Wang L H, Zhang W, Ji J L, Gao Q X, Xiao S H, Wang F
Center of Embryo Engineering and Technology, College of Animal Science and Technology, Nanjing Agricultural University, Nanjing, Jiangsu, China.
Genet Mol Res. 2013 May 6;12(2):1490-500. doi: 10.4238/2013.May.6.2.
Liver receptor homologue 1 (Lrh-1) is a member of the nuclear receptor belonging to the second subfamily of the nuclear receptor family 5A (NR5A), also named NR5A2, which is important for lipid homeostasis, embryogenesis, and regulation of aromatics. The present study aimed to understand the sequence of ovine Lrh-1 and the expression traits in reproductive organ tissues. Initially, we cloned Lrh-1 from the liver of Hu sheep through degenerate primer of reverse transcription-polymerase chain reaction and rapid amplification of cDNA ends. Characteristic functional domains of DNA binding and ligand binding, conserved among transcription factors of the nuclear receptor superfamily, were identified in Lrh-1 of Hu sheep. The Lrh-1 protein levels in the tissues detected by Western blotting correlated significantly with the transcript levels measured by quantitative real-time polymerase chain reaction (qRT-PCR). To understand the Lrh-1 expression change in the hypothalamus and hypophysis during the estrous cycle, we analyzed the expression pattern of Lrh-1 mRNA and protein by qRT-PCR and Western blotting, respectively. This analysis revealed that Lrh-1 expression in the hypothalamus was highest during the metestrus phase, while the Lrh-1 level was similar during other phases. In the hypophysis, the expression was significantly different during the 4 phases of the estrous cycle but highest during the estrus phase, significantly correlating with FSH concentration. These results indicate that Lrh-1 expression is correlated with gonadotropic hormone secretion, influencing follicular formation in the ovary.
肝脏受体同源物1（Lrh-1）是核受体家族5A（NR5A）第二个亚家族的成员，也被称为NR5A2，对脂质稳态、胚胎发育和芳香族化合物的调节具有重要作用。本研究旨在了解绵羊Lrh-1的序列及其在生殖器官组织中的表达特征。首先，我们通过逆转录-聚合酶链反应的简并引物和cDNA末端快速扩增技术，从湖羊肝脏中克隆出Lrh-1。在湖羊的Lrh-1中鉴定出了核受体超家族转录因子中保守的DNA结合和配体结合特征性功能域。通过蛋白质印迹法检测的组织中Lrh-1蛋白水平与通过定量实时聚合酶链反应（qRT-PCR）测量的转录水平显著相关。为了了解发情周期中下丘脑和垂体中Lrh-1的表达变化，我们分别通过qRT-PCR和蛋白质印迹法分析了Lrh-1 mRNA和蛋白质的表达模式。该分析表明，下丘脑Lrh-1的表达在发情后期最高，而在其他阶段Lrh-1水平相似。在垂体中，发情周期的4个阶段表达存在显著差异，但在发情期最高，与促卵泡生成素浓度显著相关。这些结果表明，Lrh-1的表达与促性腺激素分泌相关，影响卵巢中的卵泡形成。
