Ca2+ release and contraction induced by IP3 and contractile agonists in mammalian gastric smooth muscle.
The source, time course and stoichiometry of cytosolic free Ca2+ ([Ca2+]i) during contraction were examined in smooth muscle cells isolated from the guinea pig and human stomach. Contraction by receptor-linked agonists (eg, acetylcholine, cholecystokinin octapeptide and Met-enkephalin) was preceded by stoichiometric increases in [Ca2+]i and net 45Ca2+ efflux that were maintained in the absence of extracellular Ca2+ or in the presence of a Ca2+ channel blocker (D600). The intracellular Ca2+ store could be depleted by repeated stimulation with all agonists in Ca2+-free medium or in the presence of D600 resulting in loss of contractile response; response was restored by re-exposure of the cells to Ca2+. The source of intracellular Ca2+ an the signal for its release were examined in saponin-permeabilized muscle cells. The cells retained their ability to contract in response to receptor-linked agonists and developed an ability to contract in response to inositol trisphosphate (IP3). The cells accumulated Ca2+ to the same extent as intact muscle cells, but only in the presence of ATP. IP3 caused a prompt, concentration-dependent increase in contraction, [Ca2+]i and net 45Ca2+ efflux. These effects were maximally similar to those produced by CCK-8 alone or in combination with IP3. Depletion of the Ca2+ store by repeated stimulation of single muscle cells in Ca2+-free medium with IP3, acetylcholine or CCK-8 separately resulted in loss of contractile response to all three agents; the response was restored by re-exposure of the muscle cell to cytosol-like perfusate (Ca2+ 180 nM).(ABSTRACT TRUNCATED AT 250 WORDS)
Makhlouf
G M
GM
Department of Medicine, Medical College of Virginia, Richmond 23298-0711.
D000109:Acetylcholine / Q000494:pharmacology; D000818:Animals; D002118:Calcium / Q000378:metabolism*; D003600:Cytosol / Q000378:metabolism; D004744:Enkephalin, Methionine / Q000494:pharmacology; D005711:Gallopamil / Q000494:pharmacology; D006168:Guinea Pigs; D006801:Humans; D066298:In Vitro Techniques; D015544:Inositol 1,4,5-Trisphosphate; D007295:Inositol Phosphates / Q000494:pharmacology*; D009119:Muscle Contraction / Q000187:drug effects; D009130:Muscle, Smooth / Q000187:drug effects*; D012844:Sincalide / Q000494:pharmacology; D013270:Stomach; D013403:Sugar Phosphates / Q000494:pharmacology*
D007295:Inositol Phosphates; D013403:Sugar Phosphates; D005711:Gallopamil; D004744:Enkephalin, Methionine; D015544:Inositol 1,4,5-Trisphosphate; D012844:Sincalide; D000109:Acetylcholine; D002118:Calcium