纽蛋白在神经干细胞定向分化为神经元过程中的表达及其与辅肌动蛋白的关系  

EXPRESSION OF VINCULIN IN CULTURED CORTICAL STEM CELLS DURING DIRECTIONAL DIFFERENTIATION INTO NEURONS AND ITS RELATION WITHα-ACTININ

作　　者：徐延豪[1] 杜喆[1] 石玉秀[2] 

机构地区：[1]沈阳医学院,沈阳110034 [2]中国医科大学基础医学院组织胚胎学教研室,沈阳110001

出　　处：《中国组织化学与细胞化学杂志》2005年第5期582-587,共6页Chinese Journal of Histochemistry and Cytochemistry

基　　金：辽宁省教育厅科研攻关课题(202013132);辽宁省科技厅科技攻关课题(2003225007)

摘　　要：目的探讨纽蛋白(vinculin)在新生大鼠大脑皮层神经干细胞定向分化神经元过程中的表达变化及其与辅肌动蛋白(α-actinin)的关系。方法将神经干细胞分离、培养、分化及鉴定后,采用免疫组化技术对新生大鼠皮层神经干细胞向神经元定向分化过程中,纽蛋白的时空表达进行研究,并采用免疫荧光双标技术及共聚焦激光扫描显微术对神经元定向分化过程中,纽蛋白与辅肌动蛋白关系进行探讨,利用图像分析技术对不同时段分化的神经元中vinculin平均积分光密度进行定量测定。结果在神经干细胞定向分化为神经元的过程中,vinculin由核周淡染分布逐渐至在胞体与突起中密集均匀分布,随着突起伸展而不断地延伸。图像分析结果表示,随神经元的分化成熟,vinculin的表达量呈逐渐增加趋势。在共聚焦激光扫描显微镜下观察免疫荧光双标vinculin/α-actinin在分化早期胞体和突起内均有分布,可见核周和突起两者融合两蛋白共存。近成熟期,随突起生长,两蛋白渐伸入突起,直至末端共存。结论本研究揭示大脑皮质神经干细胞定向分化为神经元过程中,vinculin表达变化与神经元发育成熟呈正相关,并且vinculin与α-actinin共存。Objective To investigate the expression of directional differentiation into neurons and the relationship vinculin in cultured cortical stem cells during between vinculin and α-actinin. Methods We cultured, separated, differentiated and identified the cortex derived neural stem cells, and used the immunohistochemical technique to observe the distribution of vinculin in different stages of differentiated neurons. Meanwhile, by double-label immunofluorescent method and CLSM we explored the relationship - between it and α-actinin, and image analysis was used to measure the integrated optical density of vinculin in different stages of differentiated neurons. Results During the directional differentiation, vinculin was expressed slightly around the nucleus in the earlier stage and then intensively in cytoplasm and apophysis. Image analysis also showed that the expression of vinculin increased with the growth of neurons. Under CLSM we observed the distribution of vinculin and α-actinin in cell body and protuberance in the early stage and then the coexistence of the two albumen around nucleus and protuberance. In the process of maturity, with the growth of the protuberance, the two albumen went gradually in the protuberance until the bottoms coexisted. Conclusion These results suggest that the expression of vinculin is positively related to the maturity of neurons, and works in coordination with α-actinin in cultured cortical stem cells during directional differentiation into neurons.

关 键 词：纽蛋白 神经干细胞 定向分化 神经元 辅肌动蛋白 

分 类 号：R322.811[医药卫生—人体解剖和组织胚胎学]