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EXPRESSION AND LOCALIZATION OF SMAD4 PROTEIN IN RAT TESTIS DURING THE POSTNATAL DEVELOPMENT

机译:发育后大鼠睾丸中SMAD4蛋白的表达和定位

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摘要

Objective. To study expression and localization of Smad4 protein, the common- mediator Smad, which is one of intracellular signaling molecules of transforming growth factor-β (TGF-β) family, in rat testis during postnatal development. Methods. In this study, whole testes were collected from S.D rats aged 3 days, 7 days, 14 days and 28 days, and adult respectively. We examined the cellular localization and developmental change of Smad4 in rat testis by immunohistochemical ABC method with glucose oxidase- DAB- nickel enhancement technique; the quantitative analysis of the immunostaining by the image analysis system; the Smads proteins coexistence in the adult rat testis by the double immune staining for CD14- Smad4 and Smad2 -Smad4; the protein expression of Smad during rat testicular development by means of Western blots. Results. The protein of Smad4 was present in rats from 3 days of age to adulthood, and the im-munolocalization 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. The result of image and statistical analysis showed that generally, there was a tendency that the expression of Smad4 in the testes increased gradually with the rats developing maturation. Conclusion. Our data provide direct evidence for the molecular mechanism of TGF-β action in rat testes during postnatal development and spermatogenesis of rats.
机译:目的。为了研究Smad4蛋白的表达和定位,其是产后发育期大鼠睾丸中Smad的共同媒介,Smad是转化生长因子β(TGF-β)家族的细胞内信号分子之一。方法。在这项研究中,从年龄分别为3天,7天,14天和28天的成年SD大鼠中收集整个睾丸。采用葡萄糖氧化酶-DAB-镍增强技术的免疫组织化学ABC法检测了大鼠睾丸中Smad4的细胞定位和发育变化。通过图像分析系统对免疫染色进行定量分析;通过CD14-Smad4和Smad2-Smad4的双重免疫染色,在成年大鼠睾丸中Smads蛋白共存; Western blot检测大鼠睾丸发育过程中Smad蛋白的表达结果。 Smad4的蛋白存在于3天大到成年的大鼠中,并且免疫定位在任何时间点仅位于间质组织中具有负核的Leydig细胞的细胞质中。在生殖细胞中未检测到表达。图像和统计分析结果表明,一般而言,随着大鼠的成熟,Smad4在睾丸中的表达有逐渐增加的趋势。结论。我们的数据提供了大鼠出生后发育和精子发生过程中TGF-β作用于大鼠睾丸的分子机制的直接证据。

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