title: Effects of naloxone benzoylhydrazone on native and recombinant nociceptin/orphanin FQ receptors.
abstract: We have studied the effects of naloxone benzoylhydrazone (NalBzoH) at recombinant human OP4 receptors expressed in Chinese hamster ovary (CHO) cells (CHOhOP4) and native OP4 sites in isolated tissues from various species. In CHOhOP4 membranes, nociceptin (NC) and NalBzoH displaced [125I]Tyr14-NC with pKi values of 10.1 and 7.3. In the presence of 100 microM GDP, NC stimulated GTPgamma35S binding (pEC50 = 8.5). NalBzoH was ineffective but antagonized the effects of NC (pA2 = 6.9). At 5 microM GDP, there was an increase in potency (pEC50 = 9.3) and efficacy (4.3-fold) of NC. NalBzOH was a partial agonist (pEC50 = 7.0, Emax = 13% relative to NC). In CHOhOP4 cells, NC and NalBzoH inhibited cAMP formation with pEC50 and Emax values of 9.8 and 100% and 6.0 and 44%, respectively. In the rat vas deferens, NalBzoH (10 microM) did not modify electrically induced twitches but competitively antagonized the inhibitory action of NC (pA2 = 6.2). In the mouse vas deferens (mVD) and guinea pig ileum (gpI), NalBzoH inhibited twitches with pEC50 and Emax values of 7.6 and 78% and 8.5 and 77%, respectively. The effect of 3 microM NalBzoH was fully inhibited by 3 microM naloxone in mVD and 30 microM in gpI. Under these conditions, NalBzoH antagonized the actions of NC in both preparations with pA2 values of 6.3 and 6.8, respectively. Collectively, these data demonstrate that NalBzoH is a nonselective OP4 ligand with system-dependent behaviour.
authors.affiliation: Department of Experimental and Clinical Medicine, Neuroscience Centre, University of Ferrara, Italy.
authors.forename: R
authors.initials: R
authors.lastname: Bigoni
authors.forename: G
authors.initials: G
authors.lastname: Caò
authors.forename: A
authors.initials: A
authors.lastname: Rizzi
authors.forename: H
authors.initials: H
authors.lastname: Okawa
authors.forename: D
authors.initials: D
authors.lastname: Regoli
authors.forename: D
authors.initials: D
authors.lastname: Smart
authors.forename: D G
authors.initials: DG
authors.lastname: Lambert
mesh_terms: D000818:Animals; D016466:CHO Cells / Q000187:drug effects / Q000378:metabolism; D006224:Cricetinae; D004305:Dose-Response Relationship, Drug; D004558:Electric Stimulation; D006168:Guinea Pigs; D006801:Humans; D066298:In Vitro Techniques; D008297:Male; D051379:Mice; D009270:Naloxone / Q000031:analogs & derivatives* / Q000378:metabolism / Q000494:pharmacology*; D009292:Narcotic Antagonists; D051381:Rats; D017207:Rats, Sprague-Dawley; D011957:Receptors, Opioid / Q000819:agonists / Q000378:metabolism; D011994:Recombinant Proteins / Q000819:agonists / Q000037:antagonists & inhibitors* / Q000378:metabolism; D000094942:Nociceptin Receptor
chemical_list: D009292:Narcotic Antagonists; D011957:Receptors, Opioid; D011994:Recombinant Proteins; C058380:naloxone benzoylhydrazone; D009270:Naloxone; D000094942:Nociceptin Receptor