Ritsumeikan Global Innovation Research Organization, Ritsumeikan University, Kusatsu, Shiga 525-8577, Japan.
Gen Comp Endocrinol. 2013 Sep 15;191:65-73. doi: 10.1016/j.ygcen.2013.06.002. Epub 2013 Jun 13.
Fish Sertoli cells play a critical role in spermatogenesis by mediating androgen and progestogen signaling. Their hormonal response, however, considerably differ among species. Therefore it would be ideal to use Sertoli cells originated from the fish of interest to investigate the effects of hormones as well as endocrine disrupting chemicals (EDCs). The aim of this study was to investigate the responses to reproductive hormones and EDCs of a Sertoli cell line that we established from an endemic cyprinid Gnathopogon caerulescens. As the Sertoli cell line expressed endogenous androgen and progestogen receptors, we were able to detect hormone responses by transfecting only a reporter vector (pGL4.36) expressing luciferase under the control of the mouse mammary tumor virus-long terminal repeat (MMTV-LTR) promoter into the cell line. Unlike previous reporter gene assays using fish steroid hormone receptors expressed in mammalian cell lines, luciferase activities were induced by the fish specific androgen (11-ketotestosterone) and progestogen (17α,20β-dihydroxy-4-pregnen-3-one), but not by testosterone and progesterone, at physiologically relevant concentrations. Furthermore, we found 4-nonylphenol (NP) but not bisphenol A showed strong anti-androgenic effects, implying that NP may have direct anti-androgenic effects on fish Sertoli cells in vivo. This is the first evidence, to the best of our knowledge, of anti-androgenic effects of NP in a fish Sertoli cell line. In addition, neither NP nor BPA showed anti-progestogenic effects. These results suggest that the Sertoli cell line established from the fish of interest can be a useful in vitro tool for investigating the mechanisms of reproductive hormones and EDCs in the specific fish.
鱼类支持细胞在精子发生中发挥关键作用,通过介导雄激素和孕激素信号。然而,它们的激素反应在不同物种之间有很大的差异。因此,使用感兴趣的鱼类来源的支持细胞来研究激素以及内分泌干扰化学物质(EDCs)的影响是理想的。本研究的目的是研究我们从一种特有鲤科鱼类 Gnathopogon caerulescens 中建立的支持细胞系对生殖激素和 EDC 的反应。由于支持细胞系表达内源性雄激素和孕激素受体,我们能够通过仅转染表达荧光素酶的报告基因载体(pGL4.36)来检测激素反应,该载体受小鼠乳腺肿瘤病毒长末端重复(MMTV-LTR)启动子的控制。与以前使用在哺乳动物细胞系中表达的鱼类类固醇激素受体的报告基因检测不同,在生理相关浓度下,只有鱼特异性雄激素(11-酮睾酮)和孕激素(17α,20β-二羟基-4-孕烯-3-酮)诱导了荧光素酶活性,而不是睾酮和孕酮。此外,我们发现 4-壬基酚(NP)而不是双酚 A 表现出强烈的抗雄激素作用,这表明 NP 可能对体内鱼类支持细胞具有直接的抗雄激素作用。这是我们所知的 NP 在鱼类支持细胞系中具有抗雄激素作用的第一个证据。此外,NP 或 BPA 均未显示出抗孕激素作用。这些结果表明,从感兴趣的鱼类中建立的支持细胞系可以成为研究生殖激素和 EDCs 在特定鱼类中的作用机制的有用体外工具。