摘要
目的Nogo-A在中枢神经系统的髓鞘和神经元中广泛表达,并且有多种亚分布形式。在以往研究的基础上,将通过检测Nogo-A/B(N-18)在大鼠脑组织中的亚细胞定位,进一步获得Nogo-A在细胞核中表达的有力证据。方法通过差速离心法和NP-40处理得到大鼠皮层和小脑样本的纯化细胞核亚组分,利用Westernblot来检测各亚细胞成分中Nogo-A的表达。通过免疫荧光双标染色,共聚焦显微镜观察Nogo-A在大鼠皮层和小脑组织中的亚细胞定位。结果WesternBlot在纯化细胞核中可检测到清晰的Nogo-A条带。免疫荧光染色显示胞核内有免疫阳性物质表达,并且和Hoechst复染的胞核着色有广泛重合。结论Nogo-A在大鼠皮层和小脑细胞核中有明确表达,并且很有可能以全长形式入核。
Objective Nogo-A is found widespread in both myelin and neurons of central nervous system in several ways of subeellular localization. Based on our previous research work, to get further evidence for intranuclear localization of Nogo-A by checking the subcellular localization of nogo-A/B in rat brain tissue. Methods Relatively pure nuclear fraction was got from rat cortex and cerebellum by differential centrifugation and NP-40. Then the expression of Nogo-A in the subcellular fractions were checked by Western blot. By double immunofluorescen, subcellular localization of Nogo-A was observed under confocal microscope. Result Clear Nogo-A band could be detected in purified nuclear fractions by Western blot. According to the immunofluorescen, positive signals were mainly found in the nucleus. Conclusion Nogo-A exists in nucleus of rat cortex and cerebellum and probably goes into the nucleus in the form of full length.
出处
《解剖学研究》
CAS
2009年第1期36-39,共4页
Anatomy Research
基金
NSFC/RGC联合科研基金(30218003)
