Combining Comparative Proteomics and Molecular Genetics Uncovers Regulators of Synaptic and Axonal Stability and Degeneration

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Degeneration of synaptic and axonal compartments of neurons is an early event contributing to the pathogenesis of many neurodegenerative diseases, but the underlying molecular mechanisms remain unclear. Here, we demonstrate the effectiveness of a novel “top-down” approach for identifying proteins and functional pathways regulating neurodegeneration in distal compartments of neurons. A series of comparative quantitative proteomic screens on synapse-enriched fractions isolated from the mouse brain following injury identified dynamic perturbations occurring within the proteome during both initiation and onset phases of degeneration. In silico analyses highlighted significant clustering of proteins contributing to functional pathways regulating synaptic transmission and neurite development. Molecular markers of degeneration were conserved in injury and disease, with comparable responses observed in synapse-enriched fractions isolated from mouse models of Huntington's disease (HD) and spinocerebellar ataxia type 5. An initial screen targeting thirteen degeneration-associated proteins using mutant Drosophila lines revealed six potential regulators of synaptic and axonal degeneration in vivo. Mutations in CALB2, ROCK2, DNAJC5/CSP, and HIBCH partially delayed injury-induced neurodegeneration. Conversely, mutations in DNAJC6 and ALDHA1 led to spontaneous degeneration of distal axons and synapses. A more detailed genetic analysis of DNAJC5/CSP mutants confirmed that loss of DNAJC5/CSP was neuroprotective, robustly delaying degeneration in axonal and synaptic compartments. Our study has identified conserved molecular responses occurring within synapse-enriched fractions of the mouse brain during the early stages of neurodegeneration, focused on functional networks modulating synaptic transmission and incorporating molecular chaperones, cytoskeletal modifiers, and calcium-binding proteins. We propose that the proteins and functional pathways identified in the current study represent attractive targets for developing therapeutics aimed at modulating synaptic and axonal stability and neurodegeneration in vivo.

Vyšlo v časopise: Combining Comparative Proteomics and Molecular Genetics Uncovers Regulators of Synaptic and Axonal Stability and Degeneration. PLoS Genet 8(8): e32767. doi:10.1371/journal.pgen.1002936
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.pgen.1002936

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Zdroje

1. WishartTM, ParsonSH, GillingwaterTH (2006) Synaptic vulnerability in neurodegenerative disease. J Neuropathol Exp Neurol 65 : 733–739.

2. SelkoeDJ (2002) Alzheimer's disease is a synaptic failure. Science 298 : 789–791.

3. Schulz-SchaefferWJ (2010) The synaptic pathology of alpha-synuclein aggregation in dementia with Lewy bodies, Parkinson's disease and Parkinson's disease dementia. Acta Neuropathol 20 : 131–143.

4. MurphyKP, CarterRJ, LioneLA, MangiariniL, MahalA, et al. (2000) Abnormal synaptic plasticity and impaired spatial cognition in mice transgenic for exon 1 of the human Huntington's disease mutation. J Neurosci 20 : 5115–23.

5. PerkinsEM, ClarksonYL, SabatierN, LonghurstDM, MillwardCP, et al. (2010) Loss of beta-III spectrin leads to Purkinje cell dysfunction recapitulating the behavior and neuropathology of spinocerebellar ataxia type 5 in humans. J Neurosci 30 : 4857–67.

6. CunninghamC, DeaconR, WellsH, BocheD, WatersS, et al. (2003) Synaptic changes characterize early behavioural signs in the ME7 model of murine prion disease. Eur J Neurosci 17 : 2147–2155.

7. KielarC, WishartTM, PalmerA, DianichS, MacauleySL, et al. (2009) Molecular correlates of axonal and synaptic pathology in mouse models of Batten disease. Hum Mol Genet 18 : 4066–80.

8. FischerLR, CulverDG, TennantP, DavisAA, WangM, et al. (2004) Amyotrophic lateral sclerosis is a distal axonopathy: evidence in mice and man. Exp Neurol 185 : 232–240.

9. MurrayLM, ComleyLH, ThomsonD, ParkinsonN, TalbotK, et al. (2008) Selective vulnerability of motor neurons and dissociation of pre -⁠ and post-synaptic pathology at the neuromuscular junction in mouse models of spinal muscular atrophy. Hum Mol Genet 17 : 949–62.

10. ColemanP, FederoffH, KurlanR (2004) A focus on the synapse for neuroprotection in Alzheimer disease and other dementias. Neurology 63 : 1155–1162.

11. BurgoyneRD, MorganA (2011) Chaperoning the SNAREs: a role in preventing neurodegeneration? Nat Cell Biol 13 : 8–9.

12. LunnER, PerryVH, BrownMC, RosenH, GordonS (1989) Absence of wallerian degeneration does not hinder regeneration in peripheral nerve. Eur J Neurosci 1 : 27–33.

13. MackTGA, ReinerM, BeirowskiB, MiW, EmanuelliM, et al. (2001) Wallerain degeneration of injured axons and synapses is delayed by a Ube4b/Nmnat chimeric gene. Nat Neurosci 4 : 1199–1206.

14. GillingwaterTH, InghamCA, ParryKE, WrightAK, HaleyJE, et al. (2006) Delayed Synaptic Degeneration in the CNS of Wlds Mice After Cortical Lesion. Brain 129 : 1546–1556.

15. Fernández-ChacónR, WölfelM, NishimuneH, TabaresL, SchmitzF, et al. (2004) The synaptic vesicle protein CSP alpha prevents presynaptic degeneration. Neuron 42 : 237–251.

16. ChandraS, GallardoG, Fernández-ChacónR, SchlüterOM, SüdhofTC (2005) Alpha-synuclein cooperates with CSPalpha in preventing neurodegeneration. Cell 123 : 383–396.

17. BurréJ, SharmaM, TsetsenisT, BuchmanV, EthertonMR, et al. (2010) Alpha-synuclein promotes SNARE-complex assembly in vivo and in vitro. Science 329 : 1663–1667.

18. WishartTM, PatersonJM, ShortDM, MeredithS, RobertsonKA, et al. (2007) Differential Proteomic Analysis of Synaptic Proteins Identifies Potential Cellular Targets and Protein Mediators of Synaptic Neuroprotection Conferred by the Slow Wallerian Degeneration (Wlds) Gene. Mol Cell Proteom 6 : 1318–1330.

19. WrightAK, WishartTM, InghamCA, GillingwaterTH (2010) Synaptic protection in the brain of WldS mice occurs independently of age but is sensitive to gene-dose. PLoS ONE 5: e15108 doi:10.1371/journal.pone.0015108.

20. MortonAJ, LaganMA, SkepperJN, DunnettSB (2000) Progressive formation of inclusions in the striatum and hippocampus of mice transgenic for the human Huntington's disease mutation. J Neurocytol 29 : 679–702.

21. MacDonaldJM, BeachMG, PorpigliaE, SheehanAE, WattsRJ, et al. (2006) The Drosophila cell corpse engulfment receptor Draper mediates glial clearance of severed axons. Neuron 50 : 869–881.

22. OhtsukaK, HataM (2000) Mammalian HSP40/DNAJ homologs: cloning of novel cDNAs and a proposal for their classification and nomenclature. Cell Stress Chaperones 5 : 98–112.

23. RogersJH (1987) Calretinin: A gene for a novel calcium-binding protein expressed principally in neurons. J Cell Biol 105 : 1343–1353.

24. RientoK, RidleyAJ (2003) Rocks: multifunctional kinases in cell behaviour. Nat Rev Mol Cell Biol 4 : 446–456.

25. GurdenH, SchiffmannSN, LemaireM, BöhmeGA, ParmentierM, SchurmansS (1998) Calretinin expression as a critical component in the control of dentate gyrus long-term potentiation induction in mice. Eur J Neurosci 10 : 3029–3033.

26. AbeK, YamaguchiS, SugiuraM, SaitoH (1999) The ethanol metabolite acetaldehyde inhibits the induction of long-term potentiation in the rat dentate gyrus in vivo. Br J Pharmacol 127 : 1805–1810.

27. MaycoxPR, LinkE, ReetzA, MorrisSA, JahnR (1992) Clathrin-coated vesicles in nervous tissue are involved primarily in synaptic vesicle recycling. J Cell Biol 118 : 1379–1388.

28. FotinA, ChengY, GrigorieffN, WalzT, HarrisonSC, et al. (2004) Structure of an auxilin-bound clathrin coat and its implications for the mechanism of uncoating. Nature 432 : 649–653.

29. SharmaM, BurréJ, SüdhofTC (2011) CSPα promotes SNARE-complex assembly by chaperoning SNAP-25 during synaptic activity. Nat Cell Biol 13 : 30–39.

30. ZhouZ, MengY, AsrarS, TodorovskiZ, JiaZ (2009) A critical role of Rho-kinase ROCK2 in the regulation of spine and synaptic function. Neuropharmacology 56 : 81–89.

31. LoupattyFJ, ClaytonPT, RuiterJP, OfmanR, IjlstL, et al. (2007) Mutations in the gene encoding 3-hydroxyisobutyryl-CoA hydrolase results in progressive infantile neurodegeneration. Am J Hum Genet 80 : 195–199.

32. WattsRJ, HoopferED, LuoL (2003) Axon pruning during Drosophila metamorphosis: evidence for local degeneration and requirement of the ubiquitin-proteasome system. Neuron 38 : 871–885.

33. NoskováL, StráneckýV, HartmannováH, PřistoupilováA, BarešováV, et al. (2011) Mutations in DNAJC5, Encoding Cysteine-String Protein Alpha, Cause Autosomal-Dominant Adult-Onset Neuronal Ceroid Lipofuscinosis. Am J Hum Genet 89 : 241–52.

34. SchmitzF, TabaresL, KhimichD, StrenzkeN, de la Villa-PoloP, et al. (2006) CSPalpha-deficiency causes massive and rapid photoreceptor degeneration. Proc Natl Acad Sci U S A 103 : 2926–2931.

35. WishartTM, HuangJP, MurrayLM, LamontDJ, MutsaersCA, et al. (2010) SMN deficiency disrupts brain development in a mouse model of severe spinal muscular atrophy. Hum Mol Genet 19 : 4216–4228.