人睾丸特异性基因蛋白抗体制备及其在无精症睾丸组织中的表达

Preparation of the antibody against human TSC21 protein and its expression characteristics in testis from patients with azoospermia

目的探讨人睾丸特异性基因(TSC21)蛋白在无精子症患者睾丸组织中的表达特征。方法构建人pET32a-TSC21表达载体,转化到感受态BL21细菌,诱导表达并纯化TSC21蛋白;免疫小鼠,制备抗人的TSC21多克隆抗体;应用免疫组化方法检测TSC21在健康者睾丸组织和生精阻滞患者睾丸组织中的表达差异。结果成功制备人TSC21的鼠源性多克隆抗体;TSC21蛋白在健康者睾丸组织中主要定位于精母细胞和圆形精子细胞;在生精阻滞的无精症患者中表达减弱。结论 TSC21蛋白对于睾丸发育和精子发生可能具有重要作用,其表达降低可能是男性不育症发生的重要原因之一。

Objective To investigate the expression characteristics of TSC21 in testis from patients with azoospermia.Methods Human TSC21 gene was constructed recombinant expression plasmid pET32a vectors hTSC21,and transferred into BL21.IPTG induced the gene expressed.The protein of interest was purified,then we immunized the mice for preparing antiserum,and detected the expression and localization of TSC21 proteins and the difference of expression in testis from fertile men and the patients with azoospermia by immunohistochemistry.Results Anti TSC21 polyclonal antibody was generated successfully;Immunohistochemistry showed that TSC21 was predominantly located in spermatocytes and round spermatids.In the testis from patients with azoospermia,the expression of the protein was significantly decreased in spermatocytes and round spermatids.Conclusion TSC21 may participate in testicular development and spermatogenesis,and its abnormal expression may be one of the important factors in male infertility.