Discovery of 5R-lipoxygenase activity in oocytes of the surf clam, Spisula solidissima.
Arachidonic acid and 5-hydroxyeicosatetraenoic acid (5-HETE) are reported to induce reinitiation of meiosis in oocytes of the surf clam Spisula sachalinensis from the Sea of Japan (Varaksin et al., Comp. Biochem. Physiol. 101C, 627-630 (1992). As the Atlantic surf clam Spisula solidissima is a commonly used model for the study of meiosis reinitiation, we examined these cells for the possible occurrence of lipoxygenases and for the bioactivity of the products. Incubation of [14C]arachidonic acid with homogenates of S. solidissima oocytes led to the formation of two major metabolites: 5R-HETE, a novel lipoxygenase product, and 8R-HETE. The products were identified by HPLC, uv spectroscopy, and GC-MS. The corresponding hydroperoxy fatty acids, the primary lipoxygenase products, were isolated from incubations of ammonium sulfate fractionated oocyte cytosol. Arachidonic and eicosapentaenoic acids were identified as constituents of S. solidissima oocyte lipids and the free acids were equally good lipoxygenase substrates. We examined the activity of C18 and C20 polyunsaturated fatty acids and their lipoxygenase products on meiosis reinitiation in Spisula solidissima oocytes, using serotonin and ionophore A23187 as positive controls. The fatty acids and their derivatives were inactive. We conclude that in the surf clam, (as in starfish), there are responding and non-responding species in regard to the maturation-inducing activity of the oocyte lipoxygenase products, and that the lipoxygenase has another, as yet uncharacterized, function in oocyte physiology.
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Department of Pharmacology, Vanderbilt University Medical Center, Nashville, TN 37232-6602, USA.
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Brash
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D000818:Animals; D001094:Arachidonate 5-Lipoxygenase / Q000378:metabolism*; D016718:Arachidonic Acid / Q000378:metabolism; D049872:Bivalvia / Q000201:enzymology*; D000001:Calcimycin / Q000494:pharmacology; D002122:Calcium Chloride / Q000494:pharmacology; D002458:Cell Fractionation; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange; D015118:Eicosapentaenoic Acid / Q000031:analogs & derivatives / Q000378:metabolism; D004789:Enzyme Activation / Q000187:drug effects; D005227:Fatty Acids / Q000032:analysis; D006893:Hydroxyeicosatetraenoic Acids / Q000378:metabolism; D008968:Molecular Conformation; D009865:Oocytes / Q000737:chemistry / Q000201:enzymology*; D012701:Serotonin / Q000494:pharmacology; D013053:Spectrophotometry
D005227:Fatty Acids; D006893:Hydroxyeicosatetraenoic Acids; D016718:Arachidonic Acid; D012701:Serotonin; D000001:Calcimycin; C022022:5-hydroxy-6,8,11,14-eicosatetraenoic acid; C047628:8-hydroxyeicosatetraenoic acid; D015118:Eicosapentaenoic Acid; D001094:Arachidonate 5-Lipoxygenase; D002122:Calcium Chloride