Kakar S S, Rahe C H, Neill J D
Department of Physiology and Biophysics, University of Alabama at Birmingham 35294.
Domest Anim Endocrinol. 1993 Oct;10(4):335-42. doi: 10.1016/0739-7240(93)90037-c.
Receptors for gonadotropin releasing hormone (GnRH), located in the cell membranes of adeno-hypophysial gonadotropes, mediate the action of GnRH to stimulate the secretion of the gonadotropic hormones (LH and FSH). In the present studies, we have isolated a GnRH receptor cDNA from bovine pituitary, determined its primary structure, and studied the regulation of its gene expression. The cDNA is composed of 1326 nucleotides and encodes a protein containing 328 amino acids. The GnRH receptor of cattle, like that in humans and mice, is a seven transmembrane receptor and has structural characteristics homologous with the family of G protein-coupled receptors. It exhibits 91% identity at the amino acid level with the human and 86% identity with mouse and rat receptors. Northern blot analysis of the RNA from bovine pituitary, probed with 32P-labeled bovine GnRH receptor cDNA, revealed the presence of four different transcripts (5.0, 3.5, 2.5 and 1.5 kb) in the pituitary of which the 5.0 kb form was most abundant. Using the reverse transcriptase/PCR technique, we detected expression of GnRH receptor mRNA in the pituitary but not in any other extrapituitary tissues such as the hypothalamus, hippocampus, testis, corpus luteum, ovary (containing follicles), myoendometrium, adrenal, kidney, liver and spleen. Higher levels of GnRH receptor mRNA were found in the pituitaries of steers than in cohort bulls, suggesting regulation of GnRH receptor gene expression by testicular steroids.
促性腺激素释放激素(GnRH)受体位于腺垂体促性腺细胞的细胞膜上,介导GnRH刺激促性腺激素(LH和FSH)分泌的作用。在本研究中,我们从牛垂体中分离出GnRH受体cDNA,确定了其一级结构,并研究了其基因表达的调控。该cDNA由1326个核苷酸组成,编码一种含328个氨基酸的蛋白质。牛的GnRH受体与人类和小鼠的一样,是一种七跨膜受体,具有与G蛋白偶联受体家族同源的结构特征。它在氨基酸水平上与人类GnRH受体有91%的同一性,与小鼠和大鼠的受体有86%的同一性。用32P标记的牛GnRH受体cDNA探测牛垂体RNA的Northern印迹分析显示,垂体中存在四种不同的转录本(5.0、3.5、2.5和1.5 kb),其中5.0 kb的形式最为丰富。使用逆转录酶/PCR技术,我们在垂体中检测到了GnRH受体mRNA的表达,但在下丘脑、海马体、睾丸、黄体、卵巢(含卵泡)、肌内膜、肾上腺、肾脏、肝脏和脾脏等任何其他垂体外组织中均未检测到。发现阉牛垂体中的GnRH受体mRNA水平高于同龄公牛,提示睾丸类固醇对GnRH受体基因表达有调控作用。