title: 5-Lipoxygenase metabolites of arachidonic acid regulate volume decrease by mudpuppy red blood cells.
abstract: We examined whether metabolites of arachidonic acid (AA) regulate K+ efflux during regulatory volume decrease (RVD) by mudpuppy red blood cells (RBCs). Volume regulation was inhibited by the phospholipase A2 antagonists mepacrine (10 microM) and ONO-RS-082 (10 microM); the inhibitory effect of ONO-RS-082 was reversed by gramicidin (5 microM). Eicosatetraynoic acid (ETYA, 100 microM), a general antagonist of AA metabolism, also blocked RVD. In addition, volume regulation was inhibited by the lipoxygenase pathway antagonist nordihydroguaiaretic acid (NDGA, 10 microM), the 5 lipoxygenase antagonists AA-861 (5 microM) and curcumin (20 microM), and by the 5-lipoxygenase activating protein inhibitor L-655,298 (5 microM). Inhibition by all four of these agents was reversed with gramicidin. In contrast, the 12- and 15-lipoxygenase pathway inhibitor ethyl-3,4-dihydroxy-benzylidene-cyanoacetate (EDBCA, 1 microM) and the cytochrome P-450 monooxygenase pathway blocker ketoconazole (20 microM) had no effect. On the other hand, the cyclooxygenase pathway inhibitor aspirin (100 microM) slightly enhanced RVD. Consistent with these findings, a K(+)-selective whole cell conductance responsible for K+ efflux during cell swelling was inhibited by ONO-RS-082 (10 microM), NDGA (10 microM), AA-861 (5 microM), curcumin (20 microM), and L-655,298 (5 microM). In contrast, EDBCA (1 microM), ketoconazole (20 microM), and indomethacin (10 microM) did not block this whole cell conductance. These results indicate that a channel mediating K+ loss during RVD is regulated by a 5-lipoxygenase metabolite of arachidonic acid.
authors.affiliation: Department of Biology, Ripon College, WI 54971, USA.
authors.forename: D B
authors.initials: DB
authors.lastname: Light
authors.forename: T M
authors.initials: TM
authors.lastname: Mertins
authors.forename: J A
authors.initials: JA
authors.lastname: Belongia
authors.forename: C A
authors.initials: CA
authors.lastname: Witt
mesh_terms: D000818:Animals; D001094:Arachidonate 5-Lipoxygenase / Q000097:blood*; D016718:Arachidonic Acid / Q000097:blood*; D048429:Cell Size; D004912:Erythrocytes / Q000378:metabolism / Q000502:physiology*; D006400:Hematocrit; D009337:Necturus; D009996:Osmotic Fragility; D018408:Patch-Clamp Techniques
chemical_list: D016718:Arachidonic Acid; D001094:Arachidonate 5-Lipoxygenase