不同繁殖周期成年牦牛睾丸细胞外基质相关蛋白的分布比较

Distribution of extracellular matrix components in the adult yak testis during seasonal breeding cycle

应用Masson三色染色法、Gomori银浸染色法以及免疫组织化学SP法结合IPP统计分析法,比较了层黏连蛋白(LN)、Ⅳ型胶原蛋白(ColⅣ)和硫酸乙酰肝素糖蛋白(HSPG)在繁殖期和繁殖间期牦牛睾丸的分布及组织化学特征。结果,不同繁殖期牦牛睾丸间质胶原纤维及网状纤维均较为丰富;繁殖间期牦牛睾丸LN在支持细胞(sertoli cells)和生精细胞表达与繁殖期相近,在间质细胞(leydig cells)表达降低,但其平均吸光度检测无统计学差异(P>0.05);ColⅣ在不同繁殖期牦牛睾丸支持细胞和各级生精细胞均有阳性表达,但繁殖期牦牛ColⅣ在近管腔面生精细胞显示有强阳性表达,平均吸光度统计表明繁殖期显著高于繁殖间期(P<0.01);HSPG在不同繁殖期牦牛睾丸精原细胞及支持细胞强阳性表达,繁殖间期平均吸光度检测极显著低于繁殖期(P<0.01)。上述研究结果表明,高原环境中成年牦牛睾丸间质组织纤维成分随繁殖季节变化并不明显,睾丸组织LN合成与不同繁殖季节间质细胞分泌功能变化相关,繁殖期ColⅣ合成增强更有利于精子生成,HSPG的显著增加主要与精原细胞及支持细胞分化相关。

This study was conducted to characterize the histochemical characteristics and distribution of extracellular matrix proteins：laminin（LN）,type Ⅳ collagen（Col Ⅳ） and heparan sulfate proteoglycans（HSPG） in adult yak testis during different breeding seasons by Masson trichromatic dyeing,Gomori silver impregnation and immunohistochemical SP technique combined with Image-Pro Plus（IPP）statistical methods.The observations of light microscope found that the interstitial tissues were abundant with collagen and reticular fiber both in the breeding season and non-breeding season adult yak testis.Immunostaining analysis appeared that the LN was present,being similar in Sertoli cells and spermatogenic cells in both different seasons,and the relative expression of LN was almost the same between two different seasons although the expression in leydig cells during breeding season was reduced obviously（P〈0.05）.The immunoreactivity for Col Ⅳwere seen in Sertolicells and spermatogenic cells in both seasons but the Col Ⅳdistribution near the lumen of seminiferous tubule in breeding yak showed to be strong positive,and the relative expression of Col Ⅳ in the breeding season yak testis tissue was more higher than that in non-breeding seasons（P〈0.01）.In the adult yak testis, the HSPG expression was mainly strong in Sertoli cells and spermatogonium,and the average abso r bance of HSPG expression was decreased significantly in non-breeding season（P〈0.01）.Taken together, the changes of testis connective tissues in different breeding seasons are not obvious in yaks which live in plateau environment.The synthesis of LN changes with the secretion of leydig cells of different breeding season.In breeding season,the higher distribution of Col Ⅳ are closely related to a bettercondition of spermatogenesis and the expression of HSPG is significant in the sertoli cell and sperma togonia differentiation.