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通过微阵列分析确定的新生小鼠睾丸中的雄激素调节转录本。

Androgen-regulated transcripts in the neonatal mouse testis as determined through microarray analysis.

作者信息

Zhou Qing, Shima James E, Nie Rong, Friel Patrick J, Griswold Michael D

机构信息

Center for Reproductive Biology, School of Molecular Biosciences, Washington State University, Pullman, Washington 99164, USA.

出版信息

Biol Reprod. 2005 Apr;72(4):1010-9. doi: 10.1095/biolreprod.104.035915. Epub 2004 Dec 15.

Abstract

Androgens are required for normal spermatogenesis in mammalian testes. These hormones directly regulate testicular somatic cells that, in turn, support germ cell differentiation. However, the identity of genes under androgen regulation in the testis are not well known. In the present study, neonatal male mice (8 days postpartum) treated by testosterone propionate (TP) were used to study androgen action in the testis as evidenced by alterations in gene expression. Mice were treated with 0.5 mg of TP or dihydrotestosterone (DHT) or vehicle (oil), and testes were harvested 4, 8, and 16 h after treatment. Global gene expression was monitored by microarray analysis. Real-time reverse transcription-polymerase chain reaction was performed to confirm the microarray results. The methodology was verified by confirming the presence of previously characterized TP-regulated genes, including Pem in Sertoli cells and Cyp17a1 in Leydig cells. No significant differences in gene expression were found between TP- and DHT-treated samples. Microarray analysis identified 141, 119, and 109 up-regulated genes at 4, 8 and 16 h after TP treatment, respectively, and 83, 99, and 111 down-regulated genes at the same corresponding time points. The androgen regulation of the selected gene was verified further using testes from flutamide-treated adult mice and isolated Sertoli cells in culture. The data generated in the present study may serve as a foundation for hypothesis-driven research and provide insights regarding gene networks and pathways under androgen control in the testis.

摘要

雄激素是哺乳动物睾丸正常精子发生所必需的。这些激素直接调节睾丸体细胞,而睾丸体细胞又支持生殖细胞的分化。然而,睾丸中受雄激素调节的基因的身份尚不清楚。在本研究中,使用丙酸睾酮(TP)处理的新生雄性小鼠(产后8天)来研究睾丸中的雄激素作用,基因表达的变化证明了这一点。小鼠分别用0.5 mg的TP或双氢睾酮(DHT)或载体(油)处理,并在处理后4、8和16小时采集睾丸。通过微阵列分析监测全局基因表达。进行实时逆转录-聚合酶链反应以确认微阵列结果。通过确认先前已鉴定的TP调节基因的存在来验证该方法,包括支持细胞中的Pem和睾丸间质细胞中的Cyp17a1。在TP处理和DHT处理的样品之间未发现基因表达的显著差异。微阵列分析分别在TP处理后4、8和16小时鉴定出141、119和109个上调基因,以及在相同相应时间点分别有83、99和111个下调基因。使用氟他胺处理的成年小鼠的睾丸和培养的分离支持细胞进一步验证了所选基因的雄激素调节作用。本研究中产生的数据可为假设驱动的研究奠定基础,并提供有关睾丸中雄激素控制下的基因网络和途径的见解。

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