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克隆两种促性腺激素受体并研究其在斜带石斑鱼卵子发生过程中的不同表达谱。

Molecular cloning of two gonadotropin receptors and their distinct mRNA expression profiles in daily oogenesis of the wrasse Pseudolabrus sieboldi.

机构信息

Laboratory of Marine Biology, Faculty of Agriculture, Kyushu University, Higashi-Ku, Fukuoka, Japan.

出版信息

Gen Comp Endocrinol. 2011 Jun 1;172(2):268-76. doi: 10.1016/j.ygcen.2011.03.012. Epub 2011 Mar 21.

Abstract

In fish, asynchronous development of ovarian follicles, the simultaneous advance of vitellogenesis and oocyte maturation in one ovary, is a rational reproductive strategy to spawn consecutively in one spawning season. In this study, to clarify the mode of action of follicle stimulating hormone (FSH) and luteinizing hormone (LH) in asynchronous ovarian follicle development in daily egg production, we cloned cDNAs of the follicle-stimulating hormone receptor (FSHR) and luteinizing hormone receptor (LHR) in the bambooleaf wrasse (Pseudolabrus sieboldi), which exhibits clear diurnal spawning rhythms over 1 month. In addition, different developmental stages of ovarian follicles were isolated from whole ovaries at various daily time points on 1 day in the spawning season, and mRNA expression levels of FSHR and LHR were analyzed. Sequence analysis showed distinct differences in the number of putative leucine-rich repeats at the extracellular domain between FSHR and LHR, suggesting a difference in ligand-specificity. Real-time PCR analyses revealed that FSHR mRNA was highly expressed in early yolk-stage follicles but decreased at the end of vitellogenesis. In contrast, the expression of LHR mRNA was maintained at low levels in vitellogenic stage follicles but markedly elevated at the end of the vitellogenic and early migratory nucleus stages, thereafter markedly dropping in the late migratory nucleus stage. The present results suggest that co-regulation of vitellogenesis and oocyte maturation in one ovary is controlled by the stage-distinctive expression levels of FSHR and LHR mRNA in ovarian follicles, and daily switching of sensitivity from FSH to LH is required for daily egg production.

摘要

在鱼类中,卵巢滤泡的异步发育,即一个卵巢中卵黄发生和卵母细胞成熟的同时进行,是一种在一个繁殖季节中连续产卵的合理生殖策略。在这项研究中,为了阐明促卵泡激素(FSH)和促黄体生成素(LH)在每日产卵中异步卵巢滤泡发育中的作用模式,我们克隆了在竹荚鱼(Pseudolabrus sieboldi)中滤泡刺激激素受体(FSHR)和黄体生成素受体(LHR)的 cDNA,竹荚鱼在一个月的时间内表现出明显的昼夜产卵节律。此外,我们从繁殖季节的 1 天的不同日常时间点从整个卵巢中分离出不同发育阶段的卵巢滤泡,并分析了 FSHR 和 LHR 的 mRNA 表达水平。序列分析表明,FSHR 和 LHR 的细胞外结构域中的假定亮氨酸丰富重复数存在明显差异,提示配体特异性的差异。实时 PCR 分析显示,FSHR mRNA 在早期卵黄发生阶段高度表达,但在卵黄发生结束时降低。相比之下,LHR mRNA 在卵黄发生阶段的滤泡中保持低水平表达,但在卵黄发生和早期迁移核阶段结束时显著升高,此后在晚期迁移核阶段显著下降。这些结果表明,一个卵巢中卵黄发生和卵母细胞成熟的共同调控是由卵巢滤泡中 FSHR 和 LHR mRNA 的阶段特异性表达水平控制的,并且需要从 FSH 到 LH 的每日敏感性转换来实现每日产卵。

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