SOX5蛋白对公鸡精子发生和精子活力的作用及其定位

摘要

本研究旨在探明SOX5蛋白在家禽睾丸发育中的表达规律,分析其在睾丸发育和精子活力调控中的潜在作用.采用RT-qPCR和Western blot技术检测0、5、15、40和60周龄北京油鸡公鸡睾丸中SOX5 mRNA和蛋白的表达,并比较其在40周龄正常公鸡与弱精子症公鸡睾丸中的表达差异,通过免疫组化技术对SOX5蛋白在睾丸组织中的表达进行定位.结果表明,SOX5 mRNA和蛋白在不同周龄公鸡的睾丸中均有表达,且差异显著(P＜0.05).SOX5 mRNA和蛋白表达量趋势一致;随着周龄的增加,其表达呈现先上升后降低的趋势:0和5周龄SOX5蛋白主要在精原细胞和支持细胞中表达;15周龄SOX5蛋白在支持细胞、各级精母细胞、圆形精子细胞和成熟精子中均高表达;40周龄表达量有所降低.SOX5基因和蛋白的表达趋势与公鸡性成熟和繁殖机能衰退的趋势一致,即性成熟前后和繁殖高峰期表达量较高,在性发育前和生精机能减退时表达量较低.此外,40周龄弱精子症公鸡睾丸中,SOX5的表达显著低于正常公鸡(P＜0.05).综上表明,SOX5的表达量与公鸡睾丸发育水平和精子活力存在一定的相关,可能对其具有重要调控作用.%The aim of the present study was to investigate the expression of SOX5 in the testis of chickens and identify its possible roles in testis development and sperm motility regulation.The mRNA and protein expression of SOX5 in the testis of roosters at 0,5,15,40 and 60 weeks of age were determined by RT-qPCR and Western blot,respectively.Its cellular localization in the testis was determined by immunohistochemistry.The expression of SOX5 in the testis of normal and asthenospermia roosters at 40 weeks of age was also compared.The results showed that SOX5 mRNA and protein were differentially expressed in the testis at different ages (P＜0.05).The expression pattern of SOX5 protein was in accordance with the mRNA.The immunohistochemical staining revealed that SOX5 was expressed in the Sertoli cells and spermatogonia at 0 and 5 weeks of age.SOX5 was high-expressed in Sertoli cells,spermatocytes,secondary spermatocytes,round and elongated spermatids,and mature sperm in the testis at 15 weeks of age.The expression started to decrease at 40 weeks.The expression pattern of SOX5 was in accordance with the sexual development and recession of the roosters,i.e.high-expressed around the sexual maturation and reproduction peak and low-expressed before the puberty and sexual recession.In addition,the expression of SOX5 in the asthenospermia roosters was lower than the normal ones at 40 weeks of age (P＜0.05).In conclusion,this study indicated that the expression of SOX5 was associated with testis development and sperm motility in chickens,suggesting its regulatory roles in these processes.