Department of Zoology, University of Otago, 340 Great King Street, Dunedin 9016, New Zealand.
J Steroid Biochem Mol Biol. 2013 Nov;138:464-70. doi: 10.1016/j.jsbmb.2013.02.003. Epub 2013 Feb 10.
Steroidogenic acute regulatory protein (StAR) mRNA levels in the eel ovary were assayed by quantitative PCR and related to plasma steroid levels throughout oogenesis in order to shed light on the previously considered 'aberrant' prematurational increase in plasma levels of estradiol-17β (E2). Total ovarian StAR transcript abundance mirrored circulating levels of E2, but not of 11-ketotestosterone (11KT). The study was complemented by evaluation of in vitro effects of follicle-stimulating hormone (FSH) on ovarian StAR transcript abundance and on short-term ('acute') radiolabelled pregnenolone-supported steroid metabolism by ovarian fragments to understand how the production of steroids during previtellogenic oocyte growth is regulated. We observed a significant effect of FSH on StAR mRNA levels within 24h of incubation, but these were no longer evident by 4 days of culture. Unexpectedly, FSH had no effect on substrate-supported steroidogenesis, as comparable yields of steroid products were detected using semi-quantitative HPLC and scintillation counting. We conclude that the eel ovarian follicle can respond to FSH from a very early stage of development (early oil droplet stage) by increasing StAR mRNA levels, but that there is no evidence for acute effects of FSH on bioactive steroid production downstream of cytochrome P450 side-chain cleavage. Furthermore, the prematurational increase in StAR mRNA in vivo is in keeping with general teleost models and is likely to be a 'normal' response to reaching advanced stages of development.
通过定量 PCR 测定了鳗鱼卵巢中类固醇生成急性调节蛋白 (StAR) mRNA 的水平,并将其与整个卵发生过程中的血浆类固醇水平相关联,以便阐明先前被认为是“异常的”血浆雌二醇-17β(E2)水平提前升高的现象。总卵巢 StAR 转录丰度与循环 E2 水平相关,但与 11-酮睾酮(11KT)无关。该研究还通过评估卵泡刺激素(FSH)对卵巢 StAR 转录丰度的体外影响以及对卵巢片段短期(“急性”)放射性标记孕烯醇酮支持的类固醇代谢的影响来补充,以了解卵母细胞生长过程中类固醇的产生是如何调节的。我们观察到 FSH 在孵育 24 小时内对 StAR mRNA 水平有显著影响,但在培养 4 天后这种影响不再明显。出乎意料的是,FSH 对底物支持的类固醇生成没有影响,因为使用半定量 HPLC 和闪烁计数检测到可比量的类固醇产物。我们得出的结论是,鳗鱼卵巢滤泡可以在发育的早期(早期油滴阶段)对 FSH 做出反应,增加 StAR mRNA 水平,但没有证据表明 FSH 对细胞色素 P450 侧链裂解下游的生物活性类固醇产生有急性影响。此外,体内 StAR mRNA 的提前增加与一般的硬骨鱼模型一致,可能是对达到发育后期的“正常”反应。