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雌性锦龟(Chrysemys picta)肝脏中孕酮和雌激素受体的调节:与卵黄生成的关系。

Regulation of hepatic progesterone and estrogen receptors in the female turtle, Chrysemys picta: relationship to vitellogenesis.

作者信息

Custodia-Lora Noemí, Novillo Apolonia, Callard Ian P

机构信息

Department of Biology, Boston University, Boston, MA 02115, USA.

出版信息

Gen Comp Endocrinol. 2004 Apr;136(2):232-40. doi: 10.1016/j.ygcen.2003.12.016.

Abstract

Previous studies using the fresh water turtle Chrysemys picta have demonstrated the differential expression of the two progesterone receptor isoforms (PRA and PRB) in the liver during the turtle seasonal cycle, correlating with ER mRNA levels and hepatic vitellogenesis. During the inter-vitellogenic periods the ratio of PRB:PRA favors PRA, suggesting that the PRB:PRA ratio may be important in the regulation of vitellogenesis. Based on these and other studies we hypothesize that progesterone may have differential effects on the estrogen (E)-induced vitellogenin response (inhibitory or stimulatory), depending on the PRB:PRA ratio. In this study, we determined the expression pattern and the hormonal regulation of progesterone receptor (PR) isoforms in the liver, during the estrogen-induced vitellogenin response in female C. picta. Northern blot analysis was used to evaluate the changes in vitellogenin mRNA, estrogen receptor (ER) mRNA, and PR mRNA expression; Western blot to determine changes in PR isoform expression and a homologous ELISA for measurement of plasma vitellogenin. The expression of PR isoforms in the liver of female turtles at the mRNA and protein levels was differentially regulated by estradiol and progesterone. Estradiol treatment enhanced the transcription of PR mRNA isoforms and possibly translation and/or increased stability of PRB protein in the female turtle liver. In contrast, this hormone decreased PRA protein levels. Progesterone alone down-regulated progesterone receptor isoform A (PRA) and progesterone receptor isoform B (PRB) proteins equally, but did not affect PR mRNA levels. Estradiol markedly increased hepatic ER mRNA, vtg mRNA, and vtg in plasma, but this effect was not impacted significantly by progesterone. The results presented here indicate that in female turtles hepatic PRB:PRA ratios are markedly changed by estradiol treatment due primarily to a decrease in PRA. The change in the PRB:PRA ratio after hormonal treatment confirms that progesterone and estrogen exposure will be a determinant in the regulation of vitellogenesis, and, in turn, that the regulation of vitellogenesis will be determined by the ratio of PR isoforms and the physiological levels of steroid hormones.

摘要

此前使用淡水龟黄斑彩龟开展的研究表明,在龟的季节性周期中,肝脏中两种孕酮受体亚型(PRA和PRB)存在差异表达,这与雌激素受体mRNA水平及肝脏卵黄生成有关。在卵黄生成间期,PRB:PRA的比例有利于PRA,这表明PRB:PRA的比例可能在卵黄生成的调节中起重要作用。基于这些及其他研究,我们推测,孕酮可能根据PRB:PRA的比例,对雌激素(E)诱导的卵黄蛋白原反应产生不同影响(抑制或刺激)。在本研究中,我们确定了雌性黄斑彩龟在雌激素诱导的卵黄蛋白原反应过程中,肝脏中孕酮受体(PR)亚型的表达模式及激素调节情况。采用Northern印迹分析评估卵黄蛋白原mRNA、雌激素受体(ER)mRNA和PR mRNA表达的变化;采用Western印迹确定PR亚型表达的变化,并采用同源ELISA测定血浆卵黄蛋白原。雌二醇和孕酮对雌性龟肝脏中PR亚型在mRNA和蛋白质水平的表达进行了差异调节。雌二醇处理增强了PR mRNA亚型的转录,并可能促进了PRB蛋白的翻译和/或增加了其稳定性。相比之下,这种激素降低了PRA蛋白水平。单独使用孕酮可同等程度地下调孕酮受体亚型A(PRA)和孕酮受体亚型B(PRB)蛋白,但不影响PR mRNA水平。雌二醇显著增加了肝脏ER mRNA、卵黄蛋白原mRNA以及血浆中的卵黄蛋白原,但孕酮对这一效应没有显著影响。此处呈现的结果表明,在雌性龟中,主要由于PRA的减少,雌二醇处理显著改变了肝脏中PRB:PRA的比例。激素处理后PRB:PRA比例的变化证实,孕酮和雌激素暴露将是卵黄生成调节的一个决定因素,进而表明卵黄生成的调节将由PR亚型的比例和类固醇激素的生理水平决定。

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