title: Comparison of the effects of [Phe1psi(CH2-NH)Gly2]nociceptin(1-13)NH2 in rat brain, rat vas deferens and CHO cells expressing recombinant human nociceptin receptors.
abstract: Nociceptin(NC) is the endogenous ligand for the opioid receptor like-1 receptor (NC-receptor). [Phe1(psi)(CH2-NH)Gly2]Nociceptin(1-13)NH2 ([F/G]NC(1-13)NH2) has been reported to antagonize NC actions in peripheral guinea-pig and mouse tissues. In this study, we investigated the effects of a range of NC C-terminal truncated fragments and [F/G]NC(1-13)NH2 on NC receptor binding, glutamate release from rat cerebrocortical slices (rCX), inhibition of cyclic AMP accumulation in CHO cells expressing the NC receptor (CHO(NCR)) and electrically evoked contractions of the rat vas deferens (rVD). In radioligand binding assays, a range of ligands inhibited [125I]-Tyr14-NC binding in membranes from rCX and CHO(NCR) cells. As the peptide was truncated there was a general decline in pKi. [F/G]NC(1-13)NH2 was as potent as NC(1-13)NH2. The order of potency for NC fragments to inhibit cyclic AMP accumulation in whole CHO(NCR) cells was NCNH2> or =NC=NC(1-13)NH2>NC(1-12)NH2> >NC(1-11)NH2. [F/G]NC(1-13)NH2 was a full agonist with a pEC50 value of 8.65. NCNH2 and [F/G]NC(1-13)NH2 both inhibited K+ evoked glutamate release from rCX with pEC50 and maximum inhibition of 8.16, 48.5+/-4.9% and 7.39, 58.9+/-6.8% respectively. In rVD NC inhibited electrically evoked contractions with a pEC50 of 6.63. Although [F/G]NC(1-13)NH2, displayed a small (instrinsic activity alpha = 0.19) but consistent residual agonist activity, it acted as a competitive antagonist (pA2 6.76) in the rVD. The differences between [F/G]NC(1-13)NH2 action on central and peripheral NC signalling could be explained if [F/G]NC(1-13)NH2 was a partial agonist with high strength of coupling in the CNS and low in the periphery. An alternative explanation could be the existence of central and peripheral receptor isoforms.
authors.affiliation: University Department of Anaesthesia, Leicester Royal Infirmary.
authors.forename: H
authors.initials: H
authors.lastname: Okawa
authors.forename: B
authors.initials: B
authors.lastname: Nicol
authors.forename: R
authors.initials: R
authors.lastname: Bigoni
authors.forename: R A
authors.initials: RA
authors.lastname: Hirst
authors.forename: G
authors.initials: G
authors.lastname: Calo
authors.forename: R
authors.initials: R
authors.lastname: Guerrini
authors.forename: D J
authors.initials: DJ
authors.lastname: Rowbotham
authors.forename: D
authors.initials: D
authors.lastname: Smart
authors.forename: A T
authors.initials: AT
authors.lastname: McKnight
authors.forename: D G
authors.initials: DG
authors.lastname: Lambert
mesh_terms: D000818:Animals; D001667:Binding, Competitive; D016466:CHO Cells; D002540:Cerebral Cortex / Q000187:drug effects / Q000378:metabolism*; D006224:Cricetinae; D000242:Cyclic AMP / Q000378:metabolism; D004558:Electric Stimulation; D005260:Female; D018698:Glutamic Acid / Q000378:metabolism; D006801:Humans; D066298:In Vitro Techniques; D008024:Ligands; D008297:Male; D009131:Muscle, Smooth, Vascular / Q000187:drug effects / Q000378:metabolism / Q000502:physiology; D018847:Opioid Peptides / Q000494:pharmacology*; D010446:Peptide Fragments / Q000494:pharmacology*; D051381:Rats; D017208:Rats, Wistar; D011957:Receptors, Opioid / Q000096:biosynthesis*; D011994:Recombinant Proteins / Q000096:biosynthesis; D014649:Vas Deferens / Q000187:drug effects / Q000378:metabolism* / Q000502:physiology; D000094942:Nociceptin Receptor
chemical_list: D008024:Ligands; D018847:Opioid Peptides; D010446:Peptide Fragments; D011957:Receptors, Opioid; D011994:Recombinant Proteins; C114149:nociceptin (1-13)-NH2, Phe(1)-psi(CH2-NH)-Gly(2)-; D018698:Glutamic Acid; D000242:Cyclic AMP; D000094942:Nociceptin Receptor