Nogo-66受体分子在体外培养的星形胶质细胞中的表达  

Expression of Nogo-66 receptor in primary cultured astrocytes

作　　者：孙芳[1] 金卫林[1,2] 龙梅 鞠躬[1,2] 

机构地区：[1]第四军医大学神经科学研究所 [2]上海交通大学医学院,上海200030

出　　处：《神经科学通报》2005年第4期273-277,共5页Neuroscience Bulletin

基　　金：supported by a grant from NSFC/RGC(No.30218003);a grant from 973 National Key Basic Research Program of China(No.2003CB515300).

摘　　要：目的检测Nogo-66受体(NgR)在体外培养的大鼠星形胶质细胞中的表达。方法利用RT-PCR、West-ern Blot和间接免疫荧光染色技术,检测原代培养并纯化的星形胶质细胞中NgRm RNA和蛋白分子的表达。结果利用RT-PCR技术,从星形胶质细胞总RNA中扩增出特异的NgR条带;利用Western Blot技术,从星形胶质细胞抽提物中检测到64kD特异性的NgR反应条带;间接免疫荧光染色和Confocal显微镜进一步显示,星形胶质细胞中的NgR免疫反应物主要位于细胞内。同时,来源于大鼠的C6胶质瘤细胞也证明表达NgR蛋白。结论体外培养的星形胶质细胞中表达NgR分子,这为证明NgR在体内胶质细胞中的表达提供了更直接的证据。Objective To investigate the expression of Nogo-66 receptor （NgR） in primary cultured rat astrocytes.Methods RT-PCR and Western blot were applied to verify mRNA and protein expression of NgR in primary cultured and purified astrocytes. Indirect immunofluorensence and confocal microscopic technique were used to study the distribution of NgR. Results Specific NgR cDNA product could be amplified from the total RNA of primary cultured astrocytes by RTPCR ; Western blot of the extracts of astrocytes demonstrated a specific NgR band at about 64 kD. Indirect immunofluorescence and confocal scanning further revealed the intracellular localization of NgR protein in astrocytes. Simultaneously,the NgR protein was detected in C6 rat glioma cells by western blot and immunofluoresence staining. Conclusion NgR is expressed in primary cultured astrocytes,which offers strong and direct support for the expression of NgR in astrocytes in uiuo.

关 键 词：Nogo-66受体 星形胶质细胞 体外 表达 

分 类 号：Q813.11[生物学—生物工程]