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αB-Crystallin is a major component of glial cytoplasmic inclusions in multiple system atrophy

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Abstract

Multiple system atrophy (MSA) is characterized by the formation of oligodendroglial cytoplasmic inclusions (GCIs) consisting of α-synuclein filaments. αB-crystallin, a small chaperone protein that binds to unfolded proteins and inhibits aggregation, has been documented in GCIs. We investigated the relative abundance and speciation of αB-crystallin in GCIs in MSA brains. We also examined the influence of αB-crystallin on the formation of cytoplasmic inclusions in cultured glial cells. Immunohistochemistry and confocal microscopy revealed αB-crystallin is a prominent component of GCIs, more abundant than in Lewy bodies in Lewy body dementia. One- and two-dimensional gel electrophoresis and mass spectrometric analysis of GCIs immunopurified from MSA brains indicated that αB-crystallin is a major protein component with multiple post-translationally modified species. In cultured C6 glioma cells treated with the proteasomal inhibitor, lactacystin, to induce accumulation of ubiquitinated proteins, a subset of cells showed increased cytoplasmic staining for αB-crystallin. Proteasomeinhibited cells transfected with GFP-tagged α-synuclein resulted in ubiquitin- and αB-crystallin-positive aggregates resembling GCIs in MSA brains. Our results indicate that αB-crystallin is a major chaperone in MSA, and suggest a role of the protein in the formation of inclusion bodies in glial cells.

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Abbreviations

MSA:

multiple system atrophy

PD:

Parkinson’s disease

DLB:

Dementia with Lewy bodies

LB:

Lewy body

GCI:

glial cytoplasmic inclusion

PTM:

posttranslational modification

ESI:

electrospray ionisation

MS:

mass spectrometry

SDS-PAGE:

sodium dodecyl sulphate-polyacrylamide gel electrophoresis

CHAPS:

3-[(3-Cholamidopropyl) dimethyl-ammonio]-1-propanesulfonate

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Authors and Affiliations

  1. Department of Human Physiology, Flinders University, 5001, Adelaide, Australia

    D. L. Pountney, T. Chataway, F. Chegini, P. C. Blumbergs & W. P. Gai

  2. Departments of Chemistry and Biological Sciences, University of Wollongong, Australia

    T. M. Treweek

  3. Prince of Wales Medical Research Institute, Sydney, Australia

    Y. Huang

  4. Department of Neuropathology, Institute of Medical and Veterinary Science, Adelaide, Australia

    F. Chegini

  5. School of Biomedical Sciences, University of New South Wales, Australia

    M. J Raftery

Authors
  1. D. L. Pountney
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  2. T. M. Treweek
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  3. T. Chataway
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  4. Y. Huang
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  5. F. Chegini
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  6. P. C. Blumbergs
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  7. M. J Raftery
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  8. W. P. Gai
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Correspondence to W. P. Gai.

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Pountney, D.L., Treweek, T.M., Chataway, T. et al. αB-Crystallin is a major component of glial cytoplasmic inclusions in multiple system atrophy. neurotox res 7, 77–85 (2005). https://doi.org/10.1007/BF03033778

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  • DOI: https://doi.org/10.1007/BF03033778

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