Endocrinology, Vol 136, 1828-1834, Copyright © 1995 by Endocrine Society

ARTICLES

Signaling properties of mouse and human corticotropin-releasing factor (CRF) receptors: decreased coupling efficiency of human type II CRF receptor

Y Xiong, LY Xie and AB Abou-Samra
Endocrine Unit, Massachusetts General Hospital, Boston 02114, USA.

CRF is the primary neuroregulator of the function of the hypothalamic- pituitary-adrenal axis. We have recently cloned a mouse CRF receptor (mCRF-R) complementary DNA (cDNA) from an AtT-20 cell cDNA library by polymerase chain reaction. To compare the functions of mCRF-R to those of the human type I and type II CRF receptors (hCRF-RI and hCRF-RII), cDNAs were cloned into the expression vector pcDNA1 and transfected into COS-7 cells. CRF binding and CRF-stimulated cAMP accumulation as well as phosphoinositide hydrolysis were measured. Scatchard analysis of the binding of 125I-labeled [Tyr0]r/hCRF ([125I]CRF) to COS-7 cells expressing mCRF-R and hCRF-RI cDNAs revealed the same apparent Kd (9 nM). In contrast, the apparent binding Kd for hCRF-RII was 20 nM CRF. Maximal stimulatory concentrations (1 microM) of rat/human CRF-(1-41) (r/hCRF) increased cAMP accumulation in COS-7 cells transfected with mCRF-R, hCRF-RI, and hCRF-RII cDNA plasmid (10 micrograms each) from basal values of 8-19 pmol/10(5) cells.15 min to 84 +/- 10, 87 +/- 16, and 45 +/- 16 pmol/10(5) cells.15 min, respectively. The EC50 values of r/hCRF-stimulated cAMP accumulation in COS-7 cells expressing mCRF-R and hCRF-RI cDNAs were similar at 0.4 +/- 0.2 and 0.7 +/- 0.2 nM, respectively. Conversely, the EC50 of r/hCRF-stimulated cAMP accumulation in hCRF-RII-transfected COS-7 cells was 47.5 +/- 18.9 nM. As the level of expression of hCRF-RII was lower than that of hCRF-RI, we compared r/hCRF-stimulated cAMP accumulation in COS-7 cells expressing low and high levels of hCRF-RI. The EC50 for r/hCRF- stimulated cAMP accumulation in COS-7 cells transfected with hCRF-RI did not change when receptor expression was varied by a factor of 1- to 8.4-fold. In contrast, the EC50 for r/hCRF-stimulated cAMP accumulation mediated by hCRF-RII was at least 100-fold higher than that mediated by the hCRF-RI in COS-7 cells, which suggests poor coupling between hCRF- RII and adenylate cyclase. Inositol phosphate (IP) levels were also determined in mCRF-R, hCRF-RI, and hCRF-RII cDNA-transfected COS-7 cells stimulated with increasing concentrations of r/hCRF. r/hCRF- stimulated IPs accumulation was dose dependent in COS-7 cells expressing mCRF-R and hCRF-RI using 100 and 1000 nM r/hCRF. Concentrations of 10 (or less) nM r/hCRF had no effect on IP generation. hCRF-RII did not mediate stimulation of IP even at 1000 nM r/hCRF.(ABSTRACT TRUNCATED AT 400 WORDS)

This article has been cited by other articles:

				D. Markovic, N. Papadopoulou, T. Teli, H. Randeva, M. A. Levine, E. W. Hillhouse, and D. K. Grammatopoulos Differential Responses of Corticotropin-Releasing Hormone Receptor Type 1 Variants to Protein Kinase C Phosphorylation J. Pharmacol. Exp. Ther., December 1, 2006; 319(3): 1032 - 1042. [Abstract] [Full Text] [PDF]

				E. W. Hillhouse and D. K. Grammatopoulos The Molecular Mechanisms Underlying the Regulation of the Biological Activity of Corticotropin-Releasing Hormone Receptors: Implications for Physiology and Pathophysiology Endocr. Rev., May 1, 2006; 27(3): 260 - 286. [Abstract] [Full Text] [PDF]

				J. Cao, C. L. Cetrulo, and T. C. Theoharides Corticotropin-Releasing Hormone Induces Vascular Endothelial Growth Factor Release from Human Mast Cells via the cAMP/Protein Kinase A/p38 Mitogen-Activated Protein Kinase Pathway Mol. Pharmacol., March 1, 2006; 69(3): 998 - 1006. [Abstract] [Full Text] [PDF]

				C. Benou, Y. Wang, J. Imitola, L. VanVlerken, C. Chandras, K. P. Karalis, and S. J. Khoury Corticotropin-Releasing Hormone Contributes to the Peripheral Inflammatory Response in Experimental Autoimmune Encephalomyelitis J. Immunol., May 1, 2005; 174(9): 5407 - 5413. [Abstract] [Full Text] [PDF]

				P. J. Peeters, H. W. Gohlmann, I. Van den Wyngaert, S. M. Swagemakers, L. Bijnens, S. U. Kass, and T. Steckler Transcriptional Response to Corticotropin-Releasing Factor in AtT-20 Cells Mol. Pharmacol., November 1, 2004; 66(5): 1083 - 1092. [Abstract] [Full Text] [PDF]

				R. T. Hinkle, E. Donnelly, D. B. Cody, S. Samuelsson, J. S. Lange, M. B. Bauer, M. Tarnopolsky, R. J. Sheldon, S. C. Coste, E. Tobar, et al. Activation of the CRF 2 receptor modulates skeletal muscle mass under physiological and pathological conditions Am J Physiol Endocrinol Metab, October 1, 2003; 285(4): E889 - E898. [Abstract] [Full Text] [PDF]

				I. Assil-Kishawi and A. B. Abou-Samra Sauvagine Cross-links to the Second Extracellular Loop of the Corticotropin-releasing Factor Type 1 Receptor J. Biol. Chem., August 30, 2002; 277(36): 32558 - 32561. [Abstract] [Full Text] [PDF]

				I. Q. Assil and A. B. Abou-Samra N-glycosylation of CRF receptor type 1 is important for its ligand-specific interaction Am J Physiol Endocrinol Metab, November 1, 2001; 281(5): E1015 - E1021. [Abstract] [Full Text] [PDF]

				S. R. Smith, L. de Jonge, M. Pellymounter, T. Nguyen, R. Harris, D. York, S. Redmann, J. Rood, and G. A. Bray Peripheral Administration of Human Corticotropin-Releasing Hormone: A Novel Method to Increase Energy Expenditure and Fat Oxidation in Man J. Clin. Endocrinol. Metab., May 1, 2001; 86(5): 1991 - 1998. [Abstract] [Full Text]

				E. Karteris, D. Grammatopoulos, H. Randeva, and E. W. Hillhouse Signal Transduction Characteristics of the Corticotropin-Releasing Hormone Receptors in the Feto-Placental Unit J. Clin. Endocrinol. Metab., May 1, 2000; 85(5): 1989 - 1996. [Abstract] [Full Text]

				P. L. M. Dahia and A. B. Grossman The Molecular Pathogenesis of Corticotroph Tumors Endocr. Rev., April 1, 1999; 20(2): 136 - 155. [Abstract] [Full Text]

				C. J. Rossant, R. D. Pinnock, J. Hughes, M. D. Hall, and S. McNulty Corticotropin-Releasing Factor Type 1 and Type 2{alpha} Receptors Regulate Phosphorylation of Calcium/Cyclic Adenosine 3',5'-Monophosphate Response Element-Binding Protein and Activation of p42/p44 Mitogen-Activated Protein Kinase Endocrinology, April 1, 1999; 140(4): 1525 - 1536. [Abstract] [Full Text]

				L. Y. Xie and A. B. Abou-Samra Epitope Tag Mapping of the Extracellular and Cytoplasmic Domains of the Rat Parathyroid Hormone (PTH)/PTH-Related Peptide Receptor Endocrinology, November 1, 1998; 139(11): 4563 - 4567. [Abstract] [Full Text] [PDF]

				N. Heinrich, M. R. Meyer, J. Furkert, A. Sasse, M. Beyermann, W. Bonigk, and H. Berger Corticotropin-Releasing Factor (CRF) Agonists Stimulate Testosterone Production in Mouse Leydig Cells through CRF Receptor-1 Endocrinology, February 1, 1998; 139(2): 651 - 658. [Abstract] [Full Text] [PDF]

				L. Gnessi, A. Fabbri, and G. Spera Gonadal Peptides as Mediators of Development and Functional Control of the Testis: An Integrated System with Hormones and Local Environment Endocr. Rev., August 1, 1997; 18(4): 541 - 609. [Abstract] [Full Text] [PDF]

				Y. Aoki, Y. Iwasaki, M. Katahira, Y. Oiso, and H. Saito Regulation of the Rat Proopiomelanocortin Gene Expression in AtT-20 Cells. I: Effects of the Common Secretagogues Endocrinology, May 1, 1997; 138(5): 1923 - 1929. [Abstract] [Full Text] [PDF]

				V. Wu, M. Yang, J. A. McRoberts, J. Ren, R. Seensalu, N. Zeng, M. Dagrag, M. Birnbaumer, and J. H. Walsh First Intracellular Loop of the Human Cholecystokinin-A Receptor Is Essential for Cyclic AMP Signaling in Transfected HEK-293 Cells J. Biol. Chem., April 4, 1997; 272(14): 9037 - 9042. [Abstract] [Full Text] [PDF]

				C. H. Tsai-Morris, E. Buczko, Y. Geng, A. Gamboa-Pinto, and M. L. Dufau The Genomic Structure of the Rat Corticotropin Releasing Factor Receptor. A MEMBER OF THE CLASS II G PROTEIN-COUPLED RECEPTORS J. Biol. Chem., June 14, 1996; 271(24): 14519 - 14525. [Abstract] [Full Text] [PDF]