title: Characterisation using microphysiometry of CRF receptor pharmacology.
abstract: We have assessed the utility of the Cytosensor microphysiometer for studying the pharmacology of recombinant CRF receptors. Chinese hamster ovary cells stably expressing the human CRF1 or CRF2 receptor were perfused in the Cytosensor with bicarbonate-free Hams F12 (pH 7.4) containing 0.2% bovine serum albumin. The rank order of potencies of agonist peptides were CRF = sauvagine = urocortin = urotensin at CRF1 (pEC50 values 11.16 +/- 0.17, 11.37 +/- 0.14, 11.43 +/- 0.09 and 11.46 +/- 0.13; n = 4), and urocortin = sauvagine > urotensin > CRF at CRF2 (pEC50 values 10.88 +/- 0.12, 10.44 +/- 0.05, 9.36 +/- 0.12 and 8.53 +/- 0.07; n = 7-9). alpha-Helical CRF (9-41) was a competitive antagonist at the CRF2 receptor (pK(B) = 6.99 +/- 0.08, n = 4), but was a partial agonist at the CRF1 receptor (pEC50 = 6.85 +/- 0.08, Emax = 33%, n = 3). CP 154,526 was a competitive antagonist at the CRF1 receptor (pK(B) = 8.17 +/- 0.05, n = 6), but was inactive at the CRF2 receptor. These data are consistent with established CRF receptor pharmacology and show that the Cytosensor is a viable method for assessing the functional activity of CRF-receptor agonists and antagonists.
authors.affiliation: Parke-Davis Neuroscience Research Centre, Cambridge University Forvie Site, UK. darren_2_smart@sbphrd.com
authors.forename: D
authors.initials: D
authors.lastname: Smart
authors.forename: A
authors.initials: A
authors.lastname: Coppell
authors.forename: C
authors.initials: C
authors.lastname: Rossant
authors.forename: M
authors.initials: M
authors.lastname: Hall
authors.forename: A T
authors.initials: AT
authors.lastname: McKnight
mesh_terms: D000136:Acid-Base Equilibrium; D029845:Amphibian Proteins; D000818:Animals; D016466:CHO Cells; D002417:Cattle; D003346:Corticotropin-Releasing Hormone / Q000493:pharmacokinetics* / Q000494:pharmacology; D006224:Cricetinae; D006801:Humans; D008564:Membrane Potentials; D010446:Peptide Fragments / Q000493:pharmacokinetics*; D036361:Peptide Hormones; D010455:Peptides / Q000494:pharmacology; D051381:Rats; D018019:Receptors, Corticotropin-Releasing Hormone / Q000819:agonists / Q000037:antagonists & inhibitors / Q000378:metabolism*; D054832:Urocortins; D014579:Urotensins / Q000494:pharmacology; D014665:Vasodilator Agents / Q000494:pharmacology; D000099052:CRF Receptor, Type 1
chemical_list: 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; D014665:Vasodilator Agents; D000099052:CRF Receptor, Type 1; C025727:sauvagine; D003346:Corticotropin-Releasing Hormone; C029676:urotensin I; C045089:corticotropin releasing hormone (9-41)