原代培养脊髓神经元线状溶酶体与细胞骨架蛋白-纽蛋白关系的研究  

RELATIONSHIP BETWEEN VINCULIN THE CYTOSKELETAL PROTEIN ANDNEMATOLYSOSOMES IN PRIMARY CULTURED NEURONS OF SPINAL CORD

作　　者：徐延豪[1] 荣明[1] 石玉秀[1] 

机构地区：[1]中国医科大学组织学与胚胎学教研室,沈阳110001

出　　处：《中国组织化学与细胞化学杂志》2004年第4期494-498,共5页Chinese Journal of Histochemistry and Cytochemistry

基　　金：国家自然科学基金资助课题(39970383)

摘　　要：目的研究原代培养脊髓神经元线状溶酶体(nematolysosome)的形成与分布及其与细胞骨架蛋白纽蛋白(vinculin)的关系。方法用细胞松弛素D(cytochalasinD,CD)及佛波醇酯(phorbolmyristateacetate,PMA)处理原代培养脊髓神经元,用免疫荧光双标记纽蛋白及组织蛋白酶D(cathepsinD)、酸性磷酸酶(ACPase)、电镜细胞化学及共焦激光扫描显微镜方法研究线状溶酶体与纽蛋白的关系。结果在正常对照组神经元,组织蛋白酶D(标记溶酶体)与纽蛋白分布于胞质及突起内;在CD及PMA处理神经元,纽蛋白及组织蛋白酶D的分布呈向心性移动,但集聚的部位不同;电镜酶细胞化学方法显示CD组及PMA组神经元内线状溶酶体均增多。结论组织蛋白酶D及纽蛋白在培养脊髓神经元内协同分布,CD及PMA均可引起二者分布的变化,提示纽蛋白可通过增强细胞内吞体/溶酶体系统活动而使线状溶酶体增加,也可通过促进丝状肌动蛋白聚合而影响线状溶酶体的形成及运动。? Objective To study the function of a cytoskeletal protein——vinculin——in cellular activity and its effects on the formation and function of nematolysosomes. Methods The spinal cord of the newborn rat was used for primary cell culture, and the cells were treated by cytochalasin D (CD group) and PMA (PMA group).Then histochemical, electron microscopic, enzyme histo cytochemical and confocal laser scanning microscopic methods were employed to study the distribution of vinculin, cathepsin Dand nematolysosomes and the relationship between nematolysosomes and vinculin. Results Both CD and PMA cause changes not only in the distribution and expression of vinculinand cathepsin D, but also in the formation and distribution of nematolysosomes. The nematolysosomes increased more significantly in the neurons of the PMA group than in those of the CD group. Conclusion The expression of vinculin is related to distribution of Cathepsin D. Vinculin might increase the formation of nematolysosomes by promoting the activity of the endosomal/lysosomal system. Vinculin might also stimulate the transportation of nematolysosomes by initiating the polymerization of F actin.〔

关 键 词：线状溶酶体 纽蛋白 组织蛋白酶D 激光共焦扫描显微镜 

分 类 号：R322.812[医药卫生—人体解剖和组织胚胎学]