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mRNA analysis of α- and β-synemin and study of protein involving with α-synemin in brain

脑内α-和β-synemin mRNA分析及α-synemin相关蛋白研究

基本信息

批准号：
18590925
负责人：
MIZUNO Yuji
金额：
$ 2.43万
依托单位：
Gunma University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2006
资助国家：
日本
起止时间：
2006 至 2007
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-18590925/
关键词：
β-synemin desmuslin intermediate filament muscular dystrophy dystrobrevin beta-svnemin

项目摘要

Synemin is a muscle intermediate filament protein that was originally identified in birds. Recently, the human α- and β-synemin orthologues have been cloned, the latter of which was previously termed human desmuslin. Both human synemin isoforms derive from the same gene as a result of differential splicing between exons 4 and 5 such that the β-synemin protein is 312 amino acids shorter at its C-terminus. Both β-synemin mRNA and protein are highly expressed in skeletal and cardiac muscles, while northern blot analysis shows weak doublet in brain, indicating that there are two different types of synemin present in brain on the basis of mRNA level. We consider that β-synemin plays a role in maintaining the cytoskeletal structure in skeletal muscle, while α-synemin is also involving to role of the morphological maintenance of neuron and glia. We think that it is of great importance to elucidate the proteins involving α-synemin in brain. Therefore, we decided to examine human brain cDNA library to find a new sequence that interacts with the specific region to α-synemin, 312 amino acids sequences, using Matchmaker Two-hybrid system 3.Last year, we examined the expression of α- and β-synemin mRNA and found that both synemin mRNAs are expressed in the large neurons of dorsal side of pons and midbrain using in situ hybridization. This year, we further recognized that both synemin mRNA and its protein are co-localized in the same large neurons by in situ hybridization and immunohistochemical analysis.

Synemin是一种肌肉中间丝蛋白，最初在鸟类中发现。最近，人α-和β-突触蛋白直向同源物已被克隆，后者以前被称为人desmuslin。由于外显子4和5之间的差异剪接，两种人突触素同种型均源自相同基因，使得β-突触素蛋白质在其C末端短312个氨基酸。β-synemin mRNA和蛋白在骨骼肌和心肌中均高表达，而北方印迹分析显示脑中存在弱的双联体，表明在mRNA水平上脑中存在两种不同类型的synemin。我们认为β-synemin在维持骨骼肌细胞骨架结构方面起作用，而α-synemin也参与了神经元和胶质细胞的形态维持作用。我们认为，阐明脑内α-synemin的相关蛋白具有重要意义。因此，我们决定利用Matchmaker双杂交系统3，从人脑cDNA文库中寻找与α-synemin特异性区域312个氨基酸序列相互作用的新序列。去年，我们检测了α-synemin和β-synemin mRNA的表达，发现这两种synemin mRNA在脑桥背侧和中脑的大神经元中均有表达。今年，我们通过原位杂交和免疫组织化学分析进一步认识到synemin mRNA及其蛋白质在同一大神经元中共定位。