EphA2基因在藏绵羊睾丸和附睾中的表达与定位  

Expression and localization of EphA2 gene in testis and epididymis of Tibetan sheep

作　　者：雒瑞瑞 李讨讨 苏亮宁 LUO Ruirui;LI Taotao;SU Liangning(Institute of Pasture and Green Agriculture,Gansu Academy of Agricultural Sciences,Lanzhou 730070,Gansu,China;College of Animal Science and Technology,Gansu Agricultural University,Lanzhou 730070,Gansu,China)

机构地区：[1]甘肃省农业科学院畜草与绿色农业研究所,甘肃兰州730070 [2]甘肃农业大学动物科学技术学院,甘肃兰州730070

出　　处：《草业科学》2025年第3期762-771,共10页Pratacultural Science

基　　金：甘肃省农业科学院农业科技创新专项计划资助项目(2022GAAS60);甘肃省科技计划资助项目(23JRRA1338)。

摘　　要：睾丸的发育和精子生成受到多种因子复杂而精细的调控。促红细胞生成素肝细胞受体A2(EphA2)是一种参与细胞间信号传导的跨膜蛋白。为了解析EphA2基因在藏绵羊精子发生和成熟中的表达特征及发挥的生物学作用,选取3月龄、1岁龄和3岁龄的健康雄性藏绵羊各4只,通过实时荧光定量聚合酶链式反应(PCR)、蛋白质印迹法和免疫荧光染色分别检测了EphA2基因在不同发育阶段睾丸和附睾的表达和细胞分布特征。mRNA检测结果显示,随着年龄的增加,EphA2基因在睾丸、附睾头、体和尾中的表达均呈下降趋势。结果显示,随着年龄的增加EphA2蛋白在藏绵羊睾丸和附睾头中的表达水平显著升高(P<0.01),而在附睾体、尾中显著减少(P<0.01)(3月龄和1岁龄附睾体除外)。EphA2蛋白主要分布于生殖母细胞(3月龄)、精原细胞(3岁组)和精细胞(1岁和3岁组);在附睾中,其主要存在于3月龄组附睾头和体中的疏松结缔组织,1岁龄组和3岁龄组的附睾头、附睾体及整个发育阶段附睾尾的假复层柱状纤毛上皮细胞。综上表明,EphA2可能在生精细胞的早期发育、精子细胞的分化以及精子成熟中起重要调控作用,这为深入解析EphA2基因在藏绵羊乃至其他绵羊精子形成和成熟中的分子机制提供了参考。Testicular development and sperm production are regulated by several complex and precise factors.Erythropoietin-producing hepatocellular A2(EphA2)is a transmembrane protein involved in intercellular signaling.Four healthy male Tibetan sheep aged 3 months,1 year,and 3 years were selected to investigate the expression characteristics and biological functions of the EphA2 gene in developmental Tibetan sheep testis and epididymis.The expression and cellular distribution characteristics of the EphA2 gene in testis and epididymis at different developmental stages were detected by quantitative real-time Polymerase Chain Reaction(PCR),western blot,and immunofluorescence(IF)staining,respectively.The results of mRNA detection showed that the expression of the EphA2 gene in the testis,epididymis head,body,and tail tended to decrease with age.Results showed that with increasing age,the expression of EphA2 protein significantly increased in the testis and epididymal head(P<0.01)while significantly decreasing in the epididymal body and tail(P<0.01)(except for the epididymis at 3 months and 1 year).EphA2 protein was mainly localized in the gonocytes of the 3-month-old testes,spermatogonia of the 3-year-old group,and spermatocytes of the 1-and 3-year-old groups.In the epididymis,it is mainly present in the loose connective tissue of the epididymal head and body of the 3-month-old group and the pseudostratified columnar ciliated epithelial cells of the epididymal head and body of the 1-and 3-year-old groups,as well as in the entire developmental stage of the epididymal tail.In conclusion,EphA2 may play an important regulatory role in early germ cell development,sperm differentiation,and sperm maturation.This provides a clue for further understanding of the molecular mechanisms of the EphA2 gene in spermatogenesis and maturation in Tibetan and other sheep.

关 键 词：藏绵羊 睾丸 附睾 表达 定位 睾丸发育 

分 类 号：R73[医药卫生—肿瘤]