Molecular cloning and functional expression of the mouse CRF2(a) receptor splice variant.
The mouse corticotropin-releasing factor (CRF) type 2a receptor (CRF2(a)) splice variant was cloned by a PCR-based approach. The corresponding cDNA was found to encode a 411-amino acid polypeptide with highest sequence homology to the rat CRF2(a) receptor. By semiquantitative reverse transcriptase PCR (RT-PCR) analysis, the CRF2(b) mRNA was mainly found in the heart and skeletal muscle with only low level expression in the brain. In contrast, CRF2(a) mRNA was restricted to the brain with major expression sites in the cortex, hippocampus, hypothalamus and telencephalon. Binding and cyclic AMP stimulation studies showed a similar ligand selective profile for both mCRF2 receptor splice variants. A notable exception however, was urotensin I which displayed a approximately 3-fold higher affinity for the CRF2(a) receptor and also stimulated cyclic AMP production in mCRF2(a)-transfected cells with a approximately 3-fold higher potency than in mCRF2(b)-transfected cells. These data show that the mouse like other mammalian species expresses two ligand-selective CRF2 receptor splice variants and that the mCRF2(a) receptor is the predominant central CRF2 receptor in the mouse.
Dautzenberg
Frank M
FM
F. Hoffmann-La Roche AG, Pharma Research, 4070 Basel, Switzerland. fdautzen@prdbe.jnj.com
Higelin
Jacqueline
J
Wille
Sandra
S
Brauns
Olaf
O
D017398:Alternative Splicing / Q000235:genetics*; D000595:Amino Acid Sequence; D029845:Amphibian Proteins; D000818:Animals; D001921:Brain / Q000378:metabolism; D002460:Cell Line; D003001:Cloning, Molecular; D003346:Corticotropin-Releasing Hormone / Q000494:pharmacology; D000242:Cyclic AMP / Q000096:biosynthesis / Q000378:metabolism; D020869:Gene Expression Profiling; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000494:pharmacology; D036361:Peptide Hormones; D010455:Peptides / Q000494:pharmacology; D011485:Protein Binding; D018019:Receptors, Corticotropin-Releasing Hormone / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D016415:Sequence Alignment; D014162:Transfection; D054832:Urocortins; D014579:Urotensins / Q000494:pharmacology
D029845:Amphibian Proteins; C101531:CRF receptor type 2; D010446:Peptide Fragments; D036361:Peptide Hormones; D010455:Peptides; D018019:Receptors, Corticotropin-Releasing Hormone; D054832:Urocortins; D014579:Urotensins; C104416:astressin; C025727:sauvagine; D003346:Corticotropin-Releasing Hormone; C029676:urotensin I; D000242:Cyclic AMP