仔猪睾丸支持细胞的增殖与发育

摘要

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(P＜0.05), and the number of both cells were the most at 21 d (P＜0.05).After that period, the number of both cells of per gram of testis began to decrease (P＜0.05).However,the number of germ cells of per gram of testis continued to increase (P＜0.05), 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(P＜0.05), but that of Leydig cells between at 21 d and 35 d had no difference (P＞0.05).(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 (P＜0.05).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(P＜0.05).the expression of PCNA(Proliferating Cell Nuclear Antigen) in the testes was the most at 21 d(P＜0.05).(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 (P＜0.05).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.%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35 d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7～21 d,曲细精管中支持细胞与其它细胞之间的界线更加明显;从7～35 d,每克睾丸中间质细胞和支持细胞的数量在21 d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7～35 d,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21 d时最高(P<0.05),而间质细胞中的IOD在21 d和35 d之间没有明显差异(P>0.05);(3)从7～35 d,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21 d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21 d时最高(P<0.05);(4)从7～35 d,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21 d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.