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

Thomas M. Wishart, Timothy M. Rooney, Douglas J. Lamont, Ann K. Wright, A. Jennifer Morton, Mandy Jackson, Marc Freeman, Thomas H. Gillingwater

Research output: Contribution to journalArticle

32 Citations (Scopus)

Abstract

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.

Original languageEnglish (US)
Article numbere1002936
JournalPLoS Genetics
Volume8
Issue number8
DOIs
StatePublished - Aug 2012
Externally publishedYes

Fingerprint

proteomics
synapse
molecular genetics
Proteomics
Synapses
Molecular Biology
synaptic transmission
protein
Synaptic Transmission
proteins
neurons
brain
mutation
Spinocerebellar Ataxias
Neurons
calcium-binding proteins
mutants
Synaptic Potentials
Mutation
molecular chaperones

ASJC Scopus subject areas

  • Genetics
  • Molecular Biology
  • Ecology, Evolution, Behavior and Systematics
  • Cancer Research
  • Genetics(clinical)

Cite this

Wishart, T. M., Rooney, T. M., Lamont, D. J., Wright, A. K., Morton, A. J., Jackson, M., ... Gillingwater, T. H. (2012). Combining Comparative Proteomics and Molecular Genetics Uncovers Regulators of Synaptic and Axonal Stability and Degeneration In Vivo. PLoS Genetics, 8(8), [e1002936]. https://doi.org/10.1371/journal.pgen.1002936

Combining Comparative Proteomics and Molecular Genetics Uncovers Regulators of Synaptic and Axonal Stability and Degeneration In Vivo. / Wishart, Thomas M.; Rooney, Timothy M.; Lamont, Douglas J.; Wright, Ann K.; Morton, A. Jennifer; Jackson, Mandy; Freeman, Marc; Gillingwater, Thomas H.

In: PLoS Genetics, Vol. 8, No. 8, e1002936, 08.2012.

Research output: Contribution to journalArticle

Wishart, Thomas M. ; Rooney, Timothy M. ; Lamont, Douglas J. ; Wright, Ann K. ; Morton, A. Jennifer ; Jackson, Mandy ; Freeman, Marc ; Gillingwater, Thomas H. / Combining Comparative Proteomics and Molecular Genetics Uncovers Regulators of Synaptic and Axonal Stability and Degeneration In Vivo. In: PLoS Genetics. 2012 ; Vol. 8, No. 8.
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