Biochemical study of glial cytoplasmic inclusions in multiple system atrophy brains

胶质细胞胞浆包涵体（GCI）是多系统萎缩（MSA）的主要病理特征，主要存在于少突胶质细胞中。本研究尝试了GCI的几种检测方法，包括免疫组化法，发现Gallyas染色法是GCI的最佳检测方法，并利用Gallyas染色法建立了GCI的部分纯化方法，进一步电镜观察到了Triton不溶性GCI富集组分中的纤维状GCI，Gallyas银染色法也观察到了纤维状GCI。然后我们对富含GCI的级分进行了2D凝胶分析。该分析揭示了蛋白质的量增加，其显示MW 22 kD和p17.0-7.6对应于α B-晶体蛋白。用抗α B-晶体蛋白抗体和Gallyas银染色对该级分的2D凝胶的免疫印迹进行染色。抗α B-晶体蛋白抗体与对应于α B-晶体蛋白的GCI富集级分中增加的斑点反应。Gallyas银染色也染色这些斑点，表明α B-晶体蛋白是Gallyas反应蛋白之一。这些结果表明，GCI的主要成分之一是α-B-晶体蛋白.此外，抗羧基甲基转移酶抗体染色GCI，表明这些异常蛋白在MSA脑中发生了外消旋化.我们获得了几种针对遗传性神经疾病基因产物的抗体，以研究这些蛋白与GCI的关系.这些抗体检测三核苷酸疾病中的异常基因产物.

Glial cytoplamic inclusions (GCI) which appear in oligodendroglial cells are the main pathological feature of multiple system atrophy (MSA). In this study, to determine the main constituent of GCI we tried several assay methods of GCI including immunohistochemical methods and found that Gallyas staining is the best assay method of GCI.Using Gallyas staining we developed the partial purification method of GCI.Furthermore electron micrograph showed the fibrous GCI in the Triton insoluble GCI-enriched fraction which was also observed with Gallyas silver staining. Then we performed 2D gel analysis of the GCI-rich fraction. This analysis revealed the increased amount of the protein which showed MW22kD and p17.0-7.6 corresponding to alpha B-Crystallin. Immunoblots of 2D gel of that fraction were stained with anti-alpha B-Crystallin antibodies and with Gallyas silver staining. Anti-alpha B-Crystallin antibodies reacted with the spots which increased in GCI-enriched fraction corresponding to alpha B-Crystallin. Gallyas silver staining also stained these spots, suggesting that alpha B-Crystallin is one of the Gallyas reactive proteins. These results suggests that one of the main constituent of GCI is alpha B-Crystallin.Furthermore anti-carboxyl methtltransferase antibodies stained GCI,suggesting that the racemization of those abnormal proteins occured in MSA brain.We got several antibodies against the gene products of genetic neurological disorders to study the relationship between these protein and GCI.These antibodies detect abnormal gene products in trinucleotide diseases.