Nigral overexpression of alpha-synuclein in the absence of parkin enhances alpha-synuclein phosphorylation but does not modulate dopaminergic neurodegeneration

Molecular Neurodegeneration

Veerle Baekelandt²  Chris Van den Haute³  Olga Corti¹  Anke Van der Perren²  Marusela Oliveras-Salvá²  Anne-Sophie Van Rompuy² 

[1] CNRS, UMR 7225, Paris, F-75013, France;Laboratory for Neurobiology and Gene Therapy, Department of Neurosciences, KU Leuven, Flanders, Belgium;Leuven Viral Vector Core, KU Leuven, Leuven, 3000, Belgium

关键词: Knockout;    Adeno-associated viral vectors;    Parkin;    Alpha-synuclein phosphorylation;    Alpha-synuclein;    Parkinson’s disease;   

Others  :  1216346
DOI  :  10.1186/s13024-015-0017-8

 received in 2013-09-19, accepted in 2015-04-10,  发布年份 2015

【 摘 要 】

Background

Alpha-synuclein is a key protein in the pathogenesis of Parkinson’s disease. Mutations in the parkin gene are the most common cause of early-onset autosomal recessive Parkinson’s disease, probably through a loss-of-function mechanism. However, the molecular mechanism by which loss of parkin function leads to the development of the disease and the role of alpha-synuclein in parkin-associated Parkinson’s disease is still not elucidated. Conflicting results were reported about the effect of the absence of parkin on alpha-synuclein-mediated neurotoxicity using a transgenic approach. In this study, we investigated the effect of loss of parkin on alpha-synuclein neuropathology and toxicity in adult rodent brain using viral vectors. Therefore, we overexpressed human wild type alpha-synuclein in the substantia nigra of parkin knockout and wild type mice using two different doses of recombinant adeno-associated viral vectors.

Results

No difference was observed in nigral dopaminergic cell loss between the parkin knockout mice and wild type mice up to 16 weeks after viral vector injection. However, the level of alpha-synuclein phosphorylated at serine residue 129 in the substantia nigra was significantly increased in the parkin knockout mice compared to the wild type mice while the total expression level of alpha-synuclein was similar in both groups. The increased alpha-synuclein phosphorylation was confirmed in a parkin knockdown cell line.

Conclusions

These findings support a functional relationship between parkin and alpha-synuclein phosphorylation in rodent brain.

【 授权许可】

   
2015 Rompuy et al.

【 图 表 】

Figure 1.

Figure 2.

Figure 3.

Figure 4.

Figure 5.

Figure 6.

Figure 7.

【 参考文献 】

• [1]Lees AJ, Hardy J, Revesz T. Parkinson's disease. Lancet. 2009; 373:2055-2066.
• [2]Schulte C, Gasser T. Genetic basis of Parkinson's disease: inheritance, penetrance, and expression. Appl Clin Genet. 2011; 4:67-80.
• [3]Maroteaux L, Campanelli JT, Scheller RH. Synuclein: a neuron-specific protein localized to the nucleus and presynaptic nerve terminal. J Neurosci. 1988; 8:2804-2815.
• [4]Abeliovich A, Schmitz Y, Farinas I, Choi-Lundberg D, Ho WH, Castillo PE et al.. Mice lacking alpha-synuclein display functional deficits in the nigrostriatal dopamine system. Neuron. 2000; 25:239-252.
• [5]Perez RG, Waymire JC, Lin E, Liu JJ, Guo F, Zigmond MJ. A role for alpha-synuclein in the regulation of dopamine biosynthesis. J Neurosci. 2002; 22:3090-3099.
• [6]George JM, Jin H, Woods WS, Clayton DF. Characterization of a novel protein regulated during the critical period for song learning in the zebra finch. Neuron. 1995; 15:361-372.
• [7]Lotharius J, Brundin P. Pathogenesis of Parkinson's disease: dopamine, vesicles and alpha-synuclein. Nat Rev Neurosci. 2002; 3:932-942.
• [8]Deleersnijder A, Gerard M, Debyser Z, Baekelandt V. The remarkable conformational plasticity of alpha-synuclein: blessing or curse? Trends Mol Med. 2013; 19:368-377.
• [9]Spillantini MG, Schmidt ML, Lee VM, Trojanowski JQ, Jakes R, Goedert M. Alpha-synuclein in Lewy bodies. Nature. 1997; 388:839-840.
• [10]Chesselet MF. In vivo alpha-synuclein overexpression in rodents: a useful model of Parkinson's disease? Exp Neurol. 2008; 209:22-27.
• [11]Kitada T, Asakawa S, Hattori N, Matsumine H, Yamamura Y, Minoshima S et al.. Mutations in the parkin gene cause autosomal recessive juvenile parkinsonism. Nature. 1998; 392:605-608.
• [12]Imai Y, Soda M, Takahashi R. Parkin suppresses unfolded protein stress-induced cell death through its E3 ubiquitin-protein ligase activity. J Biol Chem. 2000; 275:35661-35664.
• [13]Shimura H, Hattori N, Kubo S, Mizuno Y, Asakawa S, Minoshima S et al.. Familial Parkinson disease gene product, parkin, is a ubiquitin-protein ligase. Nat Genet. 2000; 25:302-305.
• [14]Zhang Y, Gao J, Chung KK, Huang H, Dawson VL, Dawson TM. Parkin functions as an E2-dependent ubiquitin- protein ligase and promotes the degradation of the synaptic vesicle-associated protein, CDCrel-1. Proc Natl Acad Sci U S A. 2000; 97:13354-13359.
• [15]Exner N, Lutz AK, Haass C, Winklhofer KF. Mitochondrial dysfunction in Parkinson's disease: molecular mechanisms and pathophysiological consequences. EMBO J. 2012; 31:3038-3062.
• [16]Fallon L, Belanger CM, Corera AT, Kontogiannea M, Regan-Klapisz E, Moreau F et al.. A regulated interaction with the UIM protein Eps15 implicates parkin in EGF receptor trafficking and PI(3)K-Akt signalling. Nat Cell Biol. 2006; 8:834-842.
• [17]Henn IH, Bouman L, Schlehe JS, Schlierf A, Schramm JE, Wegener E et al.. Parkin mediates neuroprotection through activation of IkappaB kinase/nuclear factor-kappaB signaling. J Neurosci. 2007; 27:1868-1878.
• [18]Chung KK, Zhang Y, Lim KL, Tanaka Y, Huang H, Gao J et al.. Parkin ubiquitinates the alpha-synuclein-interacting protein, synphilin-1: implications for Lewy-body formation in Parkinson disease. Nat Med. 2001; 7:1144-1150.
• [19]Goldberg MS, Fleming SM, Palacino JJ, Cepeda C, Lam HA, Bhatnagar A et al.. Parkin-deficient mice exhibit nigrostriatal deficits but not loss of dopaminergic neurons. J Biol Chem. 2003; 278:43628-43635.
• [20]Ko HS, von Coelln R, Sriram SR, Kim SW, Chung KK, Pletnikova O et al.. Accumulation of the authentic parkin substrate aminoacyl-tRNA synthetase cofactor, p38/JTV-1, leads to catecholaminergic cell death. J Neurosci. 2005; 25:7968-7978.
• [21]Oliveras-Salva M, Van Rompuy AS, Heeman B, Van den Haute C, Baekelandt V. Loss-of-function rodent models for parkin and PINK1. J Parkinsons Dis. 2011; 1:229-251.
• [22]Doherty KM, Silveira-Moriyama L, Parkkinen L, Healy DG, Farrell M, Mencacci NE et al.. Parkin disease: a clinicopathologic entity? JAMA Neurol. 2013; 70:571-579.
• [23]Haywood AF, Staveley BE. Mutant alpha-synuclein-induced degeneration is reduced by parkin in a fly model of Parkinson's disease. Genome. 2006; 49:505-510.
• [24]Khandelwal PJ, Dumanis SB, Feng LR, Maguire-Zeiss K, Rebeck G, Lashuel HA et al.. Parkinson-related parkin reduces alpha-Synuclein phosphorylation in a gene transfer model. Mol Neurodegener. 2010; 5:47. BioMed Central Full Text
• [25]Lo Bianco C, Schneider BL, Bauer M, Sajadi A, Brice A, Iwatsubo T et al.. Lentiviral vector delivery of parkin prevents dopaminergic degeneration in an alpha-synuclein rat model of Parkinson's disease. Proc Natl Acad Sci U S A. 2004; 101:17510-17515.
• [26]Petrucelli L, O'Farrell C, Lockhart PJ, Baptista M, Kehoe K, Vink L et al.. Parkin protects against the toxicity associated with mutant alpha-synuclein: proteasome dysfunction selectively affects catecholaminergic neurons. Neuron. 2002; 36:1007-1019.
• [27]Yamada M, Mizuno Y, Mochizuki H. Parkin gene therapy for alpha-synucleinopathy: a rat model of Parkinson's disease. Hum Gene Ther. 2005; 16:262-270.
• [28]Yang Y, Nishimura I, Imai Y, Takahashi R, Lu B. Parkin suppresses dopaminergic neuron-selective neurotoxicity induced by Pael-R in Drosophila. Neuron. 2003; 37:911-924.
• [29]Yasuda T, Miyachi S, Kitagawa R, Wada K, Nihira T, Ren YR et al.. Neuronal specificity of alpha-synuclein toxicity and effect of Parkin co-expression in primates. Neuroscience. 2007; 144:743-753.
• [30]Fournier M, Vitte J, Garrigue J, Langui D, Dullin JP, Saurini F et al.. Parkin deficiency delays motor decline and disease manifestation in a mouse model of synucleinopathy. PLoS One. 2009; 4: Article ID e6629
• [31]von Coelln R, Thomas B, Andrabi SA, Lim KL, Savitt JM, Saffary R et al.. Inclusion body formation and neurodegeneration are parkin independent in a mouse model of alpha-synucleinopathy. J Neurosci. 2006; 26:3685-3696.
• [32]Stichel CC, Zhu XR, Bader V, Linnartz B, Schmidt S, Lubbert H. Mono- and double-mutant mouse models of Parkinson's disease display severe mitochondrial damage. Hum Mol Genet. 2007; 16:2377-2393.
• [33]Oliveras-Salva M, Van der Perren A, Casadei N, Stroobants S, Nuber S, D'Hooge R et al.. rAAV2/7 vector-mediated overexpression of alpha-synuclein in mouse substantia nigra induces protein aggregation and progressive dose-dependent neurodegeneration. Mol Neurodegener. 2013; 8:44. BioMed Central Full Text
• [34]Anderson JP, Walker DE, Goldstein JM, de Laat R, Banducci K, Caccavello RJ et al.. Phosphorylation of Ser-129 is the dominant pathological modification of alpha-synuclein in familial and sporadic Lewy body disease. J Biol Chem. 2006; 281:29739-29752.
• [35]Fujiwara H, Hasegawa M, Dohmae N, Kawashima A, Masliah E, Goldberg MS et al.. Alpha-Synuclein is phosphorylated in synucleinopathy lesions. Nat Cell Biol. 2002; 4:160-164.
• [36]Inglis KJ, Chereau D, Brigham EF, Chiou SS, Schobel S, Frigon NL et al.. Polo-like kinase 2 (PLK2) phosphorylates alpha-synuclein at serine 129 in central nervous system. J Biol Chem. 2009; 284:2598-2602.
• [37]Lee KW, Chen W, Junn E, Im JY, Grosso H, Sonsalla PK et al.. Enhanced phosphatase activity attenuates alpha-synucleinopathy in a mouse model. J Neurosci. 2011; 31:6963-6971.
• [38]Van der Perren A, Toelen J, Carlon M, Van den Haute C, Coun F, Heeman B et al.. Efficient and stable transduction of dopaminergic neurons in rat substantia nigra by rAAV 2/1, 2/2, 2/5, 2/6.2, 2/7, 2/8 and 2/9. Gene Ther. 2011; 18:517-527.
• [39]Lee MK, Stirling W, Xu Y, Xu X, Qui D, Mandir AS et al.. Human alpha-synuclein-harboring familial Parkinson's disease-linked Ala-53 – > Thr mutation causes neurodegenerative disease with alpha-synuclein aggregation in transgenic mice. Proc Natl Acad Sci U S A. 2002; 99:8968-8973.
• [40]Neumann M, Kahle PJ, Giasson BI, Ozmen L, Borroni E, Spooren W et al.. Misfolded proteinase K-resistant hyperphosphorylated alpha-synuclein in aged transgenic mice with locomotor deterioration and in human alpha-synucleinopathies. J Clin Invest. 2002; 110:1429-1439.
• [41]Itier JM, Ibanez P, Mena MA, Abbas N, Cohen-Salmon C, Bohme GA et al.. Parkin gene inactivation alters behaviour and dopamine neurotransmission in the mouse. Hum Mol Genet. 2003; 12:2277-2291.
• [42]Martella G, Platania P, Vita D, Sciamanna G, Cuomo D, Tassone A et al.. Enhanced sensitivity to group II mGlu receptor activation at corticostriatal synapses in mice lacking the familial parkinsonism-linked genes PINK1 or Parkin. Exp Neurol. 2009; 215:388-396.
• [43]Shin JH, Ko HS, Kang H, Lee Y, Lee YI, Pletinkova O et al.. PARIS (ZNF746) repression of PGC-1alpha contributes to neurodegeneration in Parkinson's disease. Cell. 2011; 144:689-702.
• [44]Li J, Uversky VN, Fink AL. Conformational behavior of human alpha-synuclein is modulated by familial Parkinson's disease point mutations A30P and A53T. Neurotoxicology. 2002; 23:553-567.
• [45]Jo E, Fuller N, Rand RP, St George-Hyslop P, Fraser PE. Defective membrane interactions of familial Parkinson's disease mutant A30P alpha-synuclein. J Mol Biol. 2002; 315:799-807.
• [46]Um JW, Im E, Lee HJ, Min B, Yoo L, Yoo J et al.. Parkin directly modulates 26S proteasome activity. J Neurosci. 2010; 30:11805-11814.
• [47]Ishii A, Nonaka T, Taniguchi S, Saito T, Arai T, Mann D et al.. Casein kinase 2 is the major enzyme in brain that phosphorylates Ser129 of human alpha-synuclein: Implication for alpha-synucleinopathies. FEBS Lett. 2007; 581:4711-4717.
• [48]Visanji NP, Wislet-Gendebien S, Oschipok LW, Zhang G, Aubert I, Fraser PE et al.. Effect of Ser-129 phosphorylation on interaction of alpha-synuclein with synaptic and cellular membranes. J Biol Chem. 2011; 286:35863-35873.
• [49]Waxman EA, Giasson BI. Specificity and regulation of casein kinase-mediated phosphorylation of alpha-synuclein. J Neuropathol Exp Neurol. 2008; 67:402-416.
• [50]Machiya Y, Hara S, Arawaka S, Fukushima S, Sato H, Sakamoto M et al.. Phosphorylated alpha-synuclein at Ser-129 is targeted to the proteasome pathway in a ubiquitin-independent manner. J Biol Chem. 2010; 285:40732-40744.
• [51]Chen L, Feany MB. Alpha-synuclein phosphorylation controls neurotoxicity and inclusion formation in a Drosophila model of Parkinson disease. Nat Neurosci. 2005; 8:657-663.
• [52]Kuwahara T, Tonegawa R, Ito G, Mitani S, Iwatsubo T. Phosphorylation of alpha-synuclein protein at Ser-129 reduces neuronal dysfunction by lowering its membrane binding property in Caenorhabditis elegans. J Biol Chem. 2012; 287:7098-7109.
• [53]AzeredodaSilveira S, Schneider BL, Cifuentes-Diaz C, Sage D, Abbas-Terki T, Iwatsubo T et al.. Phosphorylation does not prompt, nor prevent, the formation of alpha-synuclein toxic species in a rat model of Parkinson's disease. Hum Mol Genet. 2009; 18:872-887.
• [54]Gorbatyuk OS, Li S, Sullivan LF, Chen W, Kondrikova G, Manfredsson FP et al.. The phosphorylation state of Ser-129 in human alpha-synuclein determines neurodegeneration in a rat model of Parkinson disease. Proc Natl Acad Sci U S A. 2008; 105:763-768.
• [55]McFarland NR, Fan Z, Xu K, Schwarzschild MA, Feany MB, Hyman BT et al.. Alpha-synuclein S129 phosphorylation mutants do not alter nigrostriatal toxicity in a rat model of Parkinson disease. J Neuropathol Exp Neurol. 2009; 68:515-524.
• [56]Febbraro F, Sahin G, Farran A, Soares S, Jensen PH, Kirik D et al.. Ser129D mutant alpha-synuclein induces earlier motor dysfunction while S129A results in distinctive pathology in a rat model of Parkinson’s disease. Neurobiol Dis. 2013; 56:47-58.
• [57]Sato H, Arawaka S, Hara S, Fukushima S, Koga K, Koyama S et al.. Authentically phosphorylated alpha-synuclein at Ser129 accelerates neurodegeneration in a rat model of familial Parkinson’s disease. J Neurosci. 2011; 31:16884-16894.
• [58]Van der Perren A, Toelen J, Casteels C, Macchi F, Van Rompuy A-S, Sarre S et al.. Longitudinal follow-up and characterization of a robust rat model for Parkinson’s disease based on overexpression of alpha-synuclein with adeno-associated viral vectors. Neurobiol Aging. 2015; 36:1543-1558.
• [59]Gao G, Vandenberghe LH, Wilson JM. New recombinant serotypes of AAV vectors. Curr Gene Ther. 2005; 5:285-297.
• [60]Baekelandt V, Claeys A, Eggermont K, Lauwers E, De Strooper B, Nuttin B et al.. Characterization of lentiviral vector-mediated gene transfer in adult mouse brain. Hum Gene Ther. 2002; 13:841-853.
• [61]Schmitz C, Hof PR. Design-based stereology in neuroscience. Neuroscience. 2005; 130:813-831.
• [62]Osorio L, Gijsbers R, Oliveras-Salva M, Michiels A, Debyser Z, Van den Haute C et al.. Viral vectors expressing a single microRNA-based short-hairpin RNA result in potent gene silencing in vitro and in vivo. J Biotechnol. 2014; 169:71-81.