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Evidence of muscarinic cholinergic receptor in dog portal vein binding of tritium labeled quinuclidinyl benzilate

, : Evidence of muscarinic cholinergic receptor in dog portal vein binding of tritium labeled quinuclidinyl benzilate. Life Sciences 32(15): 1757-1764

Muscarinic cholinergic receptor sites in dog portal veins were analyzed directly using [3H]quinuclidinyl benzilate (QNB) as a ligand. Specific [3H]QNB binding to crude membrane preparations from the isolated veins was saturable, reversible and of high affinity (Kd = 15.5 .+-. 2.8 pM) with a Bmax [maximum bound concentration] of 110 .+-. 14.7 f[femto]mol/mg protein. Scatchard and Hill plot analyses of the data indicated 1 class of binding sites. From kinetic analysis of the data, association and dissociation rate constants of 1.91 .times. 109 M-1 min-1 and 0.016 min-1, respectively, were calculated. Kd calculated from the equation Kd = K-1/K+1 was 8.3 pM, in good agreement with the Scatchard estimate of Kd (15.5 pM). Specific binding of [3H]QNB was displaced by muscarinic agents. Nicotinic cholinergic agents, .alpha.-bungarotoxin, nicotine and hexamethonium, were ineffective in displacing [3H]QNB binding at 10 .mu.M. These findings provide direct evidence for the existence of muscarinic cholinergic receptors in dog portal veins.

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