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克隆、药理学特征分析和大西洋鲑(Salmo salar L.)核孕激素受体的表达分析。

Cloning, pharmacological characterization, and expression analysis of Atlantic salmon (Salmo salar L.) nuclear progesterone receptor.

机构信息

Department of Biology, Faculty of Science, Utrecht University, Padualaan 8, 3584 CH Utrecht, The Netherlands.

出版信息

Reproduction. 2011 Apr;141(4):491-500. doi: 10.1530/REP-10-0224. Epub 2011 Jan 18.

DOI:10.1530/REP-10-0224
PMID:21245280
Abstract

To better understand the role(s) of progestogens during early stages of spermatogenesis, we carried out studies on the nuclear progesterone receptor (Pgr) of the Atlantic salmon. Its open-reading frame shows the highest similarity with other piscine Pgr proteins. When expressed in mammalian cells, salmon Pgr exhibited progestogen-specific, dose-dependent induction of reporter gene expression, with 17α,20β-dihydroxy-4-pregnen-3-one (DHP) showing the highest potency. We then analyzed testicular pgr mRNA and DHP plasma levels in animals during the onset of spermatogenesis, which were exposed to natural light or to constant light, to induce significant differences in testis growth. Grouping of the animals according to their progress through spermatogenesis showed that testicular pgr mRNA levels as well as DHP plasma levels first increased when germ cells had reached the stage of late type B spermatogonia and further increased when entered meiosis, i.e. when spermatocytes were present. However, in situ hybridization studies revealed that pgr mRNA expression was restricted to Sertoli cells, with a strong signal in Sertoli cells contacting type A/early type B spermatogonia, while Sertoli cells contacting larger germ cell clones with further differentiated stages (e.g. late type B spermatogonia) were less intensely/not stained. We conclude that the increase in pgr mRNA levels per pair of testis reflects, at least in part, the increased number of Sertoli cells enveloping type A and early type B spermatogonia. We propose that Sertoli cell-expressed Pgr may mediate DHP-stimulated early steps in spermatogenesis in Atlantic salmon, such as an increase in the number of new spermatogonial cysts.

摘要

为了更好地理解孕激素在精子发生早期阶段的作用,我们对大西洋三文鱼的核孕激素受体(Pgr)进行了研究。其开放阅读框与其他鱼类 Pgr 蛋白具有最高的相似性。当在哺乳动物细胞中表达时,三文鱼 Pgr 表现出孕激素特异性、剂量依赖性的报告基因表达诱导,17α,20β-二羟-4-孕烯-3-酮(DHP)显示出最高的效力。然后,我们分析了在诱导睾丸生长显著差异的情况下,处于精子发生起始阶段的动物的睾丸 pgr mRNA 和 DHP 血浆水平,这些动物暴露于自然光或持续光照下。根据它们通过精子发生的进展对动物进行分组表明,当生殖细胞达到晚期 B 型精原细胞阶段时,睾丸 pgr mRNA 水平以及 DHP 血浆水平首先增加,当进入减数分裂时,即当精母细胞存在时,进一步增加。然而,原位杂交研究表明,pgr mRNA 表达仅限于支持细胞,与 A 型/早期 B 型精原细胞接触的支持细胞具有强烈信号,而与具有进一步分化阶段(例如晚期 B 型精原细胞)的更大生殖细胞克隆接触的支持细胞信号较弱/未染色。我们得出结论,每对睾丸中 pgr mRNA 水平的增加至少部分反映了包裹 A 型和早期 B 型精原细胞的支持细胞数量的增加。我们提出,支持细胞表达的 Pgr 可能介导了 DHP 刺激的大西洋三文鱼精子发生的早期步骤,例如新精原细胞囊泡数量的增加。

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