Primary structure and functional expression of mouse pituitary and human brain corticotrophin releasing factor receptors
Primary structure and functional expression of mouse pituitary and human brain corticotrophin releasing factor receptors
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DOI：
10.1016/0014-5793(93)80427-v
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发表时间：
1993-11
期刊：
影响因子：
3.5
通讯作者：
N. Vita;P. Laurent;S. Lefort;P. Chalon;J. Lelias;M. Kaghad;G. Le Fur;D. Caput;P. Ferrara
中科院分区：
文献类型：
--
作者：
N. Vita;P. Laurent;S. Lefort;P. Chalon;J. Lelias;M. Kaghad;G. Le Fur;D. Caput;P. Ferrara
促肾上腺皮质激素释放因子（CRF）是控制垂体-肾上腺轴的主要下丘脑因子，但CRF及其受体的广泛垂体外分布表明这种神经肽在整合生物体对应激的整体生理和行为反应中发挥重要作用。我们通过在COS-7细胞中表达来自AtT 20小鼠垂体瘤细胞系的cDNA文库，克隆了CRF受体互补DNA（cDNA）。然后将克隆的小鼠cDNA用作探针，从人脑cDNA文库中分离人CRF受体cDNA。小鼠和人类cDNA均编码415个氨基酸的蛋白质，其具有97%的相同性，包含7个推定的G蛋白偶联受体特征性跨膜结构域。CRF受体与生长激素释放因子、血管活性肠肽、促胰液素、甲状旁腺激素和降钙素的受体具有同源性。转染小鼠CRF受体cDNA的COS-7细胞以高亲和力结合放射性标记的绵羊CRF，并通过细胞内cAMP的积累对CRF产生特异性反应。在AtT 20细胞和人皮质组织中检测到一个2.7kb的CRF受体mRNA。PCR分析还检测到人垂体、脑干和睾丸中的受体转录本。
Corticotrophin‐releasing factor (CRF) is the principal hypothalamic factor governing the pituitary‐adrenal axis, but the wide extra‐pituitary distribution of CRF and its receptors suggest a major role for this neuropeptide in the integration of the overall physiological and behavioral responses of an organism to stress. We have cloned a CRF receptor complementary DNA (cDNA) by expression in COS‐7 cells of a cDNA library from the AtT20 mouse pituitary tumour cell line. The cloned mouse cDNA was then used as a probe to isolate a human CRF receptor cDNA from a human brain cDNA library. The mouse and human cDNAs both encode 415 amino acid proteins that are 97% identical, containing seven putative transmembrane domains characteristic of G protein‐coupled receptors. The CRF receptor shows homology with the receptors for growth hormone‐releasing factor, vasoactive intestinal peptide, secretin, parathyroid hormone, and calcitonin. COS‐7 cells transfected with the mouse CRF receptor cDNA bind radiolabelled ovine CRF with high affinity and respond specifically to CRF by accumulation of intracellular cAMP. A 2.7 kb mRNA coding for the CRF receptor could be detected in AtT20 cells and human cortex tissue. PCR analysis also detected the receptor transcript in human pituitary, brainstem, and testis.