Operation of Internal Na-dependent Ca Influx Mechanism Associated with Catecholamine Secretion in the Adrenal Chromatin Cells

Abstract

The cat adrenal gland was retrogradely perfused with a medium lacking divalent cation, and the secretion of catecholamines was induced by Ca reintroduction with or without simultaneous removal of external Na. These responses were markedly reduced after prior exposure to a medium containing Mg (1mM) or lacking Na. The inhibition caused by Mg was reversed by ouabain treatment. Reversing the concentration gradient for Na alone by replacing external Na with Tris, choline, or Li did not stimulate catecholamine secretion when Ca²⁺ (0.1mM) was present in the external medium throughout the experiment. However, the treatment with ouabain reversed this inhibitory effect of Ca so that Na removal did induce marked secretion. Essentially similar results concerning secretion were obtained in the cultured bovine chromaffin cells. There was a reasonable correlation between secretion and ⁴⁵Ca uptake in these cells under various experimental manipulations, and alterations of these parameters were well correlated with the level of internal Na. Furthermore, the rate constant of ²²Na efflux was found to increase when Ca was reintroduced with the simultaneous removal of Na during exposure to a medium containing ouabain. These results are consistent with the view that the mechanism of internal Na-dependent Ca influx is activated to induce catecholamine secretion whenever internal Na is raised above a critical level. On the other hand, there were significant increases in the catecholamine secretion and ⁴⁵Ca uptake which were induced by substitution of NaCl with sucrose, even when the operation of the internal Na-dependent Ca influx mechanism was markedly restricted by various experimental manipulations. This suggests that other secretory mechanisms are involved under these conditions. This would account for the largest secretory effect of sucrose substitution under the condition in which internal Na-dependent Ca influx is activated.