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窄头双髻鲨(Sphyrna tiburo)中的雄激素受体:cDNA克隆及在雄性生殖道中的组织特异性表达

Androgen receptors in the bonnethead, Sphyrna tiburo: cDNA cloning and tissue-specific expression in the male reproductive tract.

作者信息

Tyminski John P, Gelsleichter James J, Motta Philip J

机构信息

Center for Shark Research, Mote Marine Laboratory, 1600 Ken Thompson Parkway, Sarasota, FL 34236, USA.

Department of Biological Sciences, University of North Florida, 1 UNF Drive, Jacksonville, FL 32224, USA.

出版信息

Gen Comp Endocrinol. 2015 Dec 1;224:235-46. doi: 10.1016/j.ygcen.2015.08.018. Epub 2015 Aug 28.

Abstract

As demonstrated in past studies, androgens appear to play critical roles in regulating reproduction in male sharks. However, little is known about the cell-specific actions of androgens in these fishes. To address this, this study examined androgen targets in reproductive organs of a seasonally reproducing shark, the bonnethead (Sphyrna tiburo). A partial bonnethead AR cDNA clone was isolated and found to exhibit strong homology with known vertebrate ARs. Using RT-PCR and in situ hybridization, AR was found to be expressed in multiple cell types in the male bonnethead testis (premeiotic germ cells, Leydig-like interstitial cells, Sertoli cells, peritubular myoid cells, and mature spermatozoa) and gonadal ducts (stromal cells, luminal epithelial cells, mature spermatozoa). Furthermore, AR expression in these organs was found to vary temporally in relation to the seasonal reproductive cycle. Based on immunocytochemistry, the presence of AR protein in male bonnethead reproductive organs was largely consistent with patterns of AR gene expression with the single exception of mature spermatozoa, which exhibited consistently strong mRNA expression but only inconsistent and weak AR protein immunoreactivity. These results suggest important roles for androgens in regulating germ cell proliferation, hormone production, spermatid elongation, spermiation, and gonadal duct function in male bonnetheads. In addition, high abundance of AR mRNA in bonnethead spermatozoa suggest the potential for de novo protein synthesis following spermiation/copulation and/or a role for AR mRNA in early embryonic development, both of which have been proposed to explain the occurrence of mRNA transcripts in spermatozoa from various vertebrates.

摘要

正如过去的研究所表明的,雄激素似乎在调节雄性鲨鱼的生殖过程中发挥着关键作用。然而,关于雄激素在这些鱼类中的细胞特异性作用却知之甚少。为了解决这个问题,本研究检测了一种季节性繁殖鲨鱼——窄头双髻鲨(Sphyrna tiburo)生殖器官中的雄激素靶标。分离出了窄头双髻鲨AR cDNA的部分克隆,发现其与已知的脊椎动物AR具有高度同源性。使用逆转录聚合酶链反应(RT-PCR)和原位杂交技术,发现AR在雄性窄头双髻鲨睾丸的多种细胞类型(减数分裂前的生殖细胞、类睾丸间质细胞、支持细胞、睾丸肌样细胞和成熟精子)以及性腺管(基质细胞、管腔上皮细胞、成熟精子)中均有表达。此外,还发现这些器官中AR的表达随季节性生殖周期而发生时间变化。基于免疫细胞化学分析,雄性窄头双髻鲨生殖器官中AR蛋白的存在情况与AR基因表达模式基本一致,唯一的例外是成熟精子,其mRNA表达始终很强,但AR蛋白免疫反应性却不一致且较弱。这些结果表明,雄激素在调节雄性窄头双髻鲨的生殖细胞增殖、激素产生、精子细胞伸长、精子排放以及性腺管功能方面发挥着重要作用。此外,窄头双髻鲨精子中AR mRNA的高丰度表明,在精子排放/交配后可能进行从头蛋白质合成,和/或AR mRNA在早期胚胎发育中发挥作用,这两者都被认为可以解释各种脊椎动物精子中mRNA转录本的存在。

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