title: Pertussis toxin, but not tyrosine kinase inhibitors, abolishes effects of U-50488H on [Ca2+]i in myocytes.
abstract: The effects of 10(-5) M trans-3,4-dichloro-N-[2-(1-pyrrolidinyl)cyclohexyl]benzeacetamidel (U-50488H), a kappa-opioid receptor agonist, on cytosolic Ca2+ concentration ([Ca2+]i) and the [Ca2+]i transient in quiescent and electrically stimulated rat ventricular myocytes, respectively, were determined after the cells had been pretreated with pertussis toxin (PTX) or a tyrosine kinase inhibitor (genistein or tyrphostin). The [Ca2+]i was determined with a spectrofluorometric method, with fura 2 as Ca2+ indicator. U-50488H at 10(-5) M itself induced a [Ca2+]i transient in the quiescent cells but inhibited the [Ca2+]i transient in electrically stimulated cells. The effects of 10(-5) M U-50488H on [Ca2+]i, which were blocked by a selective kappa-opioid receptor antagonist, nor-binaltorphimine (10(-6) M), were abolished after pretreatment with PTX (1 microg/ml) for 24 h, but not with genistein (10(-4) M) or tyrphostin (5 x 10(-5) M) for 30 min. 1-[6-[[(17b)-3-Methoxyestra-1,3,5(10)-trien-17-yl]amino]hexy l]-1H-pyrrole-2,5-dione (U-73122), an inhibitor of phospholipase C, at 10(-5) M, but not its inactive structural isomer 1-[6-[[(17b)-3-methoxyestra-1,3,5(10)-trien-17-yl]amino]hexy l]-2,5-pyrrolidinedione (U-73343), also blocked the Ca2+ responses to U-50488H. The results indicate that activation of phospholipase C on kappa-opioid receptor stimulation is via PTX-sensitive G proteins but does not involve protein tyrosine phosphorylation.
authors.affiliation: Department of Physiology, Faculty of Medicine, The University of Hong Kong.
authors.forename: J Z
authors.initials: JZ
authors.lastname: Sheng
authors.forename: N S
authors.initials: NS
authors.lastname: Wong
authors.forename: H X
authors.initials: HX
authors.lastname: Wang
authors.forename: T M
authors.initials: TM
authors.lastname: Wong
mesh_terms: D019900:3,4-Dichloro-N-methyl-N-(2-(1-pyrrolidinyl)-cyclohexyl)-benzeneacetamide, (trans)-Isomer; D000818:Animals; D002118:Calcium / Q000378:metabolism*; D002469:Cell Separation; D003600:Cytosol / Q000378:metabolism*; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D004963:Estrenes / Q000494:pharmacology; D006352:Heart Ventricles; D008297:Male; D009206:Myocardium / Q000166:cytology / Q000378:metabolism*; D009994:Osmolar Concentration; D037342:Pertussis Toxin; D011505:Protein-Tyrosine Kinases / Q000037:antagonists & inhibitors*; D011759:Pyrrolidines / Q000037:antagonists & inhibitors / Q000494:pharmacology*; D011760:Pyrrolidinones / Q000494:pharmacology; D051381:Rats; D017207:Rats, Sprague-Dawley; D010566:Virulence Factors, Bordetella / Q000494:pharmacology*
chemical_list: D004791:Enzyme Inhibitors; D004963:Estrenes; D011759:Pyrrolidines; D011760:Pyrrolidinones; D010566:Virulence Factors, Bordetella; C060229:1-(6-((3-methoxyestra-1,3,5(10)-trien-17-yl)amino)hexyl)-1H-pyrrole-2,5-dione; D019900:3,4-Dichloro-N-methyl-N-(2-(1-pyrrolidinyl)-cyclohexyl)-benzeneacetamide, (trans)-Isomer; D037342:Pertussis Toxin; D011505:Protein-Tyrosine Kinases; D002118:Calcium