Carbachol-induced oxygen consumption in slices from developing rat submandibular and parotid glands.

In contrast to the submandibular gland, the developing rat parotid gland shows refractoriness to cholinergic secretagogues until 2 wks of age. To assess the underlining mechanism of this refractoriness, I investigated changes in oxygen consumption as a function of animal age in slices from rat submandibular and parotid glands, measuring both basal and carbachol-stimulated levels. The oxygen consumption was determined by a direct manometric method in the Warburg apparatus. Carbachol-induced oxygen uptake in submandibular gland slices was observed by 1 day of age and reached the adult level of stimulation by 3 wks of age. In the parotid gland, carbachol failed to stimulate oxygen uptake in the early post-natal period, and the first response was detected at 2 wks of age, reaching the adult level at 4 wks of age. Para-fluorohexahydro-sila-diphenidol (pFHHSiD), a selective M3 antagonist, inhibited carbachol-induced oxygen uptake in both glands, while pirenzepine, a selective M1 antagonist, had no effect, suggesting that the M3 muscarinic receptors are involved in this process. The respiratory effect of carbachol, in both glands, was inhibitable by ouabain and, to a lesser extent, by furosemide, indicating that carbachol-enhanced oxygen uptake is due to Na,K-ATPase and that the furosemide-sensitive co-transport of Na+ entry is underdeveloped in immature cells. The ouabain-sensitive Na,K-ATPase activity in the parotid gland increased from birth until 28 days of age. At the time of parotid gland refractoriness to carbachol, Ca2+ ionophore A23187 caused an increase of oxygen uptake only in the presence of extracellular Ca2+. In the presence of carbachol, the effect of ionophore was significantly higher than that of ionophore alone. These results raise the possibility that the refractoriness of the parotid gland to carbachol is due to the inability of carbachol to increase Ca2+ uptake rather than to the lack of distal limb, which resides on the pathway from receptor stimulation to Na,K-ATPase activation.