Modulation of cardiac gap junctions: the mode of action of arachidonic acid.
Myocytes isolated from neonatal rat hearts were grown in culture dishes. Cell pairs were selected to examine the mode of action of arachidonic acid (AA) on gap junctions. The dual voltage-clamp method was used to measure intercellular currents and determine the gap junction conductance, gj. Exposure of cell pairs to 10 microM AA produced reversible uncoupling. Pretreatment with 10 microM POCA (sodium-2-[5-(4-chlorophenyl)-pentyl]-oxirane-2-carboxylate; which inhibits mitochondrial beta-oxidation) did not prevent AA-dependent uncoupling. Thus, it seems that metabolites of beta-oxidation are not involved in AA-induced impairment of gj. Pre-exposure to 10 microM indomethacin (which blocks the cyclooxygenase pathway of the AA-cascade) had no effect on AA-dependent uncoupling. This suggests that cyclooxygenase products such as prostaglandins or thromboxanes play no role in gj modulation. Exposure to 5 microM NDGA (nordihydroguaiaretic acid; which inhibits the 5-lipoxygenase pathway) or 10 microM ETYA (5,8,11,14-eicosatetrynoic acid: which inhibits the 12- and 15-lipoxygenase pathway) led to a reversible decrease in gj. Pre-treatment with 4-BPB (4-bromophenacyl bromide: which inhibits phospholipase A2) did not prevent the effects on gj by NDGA or ETYA. This renders it unlikely that gj is regulated by eicosanoids. Also, accumulation of endogenous AA cannot be responsible for NDGA- and ETYA-dependent uncoupling. Exposure to 75 microM SKF-525A (inhibits the epoxygenase pathway) reversibly impaired gj. This is consistent with a direct action of SKF-525A on gj, but leaves open the possibility of an involvement of epoxides. The data gathered will be discussed in terms of molecular mechanisms. Due to their amphipathic character. AA, NDGA, ETYA and SKF-525A may interfere with gj by disturbing the lipid-protein interface of the cell membranes and thereby impair gap junction channels.
Schmilinsky-Fluri
G
G
Department of Physiology, University of Bern, Switzerland.
Valiunas
V
V
Willi
M
M
Weingart
R
R
D000818:Animals; D016718:Arachidonic Acid / Q000378:metabolism*; D002478:Cells, Cultured; D016861:Cyclooxygenase Inhibitors / Q000494:pharmacology; D000090762:Cytochrome P-450 CYP2J2; D003577:Cytochrome P-450 Enzyme System / Q000187:drug effects / Q000378:metabolism; D015777:Eicosanoids / Q000378:metabolism; D004852:Epoxy Compounds / Q000494:pharmacology; D017629:Gap Junctions / Q000187:drug effects / Q000378:metabolism*; D007004:Hypoglycemic Agents / Q000494:pharmacology; D007213:Indomethacin / Q000494:pharmacology; D008084:Lipoxygenase / Q000187:drug effects / Q000378:metabolism; D016859:Lipoxygenase Inhibitors / Q000494:pharmacology; D009637:Masoprocol / Q000494:pharmacology; D009206:Myocardium / Q000166:cytology / Q000378:metabolism*; D010084:Oxidation-Reduction; D010105:Oxygenases / Q000187:drug effects / Q000378:metabolism; D011451:Prostaglandin-Endoperoxide Synthases / Q000187:drug effects / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar
D016861:Cyclooxygenase Inhibitors; D015777:Eicosanoids; D004852:Epoxy Compounds; D007004:Hypoglycemic Agents; D016859:Lipoxygenase Inhibitors; D016718:Arachidonic Acid; D009637:Masoprocol; C033891:ethyl 2-(5-(4-chlorophenyl)pentyl)oxiran-2-carboxylate; D003577:Cytochrome P-450 Enzyme System; D010105:Oxygenases; D008084:Lipoxygenase; D000090762:Cytochrome P-450 CYP2J2; D011451:Prostaglandin-Endoperoxide Synthases; D007213:Indomethacin