Abstract
We characterized agonist-induced internalization, recycling and downregulation of each muscarinic receptor subtype (M 1–M 5) stably expressed in Chinese hamster ovary (CHO) cells. The radioligands [ 3H]QNB and [ 3H]NMS were used to measure the total and plasma membrane populations of muscarinic receptors, respectively. Following carbachol treatment (1 mM), the rank orders for the rate of carbachol-induced internalization of the muscarinic subtypes were M 2 > M 4 = M 5 > M 3 = M 1, respectively. Unlike the M 2 receptor, M 1, M 3, M 4 and M 5 receptors recycled back to the plasma membrane after 1 h carbachol treatment. The receptor downregulation elicited to 24 h carbachol treatment was similar for M 2, M 3, M 4 and M 5 receptors, whereas that for the M 1 receptor was greater. Our results indicate that there are subtype-specific differences in the rate and extent of agonist-induced muscarinic receptor internalization, recycling and downregulation in CHO cells.
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