Rapid effects of corticosteroids on cytosolic protein kinase C and intracellular calcium concentration in human distal colon.
Recent studies from our laboratory have reported rapid (< 1 min) non-genomic activation of potassium recycling, Na+-H+ exchange, protein kinase C (PKC) activity and PKC-sensitive Ca2+ entry by mineralocorticoids in mammalian distal colonic epithelium. Previous studies from other laboratories have described stimulation of the Na+-H+ exchanger by PKC activation. Here a rapid non-genomic effect of aldosterone on PKC activity and intracellular free calcium [Ca2+]i is demonstrated in human distal colonic epithelium. Rapid activation (after 15 min incubation) of basal PKC activity was observed in cytosolic fractions of human colonic epithelium by aldosterone, fludrocortisone and deoxycorticosterone acetate (DOCA). PKC activation was inhibited by the specific PKC inhibitor bisindolylmaleimide (GF109203X). The glucocorticoid hydrocortisone failed to activate PKC activity. Aldosterone induced a rapid increase in [Ca2+]i in isolated human colonic crypts. This stimulatory effect on [Ca2+]i was inhibited by the PKC inhibitor chelerythrine chloride. Hydrocortisone and dexamethasone similarly failed to increase [Ca2+]i. These results indicate that intracellular signalling for aldosterone involves changes in [Ca2+]i via activation of PKC. Since stimulation of PKC activity and increase in [Ca2+]i are apparent at normal circulating levels of aldosterone, our findings may have important physiological implications and prompt a reassessment of mineralocorticoid effects on electrolyte homeostasis.
Doolan
C M
CM
Wellcome Trust Cellular Physiology Research Unit, Department of Physiology, University College, Mercy Hospital, Cork, Ireland. stpy8004@bureau.ucc.ie
O'Sullivan
G C
GC
Harvey
B J
BJ
D000305:Adrenal Cortex Hormones / Q000494:pharmacology*; D000450:Aldosterone / Q000494:pharmacology; D002118:Calcium / Q000378:metabolism*; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism; D003106:Colon / Q000378:metabolism*; D003600:Cytosol / Q000201:enzymology; D003900:Desoxycorticosterone / Q000494:pharmacology; D004789:Enzyme Activation; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005438:Fludrocortisone / Q000494:pharmacology; D006801:Humans; D006854:Hydrocortisone / Q000494:pharmacology; D066298:In Vitro Techniques; D007211:Indoles / Q000494:pharmacology; D007413:Intestinal Mucosa / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D007700:Kinetics; D008301:Maleimides / Q000494:pharmacology; D008901:Mineralocorticoids / Q000494:pharmacology / Q000502:physiology; D011493:Protein Kinase C / Q000378:metabolism*; D013997:Time Factors
D000305:Adrenal Cortex Hormones; D004791:Enzyme Inhibitors; D007211:Indoles; D008301:Maleimides; D008901:Mineralocorticoids; D003900:Desoxycorticosterone; D000450:Aldosterone; D011493:Protein Kinase C; C070515:bisindolylmaleimide I; D002118:Calcium; D005438:Fludrocortisone; D006854:Hydrocortisone