Molecular cloning, characterization, and gene expression behavior of glucocorticoid and mineralocorticoid receptors from the Chinese alligator (Alligator sinensis)
Molecular cloning, characterization, and gene expression behavior of glucocorticoid and mineralocorticoid receptors from the Chinese alligator (Alligator sinensis)
复制标题
扬子鳄糖皮质激素和盐皮质激素受体的分子克隆、表征和基因表达行为
DOI：
10.1002/jez.b.23015
复制
发表时间：
2020-12-11
影响因子：
2.2
通讯作者：
Wu，Xiaobing
中科院分区：
文献类型：
--
作者：
Izaz，Ali;Pan，Tao;Wu，Xiaobing
扬子鳄是中国特有的鳄鱼物种。本研究采用套式聚合酶链式反应(PCR)和快速扩增基因末端扩增(RACE)技术，从扬子鳄肾脏中分离得到糖皮质激素和盐皮质激素受体基因。糖皮质激素受体(GR)基因有2,343个碱基对，编码780个氨基酸，而盐皮质激素受体(MR)基因全长约2958 ，编码985个氨基酸。用实时荧光定量聚合酶链式反应检测信使RNA水平的分布。在卵巢和肾脏中表达最高，提示这些受体可能参与了扬子鳄的基本细胞功能或应激反应。此外，还利用RT-qPCR技术分析了扬子鳄早期胚胎发育过程中肾脏、肝脏和心脏中GR和MR基因转录产物的丰度。早期肾脏、肝脏和心脏中GR和MR的mRNA水平表明，它们可能参与了早期胚胎的转录调控，并激活了胎儿组织中许多精确的发育效应。我们还检测了GR和MR蛋白在肝脏胚胎发育阶段的表达，发现GR和MR蛋白既限于细胞核，也限于细胞质。不同胚胎发育阶段肝脏蛋白质表达水平存在极显著差异。综上所述，我们的结果显示了GR和MR的完整编码区、它们的特征以及这两个基因在胚胎发育过程中的mRNA和蛋白表达。中华绒毛。本研究为进一步研究GR和MR INA的不同功能提供了必要的信息。中华绒毛。
The Chinese alligator is an endemic crocodilian species in China. We isolated and obtained the glucocorticoid and mineralocorticoid receptor genes coding from the kidney ofAlligator sinensisby nested polymerase chain reaction (PCR) and rapid amplification of cDNA ends (RACE). The glucocorticoid receptor (GR) gene has 2343 base pairs encoding 780 amino acids, while the mineralocorticoid receptor (MR) gene is 2958 bp in length encoding 985 amino acids. Quantitative real‐time PCR was used to detect the distribution of messenger RNA (mRNA) levels. The maximum mRNA expressions were observed in the ovary and kidney, suggesting that these receptors may be involved in basic cellular functions or stress response of alligators. Besides this, RT‐qPCR was performed to analyze the abundance of GR and MR mRNA transcripts in early embryonic development of the Chinese alligator in the kidney, liver, and heart. The mRNA levels of GR and MR at earlier stages in kidney, liver, and heart indicates that they might involve in the transcriptional regulation of early embryos and activate many precise developmental effects in fetal tissues. We also measured the protein expression in the liver embryonic developmental stages and found that the GR and MR proteins were restricted to both the nuclei and cytoplasm. The protein expression levels in the liver at different embryonic developmental stages have extremely prominent differences. Taken together, our results showed the full coding regions of GR and MR, their characteristics, and embryonic developmental mRNA and protein expressions of both genes inA. sinensis. This study could provide the necessary information for further investigating the diverse functions of GR and MR inA. sinensis.