Effects of high-energy phosphates on carbachol-evoked cationic current in single smooth muscle cells from guinea-pig ileum.
1. Single smooth muscle cells from the longitudinal muscle layer of guinea-pig small intestine were voltage clamped in the whole-cell recording mode with patch pipettes. The cationic current (Icat) evoked by application of 50 microM carbachol (CCh) was examined when free internal calcium in the cell was 'clamped' at 10(-7) M with 20 mM BAPTA. The effects of varying the composition of the pipette solution were studied. 2. Phosphocreatine (PCr, 6 mM) added to the pipette solution increased Icat by about 7-fold (to near 620 pA); lower concentrations had similar, generally lesser, effects. Na2ATP (3 or 6 mM) with or without 5 mM MgCl2 was much less effective than phosphocreatine alone. Addition of 3 mM Na2ATP reduced Icat, whether or not phosphocreatine was present. 3. Creatine (6 mM) with or without 2 mM Na2ATP was less effective than phosphocreatine in maintaining Icat. 4. GTP (0.1 mM) did not affect Icat evoked by CCh, whether phosphocreatine was present or not. 5. GTP gamma S (0.2 mM) included in pipette solution mimicked the effect of CCH and evoked Icat independently of whether PCr was present or not in the pipette solution. Including 5 mM ATP in the pipette reduced this current, whereas 5'-adenylyl imidodiphosphate (AMP-PNP) and ADP were without effect. 6. The results show that phosphocreatine increases membrane channel responsiveness to receptor activation and that ATP above 2 mM suppresses it.
Bakhramov
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Department of Pharmacology and Clinical Pharmacology, St George's Hospital Medical School, London, UK.
D000255:Adenosine Triphosphate / Q000494:pharmacology*; D000818:Animals; D002217:Carbachol / Q000494:pharmacology*; D002412:Cations / Q000378:metabolism*; D002463:Cell Membrane Permeability; D006160:Guanosine Triphosphate / Q000494:pharmacology*; D006168:Guinea Pigs; D007082:Ileum; D066298:In Vitro Techniques; D007473:Ion Channels / Q000187:drug effects / Q000378:metabolism*; D008297:Male; D008564:Membrane Potentials; D009130:Muscle, Smooth / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D018408:Patch-Clamp Techniques; D010725:Phosphocreatine / Q000494:pharmacology*
D002412:Cations; D007473:Ion Channels; D010725:Phosphocreatine; D006160:Guanosine Triphosphate; D000255:Adenosine Triphosphate; D002217:Carbachol