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The polarity of cell-surface membrane movements and their regulation by adrenal steroid hormones (10(-6) M aldosterone) and vasopressin or vasotocin were studied in A6 cells. This cell line is derived from the Xenopus laevis distal nephron and displays regulated Na+ reabsorption but is devoid of regulated water transport. Apical and basolateral membrane movements and their hormonal regulation were characterized by measuring the uptake of the fluid phase marker horseradish peroxidase (HRP) and the secretion of proteins on both sides of cell monolayers cultured on filters. The intracellular accumulation of HRP was visualized by electron microscopy and quantified by the measure of cell-associated peroxidase activity. The rate of intracellular HRP accumulation corresponded to 0.01 nl/minute/filter (4.7 cm2) from the apical side and was 20-32 times faster from the basolateral side. In contrast, the level of protein secretion was 3.5 times higher apically than basolaterally. Among the secreted proteins some were found to be secreted essentially apically, and others basolaterally. Vasotocin increased apical endocytosis (1.88-fold) and apical protein secretion (1.49-fold) in cells pretreated with aldosterone. Basolaterally, only the endocytosis was increased, and to a smaller extent (1.36-fold). These effects of vasotocin depended on aldosterone pretreatment and could be mimicked with forskolin and 8-bromoadenosine 3':5'-cyclic monophosphate (BrcAMP). Measurements of intracellular cAMP levels showed that there was a rankorder correlation between the induced level of intracellular cAMP and that of apical endocytosis. This study shows that vasotocin has a polarized stimulatory action on apical endocytosis and protein secretion in A6 cells, and that the mediation of this action by cAMP is aldosterone dependent.
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