National Demonstration Center for Experimental Fisheries Science Education, Shanghai Ocean University, Shanghai 201306, China.
Key Laboratory of Exploration and Utilization of Aquatic Genetic Resources, Ministry of Education, Shanghai Ocean University, Shanghai 201306, China.
Int J Mol Sci. 2019 Apr 8;20(7):1736. doi: 10.3390/ijms20071736.
Luteinizing hormone (LH), a pituitary gonadotropin, coupled with LH receptor (LHR) is essential for the regulation of the gonadal maturation in vertebrates. Although LH homolog has been detected by immunocytochemical analysis, and its possible role in ovarian maturation was revealed in decapod crustacean, so far there is no molecular evidence for the existence of LHR. In this study, we cloned a novel homolog (named ) from the Chinese mitten crab . The complete sequence of the cDNA was 2775bp, encoding a protein of 924 amino acids, sharing 71% amino acids identity with the ant LHR. expression was found to be high in the ovary, while low in testis, gill, brain, and heart, and no expression in the thoracic ganglion, eye stalk, muscle, and hepatopancreas. Quantitative PCR revealed that the expression level of mRNA was significantly higher in the ovaries in previtellogenic (Pvt), late vitellogenic (Lvt), and germinal vesicle breakdown (GVBD) stages than that in the vitellogenic (Mvt) and early vitellogenic (Evt) stages ( < 0.05), and, the highest and the lowest expression were in Lvt, and Evt, respectively. The strong signal was mainly localized in the ooplasm of Pvt oocyte as detected by in situ hybridization. The crab GnRH homolog can significantly induce the expression of mRNA at 36 hours post injection in vivo ( < 0.01), suggesting that may be involved in regulating ovarian development through GnRH signaling pathway in the mitten crab.
促黄体生成素（LH）是一种垂体促性腺激素，与 LH 受体（LHR）结合对于脊椎动物性腺成熟的调节至关重要。尽管已经通过免疫细胞化学分析检测到 LH 同源物，并且在十足目甲壳动物中揭示了其在卵巢成熟中的可能作用，但迄今为止，尚无 LHR 存在的分子证据。在这项研究中，我们从中华绒螯蟹中克隆了一种新的 同源物（命名为 ）。 的 cDNA 全长为 2775bp，编码 924 个氨基酸的蛋白质，与蚂蚁的 LHR 具有 71%的氨基酸同一性。 在卵巢中表达量较高，而在精巢、鳃、脑和心脏中表达量较低，在胸神经节、眼柄、肌肉和肝胰腺中无表达。定量 PCR 显示，在卵黄发生前期（Pvt）、晚期卵黄发生期（Lvt）和核膜破裂期（GVBD）卵巢中 mRNA 的表达水平明显高于卵黄发生期（Mvt）和早期卵黄发生期（Evt）（ < 0.05），并且 Lvt 和 Evt 期的表达水平最高和最低。原位杂交检测结果表明，在 Pvt 卵母细胞的卵质中观察到强烈的信号。在体内注射 GnRH 同源物 36 小时后，蟹 GnRH 同源物可以显著诱导 mRNA 的表达（ < 0.01），这表明 在中华绒螯蟹中可能通过 GnRH 信号通路参与调节卵巢发育。
