HSPA2和SPATA6在彭波半细毛羊睾丸组织中的细胞定位及表达差异分析

Expression and localization of HSPA2 and SPATA6 in testis of Pengbo semi-fine wool sheep

热休克蛋白A2(HSPA2)是热休克蛋白HSP70家族重要成员,能够促进睾丸支持细胞增殖;精子发生相关基因6(SPATA6)能够调控肌球蛋白合成的微丝运输系统,影响精子细胞骨架形成。在雄性哺乳动物生殖过程中,两者均起重要作用。为研究HSPA2和SPATA6在雄性彭波半细毛羊生殖过程中的表达规律和细胞定位,以不同年龄阶段(3月龄、12月龄和3岁龄)彭波半细毛羊睾丸组织为研究对象,运用实时荧光定量PCR(real time quantitative PCR,qRT-PCR)、Western blot和免疫组织化学染色法(IHC)从转录和翻译水平检测了HSPA2和SPATA6在不同年龄阶段彭波半细毛羊睾丸组织的表达水平和分布模式,分析预测HSPA2和SPATA6在彭波半细毛羊睾丸发育和精子发生过程中的调控作用。结果表明:12月龄和3岁龄彭波半细毛羊睾丸组织中HSPA2 mRNA的相对表达量以及蛋白表达量极显著高于3月龄(P<0.01);12月龄SPATA6 mRNA的相对表达量以及蛋白表达量极显著高于3月龄(P<0.01),显著高于3岁龄(P<0.05)。不同年龄阶段彭波半细毛羊睾丸组织的支持细胞和间质细胞中HSPA2和SPATA6蛋白均有分布。初步推断,HSPA2和SPATA6的表达水平变化在调控彭波半细毛羊精子发生过程和促进精子成熟中具有重要作用。

Heat shock protein A2(HSPA2)is an important member of the HSP70 family,promoting the proliferation of testicular support cells.Spermatogene-related gene 6(SPATA6)can regulate the microfilament transport system of myosin synthesis and affect the formation of the sperm cytoskeleton,both of which play important roles in the reproductive process of male mammals.This study investigated the regulation mechanisms of heat shock protein A2(HSPA2)and spermatogenesis-related gene 6(SPATA6)in the reproductive process of male animals to provide a scientific basis for the breeding of Pengbo semi-fine wool sheep.Pengbo semi-fine wool sheep of different ages(3 months,12 months,and 3 years)were taken as the research object.Real-time quantitative PCR(qRT-PCR),western blot,and immunohistochemical staining were used to detect the expression and distribution patterns of HSPA2 and SPATA6 in the testicular tissues of Pengbo semi-fine wool sheep of different ages from transcription and translation levels and to analyze the biological functions of HSPA2 and SPATA6 in the testicular development and spermatogenesis of Pengbo semi-fine wool sheep.Results showed that the relative expression of HSPA2 mRNA and protein in the testis of Pengbo semi-fine wool sheep at the age of 12 months and 3 years was significantly higher than that at the age of 3 months(P<0.01);moreover,they tended to be stable after sexual maturity.The relative expression of SPATA6 mRNA and protein at 12 months old was significantly higher than that at 3 months old(P<0.01)and significantly higher than that at 3 years old(P<0.05).HSPA2 and SPATA6 proteins were distributed in sertoli cells and interstitial cells of the testicular tissue of Pengbo semi-fine wool sheep at different ages.We speculate that HSPA2 and SPATA6 play an important role in regulating spermatogenesis and promoting sperm maturation.