Modulation of cytosolic protein kinase C and calcium ion activity by steroid hormones in rat distal colon.
Studies from our laboratory have demonstrated rapid (<1 min) non-genomic activation of Na+-H+ exchange and potassium recycling by mineralocorticoids in human and rat colonic epithelium. It has previously been demonstrated that Na+-H+ exchange may be stimulated by protein kinase C (PKC) activation; therefore, we examined the effect of mineralocorticoids on PKC activity in rat colonic epithelium. Activation (after 15 min of incubation) of basal PKC activity was observed in cytosolic fractions of rat colonic epithelium by aldosterone, fludrocortisone, and deoxycorticosterone acetate. In all instances, PKC activation was inhibited by the PKC inhibitor bisindolylmaleimide (GF109203X). Hydrocortisone failed to activate PKC activity. Stimulation of basal intracellular free calcium [Ca2+]i was observed, in isolated rat colonic crypts, following aldosterone addition. This stimulatory effect was inhibited by the PKC inhibitor, chelerythrine chloride. Hydrocortisone failed to increase [Ca2+]i. These results indicate that intracellular signaling for aldosterone involves changes in [Ca2+]i via activation of PKC. Since the stimulation of PKC and increase in [Ca2+]i are apparent at normal circulating levels of aldosterone, our findings have major implications for the reassessment of mineralocorticoid effects on electrolyte homeostasis.
Doolan
C M
CM
Wellcome Trust Cellular Physiology Research Unit, Department of Physiology, University College, Cork, Ireland.
Harvey
B J
BJ
D000450:Aldosterone / Q000494:pharmacology*; D000818:Animals; D000893:Anti-Inflammatory Agents / Q000494:pharmacology; D002118:Calcium / Q000378:metabolism*; D002462:Cell Membrane / Q000201:enzymology; D003106:Colon / Q000378:metabolism*; D003600:Cytosol / Q000201:enzymology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D004958:Estradiol / Q000494:pharmacology; D006854:Hydrocortisone / Q000494:pharmacology; D007211:Indoles / Q000494:pharmacology; D007413:Intestinal Mucosa / Q000201:enzymology*; D008301:Maleimides / Q000494:pharmacology; D010718:Phosphatidylserines / Q000494:pharmacology; D011493:Protein Kinase C / Q000037:antagonists & inhibitors / Q000378:metabolism*; D051381:Rats
D000893:Anti-Inflammatory Agents; D004791:Enzyme Inhibitors; D007211:Indoles; D008301:Maleimides; D010718:Phosphatidylserines; D000450:Aldosterone; D004958:Estradiol; D011493:Protein Kinase C; C070515:bisindolylmaleimide I; D002118:Calcium; D006854:Hydrocortisone