细丝蛋白A影响大鼠血睾屏障形成初期肌动蛋白丝的排布
Filamin A affects F-actin arrangement during early formation of rat blood-testis barrier
目的：基于肌动蛋白丝（F-actin）在血睾屏障（BTB）形成与变化过程中的作用，初步探讨细丝蛋白A（filamin A）在大鼠血睾屏障形成初期对F-actin排布的影响。方法：Western印迹检测filamin A在大鼠睾丸、支持细胞及精子中的表达；免疫荧光染色观察filaminA与F-actin在20 d龄大鼠生精上皮中的共定位；体外无血清体系分离培养大鼠睾丸支持细胞，待细胞连接形成后通过siRNA干扰沉默filamin A的表达，鬼笔环肽染色观察F-actin的分布定位；睾丸内siRNA注射，观察filamin A对血睾屏障形成时F-actin分布的影响。结果：filamin A在20 d龄大鼠主要由支持细胞表达，体外siRNA沉默filamin A的效率可达70%以上，在体外支持细胞连接形成及体内屏障形成过程中，filamin A沉默组的F-actin与对照组相比排列紊乱、无序。结论：filamin A参与调节大鼠血睾屏障形成初期F-actin的排布。
Objective: To investigate the role of filamin A in arrangement of F-actin during initial formation of rat blood-testis barrier (BTB). Methods: The expression of filamin A in rat testis, Sertoli cells and sperm were detected by Western blot. Co-localization of filamin A and F-actin was observed by immunofluorescence staining. Rat Sertoli cells was isolated and cultured in vitro in a serum free system. The expression of filamin A was silenced using siRNA duplexes after well cell junctions were formed. The localization of F-actin was observed by Cy3-phylloidin staining. Intro-testicular injection of siRNA mix was used to silence filamin A in vivo and frozen sections were prepared to detect F-actin arrangement after filamin A silencing. Result: filamin A was expressed in 20-day-old rat mainly by Sertoli cells. The silencing effect of filamin A in vitro can be over 70% . The silencing of filamin A both in vitro and in vivo can lead to rearrangement and mislocalization of F-actin. Conclusion: filamin A participate in the localization of F-actin during the initial formation of rat BTB.
张丽雁、宿文辉
细胞生物学分子生物学
血睾屏障肌动蛋白丝细丝蛋白
blood-testis barrierF-actinfilamin
张丽雁,宿文辉.细丝蛋白A影响大鼠血睾屏障形成初期肌动蛋白丝的排布[EB/OL].(2012-06-18)[2025-05-01].http://www.paper.edu.cn/releasepaper/content/201206-252.点此复制
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