Characteristics of response of isolated smooth muscle cells to cholinergic drugs

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Abstract
The contractile responses of suspensions of isolated smooth muscle cells from the stomach muscularis of Bufo marinus were assessed with a Coulter counter. Contractile responses of strips from the same tissue were recorded isotonically. Suspensions of isolated smooth muscle cells exhibit a dose-dependent graded response to cholinergic agonists. The intact tissue has a lower sensitivity than the isolated cells to acetycholine but a higher sensitivity to carbachol. The higher sensitivity of isolated cells to acetycholine probably reflected decreased efficiency of cholinesterases upon removal of diffusion barriers. Because the isolated smooth muscle cells have affinity constants for atropine (K1 equals 0.07 +/- 0.02NM) and carbachol (9.5 +/- 3.7 muM) similar to that in this and other intact tissues, the affinity of the cholinergic receptor appears unaffected by cell isolation. The rate constant for dissociation (k2) of atropine was estimated from the slowing of response to carbachol by atropine; k2 in the isolated cells is 100 times larger than seen in intact tissues. Further insight into the interaction of cholinergic substances with their smooth muscle receptors might by obtained using this system. The isolated cells contain intact receptors, and diffusion limitations inherent to intact tissue have been removed.

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