Gupta C
Department of Pediatrics (Endocrinology), Children's Hospital of Pittsburgh, Rangos Research Center, University of Pittsburgh, Pennsylvania 15213.
Biol Reprod. 1994 Aug;51(2):283-9. doi: 10.1095/biolreprod51.2.283.
Differentiation of the mammalian reproductive tract into the male phenotype is dependent on the presence of functional testes secreting testosterone during in utero development. Recently, we identified a protein fraction (72 kDa) from the developing reproductive tract of the fetal male mouse that is able to mimic the effect of testosterone in stabilizing the Wolffian duct in vitro, suggesting a role played by this protein in mediating the effect of testosterone during Wolffian duct differentiation. In the present study, we further evaluated the masculinizing role of this protein by determining the specific appearance of the 72-kDa protein in relation to reproductive tract differentiation. Using Western blot and immunohistochemical analysis, we demonstrate that the protein was expressed specifically in the male reproductive tract and not in male non-reproductive organs, namely, lung, liver, or kidney. Within the male reproductive tract, the Wolffian duct derivatives demonstrated a high level of expression of the protein, whereas the urogenital sinus produced only a low level of reaction to this antibody in Western analysis and no reaction in the histochemical analysis. There was a gradual rise in expression of the protein in the male reproductive tract during the critical period of Wolffian duct differentiation. The female reproductive tract produced only a trace immune reaction in Western analysis and no reaction in immunohistochemical analysis. Thus, the results support the hypothesis that the 72-kDa protein plays a role in Wolffian duct differentiation.
哺乳动物生殖道向雄性表型的分化取决于子宫内发育期间分泌睾酮的功能性睾丸的存在。最近,我们从雄性胎鼠发育中的生殖道中鉴定出一种蛋白质组分(72 kDa),它能够在体外模拟睾酮在稳定中肾管方面的作用,这表明该蛋白质在中肾管分化过程中介导睾酮的作用。在本研究中,我们通过确定72 kDa蛋白质与生殖道分化相关的特定出现情况,进一步评估了该蛋白质的雄性化作用。使用蛋白质印迹和免疫组织化学分析,我们证明该蛋白质在雄性生殖道中特异性表达,而在雄性非生殖器官,即肺、肝或肾中不表达。在雄性生殖道内,中肾管衍生物显示出该蛋白质的高水平表达,而泌尿生殖窦在蛋白质印迹分析中对该抗体仅产生低水平反应,在组织化学分析中无反应。在中肾管分化的关键时期,雄性生殖道中该蛋白质的表达逐渐增加。雌性生殖道在蛋白质印迹分析中仅产生微量免疫反应,在免疫组织化学分析中无反应。因此,结果支持了72 kDa蛋白质在中肾管分化中起作用的假设。