Arachidonic acid activates cation channels in bovine adrenal chromaffin cells.
Microscopic fluorescence analysis of fura-2-loaded bovine adrenal chromaffin cells demonstrates that approximately 70% of the cells responded to arachidonic acid in increasing the intracellular Ca2+ concentration. Because this increase was markedly less in the absence of external Ca2+, we examined the effect of arachidonic acid on Ca2+ influx electrophysiologically. Bath application of 10 microM arachidonic acid induced a long-lasting inward current when the cell was clamped at -50 mV. Other fatty acids, such as oleic acid, linoleic acid, eicosatrienoic acid, and eicosapentaenoic acid, were all ineffective. The current-voltage relationships suggest that arachidonic acid may activate voltage-insensitive channels. Arachidonic acid (> or = 2 microM) activated a single-channel current in the inside-out patch, even in the presence of inhibitors of cyclooxygenase and lipoxygenase, possibly suggesting that arachidonic acid could activate channels directly. The onset delay of the inward channel current in the outside-out patch configuration (54.2 +/- 63.5 s; mean +/- SD) was significantly shorter than that in the inside-out patch one (197.3 +/- 177.7 s). Washout of arachidonic acid decreased the probability of channel openings in the outside-out patch but not in the inside-out one. These results suggest that arachidonic acid activates channels reversibly from outside of the plasma membrane. The unitary conductance for Ca2+ of arachidonic acid-activated channel was approximately 17 pS. The arachidonic acid-activated channel was permeable to Ba2+, Ca2+, and Na+ but not to Cl-. The opening probability of the arachidonic acid-activated channel did not depend on membrane potential.(ABSTRACT TRUNCATED AT 250 WORDS)
Mochizuki-Oda
N
N
Department of Cell Biology and Neuroscience, Osaka Bioscience Institute, Japan.
Negishi
M
M
Mori
K
K
Ito
S
S
D000313:Adrenal Medulla / Q000187:drug effects / Q000378:metabolism / Q000502:physiology*; D000818:Animals; D016718:Arachidonic Acid / Q000494:pharmacology*; D002118:Calcium / Q000378:metabolism*; D002413:Cations, Divalent / Q000378:metabolism; D002417:Cattle; D002478:Cells, Cultured; D015232:Dinoprostone / Q000494:pharmacology; D004594:Electrophysiology / Q000379:methods; D007473:Ion Channels / Q000187:drug effects / Q000502:physiology*; D015289:Leukotrienes / Q000494:pharmacology; D008054:Lipid Peroxides / Q000494:pharmacology; D008564:Membrane Potentials / Q000187:drug effects; D013779:Tetrodotoxin / Q000494:pharmacology
D002413:Cations, Divalent; D007473:Ion Channels; D015289:Leukotrienes; D008054:Lipid Peroxides; D016718:Arachidonic Acid; D013779:Tetrodotoxin; C021201:12-HPETE; C025086:15-hydroperoxy-5,8,11,13-eicosatetraenoic acid; D015232:Dinoprostone; D002118:Calcium