Intracellular signals coupled to different rat ileal muscarinic receptor subtypes.
Taking into account that the activation of different subtypes of ileal muscarinic acetylcholine receptors (mAChR) regulate gut functions such as tone, motility, and electrolyte secretion, we characterized the expression of mAChR in ileal-purified membranes. We also studied intracellular signals triggered by mAChR activation. Binding parameters obtained from saturation assays with the nonselective tritiated muscarinic antagonist, quinuclidynil benzilate ([3H]-QNB), were maximal number of binding sites (Bmax): 30 +/- 2 fmol/mg prot and dissociation constant (Kd): 0.2 +/- 0.03 nM. The competitive inhibition of [3H]-QNB specific binding by various nonlabelled muscarinic antagonists was measured and the rank order of potency was: atropine (ATROP) > 4-DAMP > AF-DX 116 > pirenzepine (PZ). The activation of mAChR by carbachol (CARB) increased ileal motility in a concentration-dependent manner (EC50 2 x 10[-7] M). The antagonists' order of potency to displace dose-response curve of CARB was: ATROP > 4-DAMP > AF-DX116 > PZ. Optimal concentration of CARB on ileal strips increased phosphoinositide turnover and cGMP levels by activating ml receptor subtype and decreased isoproterenol (ISO) stimulated levels of cAMP due to M2 receptor activation. We can conclude that the activation of different mAchR subtypes triggers different intracellular signals that could regulate intestinal tone and motility.
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Centro de Estudios Farmacológicos y Botánicos, Consejo Nacional de Investigaciones Científicas y Técnicas, School of Dentistry, University of Buenos Aires, Argentina.
Sterin-Borda
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Rodriguez
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D000818:Animals; D002462:Cell Membrane / Q000378:metabolism; D000242:Cyclic AMP; D006152:Cyclic GMP; D004789:Enzyme Activation; D007082:Ileum / Q000166:cytology / Q000378:metabolism*; D066298:In Vitro Techniques; D007295:Inositol Phosphates; D007700:Kinetics; D018727:Muscarinic Antagonists / Q000494:pharmacology; D011813:Quinuclidinyl Benzilate / Q000494:pharmacology; D051381:Rats; D011976:Receptors, Muscarinic / Q000096:biosynthesis / Q000378:metabolism*; D012044:Regression Analysis; D015398:Signal Transduction / Q000502:physiology
D007295:Inositol Phosphates; D018727:Muscarinic Antagonists; D011976:Receptors, Muscarinic; D011813:Quinuclidinyl Benzilate; D000242:Cyclic AMP; D006152:Cyclic GMP