Muscarinic receptor coupling to inositol phospholipid metabolism in guinea-pig cerebral cortex, parotid gland and ileal smooth muscle.
Inositol phospholipid hydrolysis induced by agonist-stimulation of muscarinic receptors has been examined in slices of guinea-pig cerebral cortex, parotid gland and ileal smooth muscle. An assay measuring 3H-inositol phosphate formation from prelabelled lipids in the presence of LiCl, allowed marked stimulation by agonists to be followed. The pD2-value of carbachol differed markedly, between tissues being more than 10-fold lower in cerebral cortex than in parotid gland. The partial agonist oxotremorine showed the largest relative maximal responsiveness in parotid gland, followed by ileum and cortex. Atropine suppressed the phosphoinositide response to carbachol with an almost similar affinity in each tissue, but pirenzepine was found to have a 20-fold higher affinity in cerebral cortex, pKi = 7.7 than in parotid gland, pKi = 6.3. Carbachol, even in the presence of guanosine triphosphate (GTP), displayed complex binding against 3H-N-methylscopolamine (3H-NMS) in cortical and ileal membranes, though in membranes from the parotid gland a single homogeneous population was found. Atropine inhibition of 3H-NMS parallelled its suppression of the phosphoinositide response, the affinities in each tissue studied being similar. Pirenzepine inhibited binding from two components in cerebral cortex, the high affinity value being similar to that obtained in the phosphoinositide assay. In parotid gland, however, only low affinity pirenzepine binding sites were observed, closely resembling the affinity found for this antagonist in the functional assay. These experiments suggest (a) that there are differences between agonist occupation of muscarinic receptors and phosphoinositide hydrolysis within the different tissues, (b) that both high and low affinity pirenzepine binding sites appear to be linked to phosphoinositide metabolism, and (c) that low affinity pirenzepine sites may be more efficiently coupled to the hydrolysis of phosphoinositides.
Ek
B
B
Department of Pharmacology and Therapeutics, University of Leicester, U.K.
Nahorski
S
S
D000818:Animals; D001285:Atropine / Q000494:pharmacology; D002217:Carbachol / Q000493:pharmacokinetics; D002540:Cerebral Cortex / Q000378:metabolism*; D006160:Guanosine Triphosphate / Q000494:pharmacology; D006168:Guinea Pigs; D007082:Ileum; D009130:Muscle, Smooth / Q000378:metabolism*; D019832:N-Methylscopolamine; D010095:Oxotremorine / Q000493:pharmacokinetics; D010306:Parotid Gland / Q000378:metabolism*; D010716:Phosphatidylinositols / Q000378:metabolism*; D010890:Pirenzepine / Q000493:pharmacokinetics; D011976:Receptors, Muscarinic / Q000378:metabolism*; D012602:Scopolamine Derivatives / Q000494:pharmacology
D010716:Phosphatidylinositols; D011976:Receptors, Muscarinic; D012602:Scopolamine Derivatives; D010890:Pirenzepine; D010095:Oxotremorine; D001285:Atropine; D006160:Guanosine Triphosphate; D002217:Carbachol; D019832:N-Methylscopolamine