摘要
探讨转化生长因子-β超家族肽类的细胞内信号转导分子Smad4蛋白在发育不同阶段大鼠睾丸的表达与分布。分别选用出生后3、7、14、28天以及成年大鼠,应用免疫组织化学ABC法结合葡萄糖氧化酶-DAB-硫酸镍铵增强技术及蛋白质免疫印迹技术,检测Smad4蛋白在大鼠睾丸的表达、定位和发育变化,并通过图像分析技术对免疫组织化学结果进行统计学分析。结果显示,发育各个阶段的间质细胞中都有较强的表达,免疫阳性产物位于细胞质内,而各级生精细胞则无阳性反应,且随着大鼠睾丸发育阶段的变化而蛋白表达量逐渐增多,为TGF-β超家族成员在精子发生和发育过程中的分子机理提供了直接证据。
Ill order to shed light on the mechanisms of TGF-β action in the testis, we examined the expression and function of Smad4 protein, the common-mediator Smads, which is one of intracellular signaling molecules of TGF-β superfamily members, in rat testis during postnatal development. Whole testes were collected from SD rats aged 3 days, 7 days, 14 days and 28 days, and adult. In this study, we examined, by means of western blots, the protein expression of Smad4 during rat testicular development and its cellular localization by immunohistochemical ABC method with glucose oxidase-DAB-nickel enhancement technique. The results showed that the protein of Smad4 was present in rats from 3 days of age to adulthood, and the im-munoreactivity for Smad4 was exclusively localized to the cytoplasm of Leydig cells with negative nuclei in the interstitial tissue at any time point. No expression was detected in germ cells. Therefore, our data provide evidence for the molecular mechanism of TGF-β action in rat testes during postnatal development and spermatoge-nesis of rats.
出处
《实验生物学报》
CSCD
北大核心
2002年第1期7-14,共8页
Acta Biologiae Experimentalis Sinica
基金
国家自然科学基金(No.39870109)
全军医药卫生科研基金(No.98M106
01MA183)