubo, “Pathological biochemistry of -synucleinopathy,” Neuropathology, vol. 27, no. 5, pp. 474–478, 2007.
[5]
R. Krüger, W. Kuhn, T. Müller et al., “Ala30Pro mutation in the gene encoding -synuclein in Parkinson's disease,” Nature Genetics, vol. 18, no. 2, pp. 106–108, 1998.
[6]
M. H. Polymeropoulos, C. Lavedan, E. Leroy et al., “Mutation in the -synuclein gene identified in families with Parkinson's disease,” Science, vol. 276, no. 5321, pp. 2045–2047, 1997.
[7]
A. B. Singleton, M. Farrer, J. Johnson et al., “ -synuclein locus triplication causes Parkinson's disease,” Science, vol. 302, no. 5646, p. 841, 2003.
[8]
J. C. Mueller, J. Fuchs, A. Hofer et al., “Multiple regions of -synuclein are associated with Parkinson's disease,” Annals of Neurology, vol. 57, no. 4, pp. 535–541, 2005.
[9]
I. Mizuta, W. Satake, Y. Nakabayashi et al., “Multiple candidate gene analysis identifies -synuclein as a susceptibility gene for sporadic Parkinson's disease,” Human Molecular Genetics, vol. 15, no. 7, pp. 1151–1158, 2006.
[10]
W. Satake, Y. Nakabayashi, I. Mizuta et al., “Genome-wide association study identifies common variants at four loci as genetic risk factors for Parkinson's disease,” Nature Genetics, vol. 41, no. 12, pp. 1303–1307, 2009.
[11]
J. Simón-Sánchez, C. Schulte, J. M. Bras et al., “Genome-wide association study reveals genetic risk underlying Parkinson's disease,” Nature Genetics, vol. 41, no. 12, pp. 1308–1312, 2009.
[12]
M. B. Feany and W. W. Bender, “A Drosophila model of Parkinson's disease,” Nature, vol. 404, no. 6776, pp. 394–398, 2000.
[13]
M. K. Lee, W. Stirling, Y. Xu et al., “Human -synuclein-harboring familial Parkinson's disease-linked Ala-53 → Thr mutation causes neurodegenerative disease with -synuclein aggregation in transgenic mice,” Proceedings of the National Academy of Sciences of the United States of America, vol. 99, no. 13, pp. 8968–8973, 2002.
[14]
E. Masliah, E. Rockenstein, I. Veinbergs et al., “Dopaminergic loss and inclusion body formation in -synuclein mice: implications for neurodegenerative disorders,” Science, vol. 287, no. 5456, pp. 1265–1269, 2000.
[15]
M. Farrer, J. Kachergus, L. Forno et al., “Comparison of kindreds with Parkinsonism and -synuclein genomic multiplications,” Annals of Neurology, vol. 55, no. 2, pp. 174–179, 2004.
[16]
J. Bilen and N. M. Bonini, “Drosophila as a model for human neurodegenerative disease,” Annual Review of Genetics, vol. 39, pp. 153–171, 2005.
[17]
L. T. Reiter, L. Potocki, S. Chien, M. Gribskov, and E. Bier, “A systematic analysis of human disease-associated gene sequences in Drosophila melanogaster,” Genome Research, vol. 11, no. 6, pp. 1114–1125, 2001.
[18]
D. P. Karpinar, M. B. G. Balija, S. Kügler et al., “Pre-fibrillar -synuclein variants with impaired -structure increase neurotoxicity in parkinson's disease models,” EMBO Journal, vol. 28, no. 20, pp. 3256–3268, 2009.
[19]
G. Du, X. Liu, X. Chen et al., “Drosophila histone deacetylase 6 protects dopaminergic neurons against -synuclein toxicity by promoting inclusion formation,” Molecular Biology of the Cell, vol. 21, no. 13, pp. 2128–2137, 2010.
[20]
P. K. Auluck, H. Y. E. Chan, J. Q. Trojanowski, V. M. Y. Lee, and N. M. Bonini, “Chaperone suppression of -synuclein toxicity in a Drosophila model for Parkinson's disease,” Science, vol. 295, no. 5556, pp. 865–868, 2002.
[21]
P. K. Auluck and N. M. Bonini, “Pharmacological prevention of Parkinson disease in Drosophila,” Nature Medicine, vol. 8, no. 11, pp. 1185–1186, 2002.
[22]
F. K. M. Lee, A. K. Y. Wong, Y. W. Lee, OI. W. Wan, H. Y. Edwin Chan, and K. K. K. Chung, “The role of ubiquitin linkages on -synuclein induced-toxicity in a Drosophila model of Parkinson's disease,” Journal of Neurochemistry, vol. 110, no. 1, pp. 208–219, 2009.
[23]
V. Cullen, M. Lindfors, J. Ng et al., “Cathepsin D expression level affects alpha-synuclein processing, aggregation, and toxicity in vivo,” Molecular Brain, vol. 2, no. 1, article 5, 2009.
[24]
LI. Chen and M. B. Feany, “ -synuclein phosphorylation controls neurotoxicity and inclusion formation in a Drosophila model of Parkinson disease,” Nature Neuroscience, vol. 8, no. 5, pp. 657–663, 2005.
[25]
L. Chen, M. Periquet, X. Wang et al., “Tyrosine and serine phosphorylation of -synuclein have opposing effects on neurotoxicity and soluble oligomer formation,” Journal of Clinical Investigation, vol. 119, no. 11, pp. 3257–3265, 2009.
[26]
M. Periquet, T. Fulga, L. Myllykangas, M. G. Schlossmacher, and M. B. Feany, “Aggregated -synuclein mediates dopaminergic neurotoxicity in vivo,” Journal of Neuroscience, vol. 27, no. 12, pp. 3338–3346, 2007.
[27]
B. M. Dufty, L. R. Warner, S. T. Hou et al., “Calpain-cleavage of -synuclein: connecting proteolytic processing to disease-linked aggregation,” American Journal of Pathology, vol. 170, no. 5, pp. 1725–1738, 2007.
[28]
J. A. Botella, F. Bayersdorfer, and S. Schneuwly, “Superoxide dismutase overexpression protects dopaminergic neurons in a Drosophila model of Parkinson's disease,” Neurobiology of Disease, vol. 30, no. 1, pp. 65–73, 2008.
[29]
F. Bayersdorfer, A. Voigt, S. Schneuwly, and J. A. Botella, “Dopamine-dependent neurodegeneration in Drosophila models of familial and sporadic Parkinson's disease,” Neurobiology of Disease, vol. 40, no. 1, pp. 113–119, 2010.
[30]
K. Trinh, K. Moore, P. D. Wes et al., “Induction of the phase II detoxification pathway suppresses neuron loss in Drosophila models of Parkinson's disease,” Journal of Neuroscience, vol. 28, no. 2, pp. 465–472, 2008.
[31]
H. Jia, X. Li, H. Gao et al., “High doses of nicotinamide prevent oxidative mitochondrial dysfunction in a cellular model and improve motor deficit in a Drosophila model of Parkinson's disease,” Journal of Neuroscience Research, vol. 86, no. 9, pp. 2083–2090, 2008.
[32]
J. Long, H. Gao, L. Sun, J. Liu, and X. Zhao-Wilson, “Grape extract protects mitochondria from oxidative damage and improves locomotor dysfunction and extends lifespan in a Drosophila Parkinson's disease model,” Rejuvenation Research, vol. 12, no. 5, pp. 321–331, 2009.
[33]
A. F. M. Haywood and B. E. Staveley, “Parkin counteracts symptoms in a Drosophila model of Parkinson's disease,” BMC Neuroscience, vol. 5, article 14, 2004.
[34]
A. F. M. Haywood and B. E. Staveley, “Mutant -synuclein-induced degeneration is reduced by parkin in a fly model of Parkinson's disease,” Genome, vol. 49, no. 5, pp. 505–510, 2006.
[35]
Y. Yang, I. Nishimura, Y. Imai, R. Takahashi, and B. Lu, “Parkin suppresses dopaminergic neuron-selective neurotoxicity induced by Pael-R in Drosophila,” Neuron, vol. 37, no. 6, pp. 911–924, 2003.
[36]
A. M. Todd and B. E. Staveley, “Pink1 suppresses -synuclein-induced phenotypes in a Drosophila model of Parkinson's disease,” Genome, vol. 51, no. 12, pp. 1040–1046, 2008.
[37]
V. N. Uversky, “Neuropathology, biochemistry, and biophysics of -synuclein aggregation,” Journal of Neurochemistry, vol. 103, no. 1, pp. 17–37, 2007.
[38]
H. Naiki and Y. Nagai, “Molecular pathogenesis of protein misfolding diseases: pathological molecular environments versus quality control systems against misfolded proteins,” Journal of Biochemistry, vol. 146, no. 6, pp. 751–756, 2009.
[39]
J. C. Young, V. R. Agashe, K. Siegers, and F. U. Hartl, “Pathways of chaperone-mediated protein folding in the cytosol,” Nature Reviews Molecular Cell Biology, vol. 5, no. 10, pp. 781–791, 2004.
[40]
D. C. Rubinsztein, “The roles of intracellular protein-degradation pathways in neurodegeneration,” Nature, vol. 443, no. 7113, pp. 780–786, 2006.
[41]
R. Bandopadhyay and J. de Belleroche, “Pathogenesis of Parkinson's disease: emerging role of molecular chaperones,” Trends in Molecular Medicine, vol. 16, no. 1, pp. 27–36, 2010.
[42]
P. J. McLean, H. Kawamata, S. Shariff et al., “TorsinA and heat shock proteins act as molecular chaperones: suppression of -synuclein aggregation,” Journal of Neurochemistry, vol. 83, no. 4, pp. 846–854, 2002.
[43]
T. F. Outeiro, J. Klucken, K. E. Strathearn et al., “Small heat shock proteins protect against -synuclein-induced toxicity and aggregation,” Biochemical and Biophysical Research Communications, vol. 351, no. 3, pp. 631–638, 2006.
[44]
M. M. Dedmon, J. Christodoulou, M. R. Wilson, and C. M. Dobson, “Heat shock protein 70 inhibits -synuclein fibril formation via preferential binding to prefibrillar species,” Journal of Biological Chemistry, vol. 280, no. 15, pp. 14733–14740, 2005.
[45]
J. Klucken, Y. Shin, E. Masliah, B. T. Hyman, and P. J. McLean, “Hsp70 reduces -synuclein aggregation and toxicity,” Journal of Biological Chemistry, vol. 279, no. 24, pp. 25497–25502, 2004.
[46]
A. Zourlidou, M. D. Payne Smith, and D. S. Latchman, “HSP27 but not HSP70 has a potent protective effect against -synuclein-induced cell death in mammalian neuronal cells,” Journal of Neurochemistry, vol. 88, no. 6, pp. 1439–1448, 2004.
[47]
T. Pan, S. Kondo, W. Le, and J. Jankovic, “The role of autophagy-lysosome pathway in neurodegeneration associated with Parkinson's disease,” Brain, vol. 131, no. 8, pp. 1969–1978, 2008.
[48]
J. Tyynel？, I. Sohar, D. E. Sleat et al., “A mutation in the ovine cathepsin D gene causes a congenital lysosomal storage disease with profound neurodegeneration,” EMBO Journal, vol. 19, no. 12, pp. 2786–2792, 2000.
[49]
D. Sevlever, P. Jiang, and S. H. C. Yen, “Cathepsin D is the main lysosomal enzyme involved in the degradation of -synuclein and generation of its carboxy-terminally truncated species,” Biochemistry, vol. 47, no. 36, pp. 9678–9687, 2008.
[50]
H. Fujiwara, M. Hasegawa, N. Dohmae et al., “ -synuclein is phosphorylated in synucleinopathy lesions,” Nature Cell Biology, vol. 4, no. 2, pp. 160–164, 2002.
[51]
M. Takahashi, H. Kanuka, H. Fujiwara et al., “Phosphorylation of -synuclein characteristic of synucleinopathy lesions is recapitulated in -synuclein transgenic Drosophila,” Neuroscience Letters, vol. 336, no. 3, pp. 155–158, 2003.
[52]
M. Baba, S. Nakajo, P. H. Tu et al., “Aggregation of -synuclein in Lewy bodies of sporadic Parkinson's disease and dementia with Lewy bodies,” American Journal of Pathology, vol. 152, no. 4, pp. 879–884, 1998.
[53]
W. P. Gai, J. H. T. Power, P. C. Blumbergs, J. G. Culvenor, and P. H. Jensen, “ -synuclein immunoisolation of glial inclusions from multiple system atrophy brain tissue reveals multiprotein components,” Journal of Neurochemistry, vol. 73, no. 5, pp. 2093–2100, 1999.
[54]
I. V. J. Murray, B. I. Giasson, S. M. Quinn et al., “Role of -synuclein carboxy-terminus on fibril formation in vitro,” Biochemistry, vol. 42, no. 28, pp. 8530–8540, 2003.
[55]
L. C. Serpell, J. Berriman, R. Jakes, M. Goedert, and R. A. Crowther, “Fiber diffraction of synthetic -synuclein filaments shows amyloid-like cross-β conformation,” Proceedings of the National Academy of Sciences of the United States of America, vol. 97, no. 9, pp. 4897–4902, 2000.
[56]
A. M. Bodles, D. J. S. Guthrie, B. Greer, and G. Brent Irvine, “Identification of the region of non-Aβ component (NAC) of Alzheimer's disease amyloid responsible for its aggregation and toxicity,” Journal of Neurochemistry, vol. 78, no. 2, pp. 384–395, 2001.
[57]
B. I. Giasson, I. V. J. Murray, J. Q. Trojanowski, and V. M. Y. Lee, “A hydrophobic stretch of 12 amino acid residues in the middle of -synuclein is essential for filament assembly,” Journal of Biological Chemistry, vol. 276, no. 4, pp. 2380–2386, 2001.
[58]
A. J. Mishizen-Eberz, R. P. Guttmann, B. I. Giasson et al., “Distinct cleavage patterns of normal and pathologic forms of -synuclein by calpain I in vitro,” Journal of Neurochemistry, vol. 86, no. 4, pp. 836–847, 2003.
[59]
A. J. Mishizen-Eberz, E. H. Norris, B. I. Giasson et al., “Cleavage of -synuclein by calpain: potential role in degradation of fibrillized and nitrated species of -synuclein,” Biochemistry, vol. 44, no. 21, pp. 7818–7829, 2005.
[60]
W. Li, N. West, E. Colla et al., “Aggregation promoting C-terminal truncation of -synuclein is a normal cellular process and is enhanced by the familial Parkinson's disease-linked mutations,” Proceedings of the National Academy of Sciences of the United States of America, vol. 102, no. 6, pp. 2162–2167, 2005.
[61]
T. Kitada, S. Asakawa, N. Hattori et al., “Mutations in the parkin gene cause autosomal recessive juvenile parkinsonism,” Nature, vol. 392, no. 6676, pp. 605–608, 1998.
[62]
E. M. Valente, P. M. Abou-Sleiman, V. Caputo et al., “Hereditary early-onset Parkinson's disease caused by mutations in PINK1,” Science, vol. 304, no. 5674, pp. 1158–1160, 2004.
[63]
I. E. Clark, M. W. Dodson, C. Jiang et al., “Drosophila pink1 is required for mitochondrial function and interacts genetically with parkin,” Nature, vol. 441, no. 7097, pp. 1162–1166, 2006.
[64]
J. Park, S. B. Lee, S. Lee et al., “Mitochondrial dysfunction in Drosophila PINK1 mutants is complemented by parkin,” Nature, vol. 441, no. 7097, pp. 1157–1161, 2006.
[65]
C. R. Scherzer, R. V. Jensen, S. R. Gullans, and M. B. Feany, “Gene expression changes presage neurodegeneration in a Drosophila model of Parkinson's disease,” Human Molecular Genetics, vol. 12, no. 19, pp. 2457–2466, 2003.
[66]
Z. Xun, R. A. Sowell, T. C. Kaufman, and D. E. Clemmer, “Lifetime proteomic profiling of an A30P -synuclein Drosophila model of Parkinson's disease,” Journal of Proteome Research, vol. 6, no. 9, pp. 3729–3738, 2007.
[67]
Z. Xun, R. A. Sowell, T. C. Kaufman, and D. E. Clemmer, “Quantitative proteomics of a presymptomatic A53T -synuclein Drosophila model of Parkinson disease,” Molecular and Cellular Proteomics, vol. 7, no. 7, pp. 1191–1203, 2008.
[68]
P. Fernandez-Funez, M. L. Nino-Rosales, B. de Gouyon et al., “Identification of genes that modify ataxin-1-induced neurodegeneration,” Nature, vol. 408, no. 6808, pp. 101–106, 2000.
[69]
R. G. Pendleton, F. Parvez, M. Sayed, and R. Hillman, “Effects of pharmacological agents upon a transgenic model of Parkinson's disease in Drosophila melanogaster,” Journal of Pharmacology and Experimental Therapeutics, vol. 300, no. 1, pp. 91–96, 2002.
[70]
E. Kontopoulos, J. D. Parvin, and M. B. Feany, “ -synuclein acts in the nucleus to inhibit histone acetylation and promote neurotoxicity,” Human Molecular Genetics, vol. 15, no. 20, pp. 3012–3023, 2006.
[71]
T. F. Outeiro, E. Kontopoulos, S. M. Altmann et al., “Sirtuin 2 inhibitors rescue -synuclein-mediated toxicity in models of Parkinson's disease,” Science, vol. 317, no. 5837, pp. 516–519, 2007.
