Monkey corticotropin-releasing factor1 receptor: Complementary DNA cloning and pharmacological characterization.
The full-length complementary DNA (cDNA) of monkey corticotropin-releasing factor type 1 (CRF1) receptor was isolated from a rhesus monkey (Macaca mulatta) amygdala cDNA library. The cloned monkey CRF1 receptor cDNA has 2,374 bp with an open reading frame encoding a 415-amino acid protein. The sequence of the monkey CRF1 receptor cDNA showed a high degree of sequence identity with other species of CRF1 receptors, and being 99.5% identical to human CRF1 receptors. When monkey CRF1 was expressed into COS-7 cells, high specific binding of [125I]-ovine CRF was observed. CRF and CRF-related peptides inhibited [125I]-ovine CRF binding in a concentration-dependent manner. IC50 values of ovine CRF, human/rat CRF, sauvagine and urotensin I were 23.5 +/- 7.4, 22.7 +/- 10.8, 27.5 +/- 12.3 and 14.2 +/- 7.0 nM, respectively. CRF1 receptor specific antagonists, such as CP-154,526, SC241 and CRA1000, also inhibited the [125I]-ovine CRF binding, with IC50 values of 3.9 +/- 0.4, 43.5 +/- 8.0 and 19.8 +/- 2.0 nM, respectively. GTP and its nonhydrolyzed analogue, GTPgammaS, reduced [125I]-ovine CRF binding, while ATP had a negligible effect, thereby indicating that the monkey CRF1 receptor belongs to a family of G-protein coupled receptors. CRF and its related peptides increased cyclic AMP formation concentration-dependently in COS-7 cells transiently expressing the monkey CRF1 receptor. Monkey CRF1 was expressed abundantly in the pituitary, cerebral cortex, hippocampus, amygdala and cerebellum. Thus the monkey CRF1 receptor and the human CRF1 receptor have similar molecular and pharmacological characteristics.
Oshida
Yuichi
Y
Psychiatric Diseases and Pain Research, Medicinal Pharmacology Laboratory, Medicinal Research Laboratories, Taisho Pharmaceutical Co., Ltd., 1-403 Yoshino-cho, Kita, Saitama, Saitama 331-9530, Japan. y.oshida@po.rd.taisho.co.jp
Ikeda
Yoko
Y
Chaki
Shigeyuki
S
Okuyama
Shigeru
S
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D019556:COS Cells; D002522:Chlorocebus aethiops; D003346:Corticotropin-Releasing Hormone / Q000031:analogs & derivatives / Q000378:metabolism*; D017931:DNA Primers; D018076:DNA, Complementary / Q000235:genetics; D004587:Electrophoresis, Agar Gel; D016244:Guanosine 5'-O-(3-Thiotriphosphate) / Q000494:pharmacology; D006160:Guanosine Triphosphate / Q000494:pharmacology; D007457:Iodine Radioisotopes; D008253:Macaca mulatta / Q000235:genetics*; D008969:Molecular Sequence Data; D011485:Protein Binding / Q000187:drug effects; D018019:Receptors, Corticotropin-Releasing Hormone / Q000235:genetics* / Q000378:metabolism; D017422:Sequence Analysis, DNA; D017385:Sequence Homology; D014162:Transfection; D000099052:CRF Receptor, Type 1
D017931:DNA Primers; D018076:DNA, Complementary; D007457:Iodine Radioisotopes; D018019:Receptors, Corticotropin-Releasing Hormone; D016244:Guanosine 5'-O-(3-Thiotriphosphate); D000099052:CRF Receptor, Type 1; D006160:Guanosine Triphosphate; D003346:Corticotropin-Releasing Hormone