Corticotropin-releasing factor increases [Ca2+]i via receptor-mediated Ca2+ channels in human epidermoid A-431 cells.
Corticotropin-releasing factor (CRF) has been shown to attenuate vascular leakage in injured skin, mucous membrane, muscle, and brain. Calcium is thought to play an important role in many of the physiological responses to CRF, but there has been little characterization of how calcium is involved in process by which CRF protects damaged tissues. The goal of this study was to characterize changes in cytosolic free calcium concentrations ([Ca2+]i) in human epidermoid A-431 cells exposed to human/rat-CRF and to investigate the mechanisms by which these changes occur. The resting [Ca2+]i in normal cells at 37 degrees C was 66 +/- 4 nM (n = 32). When cells were treated with CRF, [Ca2+]i increased immediately. The increase depended on CRF concentration, with a median effective concentration of 11 pM. This increase in [Ca2+]i depended on external Ca2+ but not Na+, Mg2+, or K+. La3+ (10 microM) and Co2+ (10 microM) inhibited the CRF-induced [Ca2+]i increase, whereas verapamil and nifedipine tested at concentrations up to 1 mM did not. alpha-Helical CRF-(9-41), a synthetic CRF receptor antagonist, and pertussis toxin blocked the increase in [Ca2+]i induced by CRF, which suggests that the entry of extracellular Ca2+ is mediated by receptor-operated Ca2+ channels coupled with pertussis toxin-sensitive G proteins. Although 420 pM CRF stimulated an immediate increase in [Ca2+]i, inositol trisphosphate and cellular cAMP levels did not change within 1 min either in the presence or absence of external Ca2+.(ABSTRACT TRUNCATED AT 250 WORDS)
Kiang
J G
JG
Department of Clinical Physiology, Walter Reed Army Institute of Research, Washington, DC 20307-5100.
D000818:Animals; D002118:Calcium / Q000378:metabolism*; D002121:Calcium Channel Blockers / Q000494:pharmacology; D015220:Calcium Channels / Q000187:drug effects / Q000378:metabolism*; D002478:Cells, Cultured; D003346:Corticotropin-Releasing Hormone / Q000494:pharmacology*; D000242:Cyclic AMP / Q000378:metabolism; D006801:Humans; D015544:Inositol 1,4,5-Trisphosphate / Q000378:metabolism; D051379:Mice; D010446:Peptide Fragments / Q000494:pharmacology; D037342:Pertussis Toxin; D011863:Radioimmunoassay; D018019:Receptors, Corticotropin-Releasing Hormone / Q000037:antagonists & inhibitors / Q000187:drug effects*; D012867:Skin / Q000166:cytology / Q000378:metabolism*; D010566:Virulence Factors, Bordetella / Q000494:pharmacology
D002121:Calcium Channel Blockers; D015220:Calcium Channels; D010446:Peptide Fragments; D018019:Receptors, Corticotropin-Releasing Hormone; D010566:Virulence Factors, Bordetella; D015544:Inositol 1,4,5-Trisphosphate; D003346:Corticotropin-Releasing Hormone; C045089:corticotropin releasing hormone (9-41); D000242:Cyclic AMP; D037342:Pertussis Toxin; D002118:Calcium