title: Alpha-synuclein immunoisolation of glial inclusions from multiple system atrophy brain tissue reveals multiprotein components.
abstract: Immunohistochemical studies have shown that oligodendroglial inclusions in multiple system atrophy contain alpha-synuclein, a synaptic protein also found in Lewy bodies in Parkinson's disease. We have now used density gradient enrichment and an anti-alpha-synuclein immunomagnetic technique to isolate pure and morphologically intact oligodendroglial inclusions from brain white matter of patients dying with multiple system atrophy. Filamentous inclusion structures were obtained only from multiple system atrophy tissue, but not from normal brain tissues, or from multiple system atrophy tissue processed without anti-alpha-synuclein antibody. We confirmed the purity and morphology of isolated inclusions by electron microscopy. The inclusions comprised multiple protein bands after separation by polyacrylamide gel electrophoresis. Immunoblotting demonstrated that these proteins included alpha-synuclein, alphaB-crystallin, tubulins, ubiquitin, and prominent, possibly truncated alpha-synuclein species as high-molecular-weight aggregates. Our study provides the first biochemical evidence that oligodendroglial inclusion filaments consist of multiple protein components, suggesting that these inclusions may form as a result of multiprotein interactions with alpha-synuclein.
authors.affiliation: Department of Human Physiology and Centre for Neuroscience, Flinders University, Bedford Park, South Australia.
authors.forename: W P
authors.initials: WP
authors.lastname: Gai
authors.forename: J H
authors.initials: JH
authors.lastname: Power
authors.forename: P C
authors.initials: PC
authors.lastname: Blumbergs
authors.forename: J G
authors.initials: JG
authors.lastname: Culvenor
authors.forename: P H
authors.initials: PH
authors.lastname: Jensen
mesh_terms: D000368:Aged; D019921:Biotinylation; D001921:Brain / Q000648:ultrastructure*; D002458:Cell Fractionation / Q000379:methods*; D002499:Centrifugation, Density Gradient; D003459:Crystallins / Q000032:analysis; D005260:Female; D006801:Humans; D015151:Immunoblotting; D018189:Immunomagnetic Separation; D002479:Inclusion Bodies / Q000737:chemistry / Q000648:ultrastructure*; D008854:Microscopy, Electron; D008875:Middle Aged; D019578:Multiple System Atrophy / Q000473:pathology*; D009419:Nerve Tissue Proteins / Q000032:analysis / Q000276:immunology*; D009457:Neuroglia / Q000648:ultrastructure*; D051843:Synucleins; D014404:Tubulin / Q000032:analysis; D014452:Ubiquitins / Q000032:analysis; D051844:alpha-Synuclein
chemical_list: D003459:Crystallins; D009419:Nerve Tissue Proteins; C497604:SNCA protein, human; D051843:Synucleins; D014404:Tubulin; D014452:Ubiquitins; D051844:alpha-Synuclein