Dispersed acini isolated by collagenase digestion of the rat submandibular gland were used to compare the effects of amiloride and furosemide on the uptake of the isotopic tracer 22Na and on the binding of [3H]quinuclidinyl benzylate ([3H]QNB). In mM concentrations, both inhibitors reduced 22Na uptake in resting cells 34 and 25-29%, respectively. Acetylcholine (1 microM) enhanced uptake 23% and this effect was reduced 45% by amiloride and 26% by furosemide. Amiloride inhibited the binding of [3H]QNB to crude membranes prepared from fresh submandibular glands in a dose-dependent fashion (IC50 = 8 x 10(-6) M). Furosemide (3 x 10(-8) to 10(-3) M) did not inhibit radioligand binding. Na influx into resting salivary acini thus appears to occur by both amiloride-sensitive and furosemide-sensitive transport systems. The similar inhibition by furosemide of unstimulated and stimulated uptake of 22Na suggests that acetylcholine does not significantly activate the cotransport system within the time frame (i.e., 2 min) of the experiments. Acetylcholine appears to activate an amiloride-sensitive Na/H antiport, but amiloride blocks cholinergic receptors and may thus affect Na transport by receptor blockade. Other actions of amiloride, such as its ability to penetrate into cells and to act as a weak base which alters intracellular pH, may also contribute to the inhibition of Na entry into salivary cells.
Acetylcholine Amiloride