仔猪睾丸支持细胞的增殖与发育  

Proliferation and Development of Piglet Sertoli Cells

作　　者：周银涛[1] 王鲜忠[1] 周玉兰[1] 赵伯川[1] 陈永军[1] 张家骅[1] 

机构地区：[1]西南大学动物科技学院重庆市牧草与草食家畜重点实验室,重庆400716

出　　处：《农业生物技术学报》2011年第1期128-134,共7页Journal of Agricultural Biotechnology

基　　金：国家自然科学基金项目(No.31072183);中央高校基本科研业务费专项项目(No.XDJK2009B035)资助

摘　　要：为了阐明支持细胞增殖和分化能力对精子生成的影响,本实验以7、21和35d仔猪为材料,利用细胞直接计数、免疫组织化学、流式细胞术和免疫印迹分析了仔猪睾丸支持细胞、生精细胞数量和增殖能力变化以及支持细胞的成熟分化状况。结果:(1)从7～21d,曲细精管中支持细胞与其它细胞之间的界线更加明显;从7～35d,每克睾丸中间质细胞和支持细胞的数量在21d时达到高峰(P<0.05),而生精细胞的数量则一直增加(P<0.05),但并没有在曲细精管中发现圆形精子;(2)从7～35d,支持细胞和间质细胞都能表达GATA-4,支持细胞中GATA-4的累积光密度(IOD)在21d时最高(P<0.05),而间质细胞中的IOD在21d和35d之间没有明显差异(P>0.05);(3)从7～35d,整个睾丸细胞都有增殖能力,睾丸细胞的细胞增殖指数(PI)和S期细胞指数(SPF)在21d时最高(P<0.05);三种细胞在这一时期都能表达PCNA,三种细胞PCNA的IOD值和睾丸中PCNA的表达水平在21d时最高(P<0.05);(4)从7～35d,Connexin 43(Cx43)主要表达于支持细胞的细胞质,支持细胞和间质细胞中的IODs在21d时最高(P<0.05)。结果表明,尽管在出生后1个月左右仔猪睾丸支持细胞有较强的增殖能力,但由于这一时期的支持细胞没有分化成熟,因此,不能支持生精细胞的进一步发育。This study was performed to explain effect of Sertoli cells on spermatogenesis. Testes of porcine from 7 d-old,21 d-old,35 d-old were used,and the methods including cell count,immunohistochemistry,flow cytometry and western blot were used. The results were as follows：（1）from 7 d to 21 d,the boundary between Sertoli cell and other cells in seminiferous tubules was more significant. From 7 d to 35 d,the number of Sertoli cells and Leydig cells and of per gram of testis increased（P0.05）,and the number of both cells were the most at 21 d（P0.05）. After that period,the number of both cells of per gram of testis began to decrease（P0.05）. However,the number of germ cells of per gram of testis continued to increase（P0.05）,but round spermatids could not be found in the seminiferous tubule. （2） form 7 d to 35 d,both of Sertoli cells and Leydig cells could express GATA-4,and the integrated optical density（IOD） of GATA-4 of Sertoli cells was the most at 21 d（P0.05）,but that of Leydig cells between at 21 d and 35 d had no difference（P0.05）. （3） form 7 d to 35 d,total testical cells had proliferation capacities,while PI and SPF in testes were the most at 21 d（P0.05）. During the periods,three types of cells（Sertoli cells,Leydig cells and germ cells） could express PCNA,and the IODs of PCNA of three cells were the most at 21 d（P0.05）. the expression of PCNA（Proliferating Cell Nuclear Antigen） in the testes was the most at 21 d（P0.05）. （4） form 7 d to 35 d,Connexin 43（Cx43） was mainly expressed in cytoplasm of Sertoli cells,and the IODs of Cx43 of Sertoli cells and Leydig cells were the most at 21 d（P0.05）. The results infer that it is an important proliferation peroid form 7 d to 35 d,but these immature sertoli cells could not support spermatogenesis.

关 键 词：仔猪 睾丸 支持细胞 生精细胞 细胞增殖 分化 

分 类 号：S865.12[农业科学—野生动物驯养]