Incorporation of [3H]arachidonic and [14C]eicosapentaenoic acids into glycerophospholipids and their metabolism via lipoxygenases in isolated brain cells from rainbow trout Oncorhynchus mykiss.
The incorporation of [3H]arachidonate [( 3H]AA) and [14C]eicosapentaenoate [( 14C]EPA) into glycerophospholipids was studied in isolated brain cells from rainbow trout, a teleost fish whose lipids are rich in (n-3) polyunsaturated fatty acids (PUFAs). EPA was incorporated into total lipid to a greater extent than AA, but the incorporation of both PUFAs into total glycerophospholipids was almost identical. The incorporation of both AA and EPA was greatest into phosphatidylethanolamine (PE). However, when expressed per milligram of individual phosphoglycerides, both AA and EPA were preferentially incorporated into phosphatidylinositol (PI), the preference being significantly greater with AA. On the same basis, significantly more EPA than AA was incorporated into phosphatidylcholine (PC). When double-labelled cells were challenged with calcium ionophore A23187, the 3H and 14C released from the cells closely paralleled each other, peaking at 10 min after addition of ionophore. The 12-monohydroxylated derivative was the pre-dominant lipoxygenase product from both AA and EPA with a rank order of 12-hydroxyeicosatetraenoic acid (12-HETE) greater than leukotriene B4 (LTB4) greater than 5-HETE greater than 15-HETE for the AA products and 12-hydroxyeicosapentaenoic acid (12-HEPE) greater than 5-HEPE greater than LTB5 greater than 15 HEPE for EPA products. The 3H/14C (dpm/dpm) ratios in the glycerophospholipids, total released radioactivity, and the lipoxygenase products suggested that PC rather than PI was the likely source of eicosanoid precursors in trout brain cells.
Tocher
D R
DR
NERC Unit of Aquatic Biochemistry, School of Natural Sciences, University of Stirling, Scotland.
Bell
J G
JG
Sargent
J R
JR
D000818:Animals; D016718:Arachidonic Acid / Q000493:pharmacokinetics*; D001921:Brain / Q000166:cytology / Q000378:metabolism*; D000001:Calcimycin / Q000494:pharmacology; D002250:Carbon Radioisotopes; D002469:Cell Separation; D015118:Eicosapentaenoic Acid / Q000493:pharmacokinetics*; D006893:Hydroxyeicosatetraenoic Acids / Q000378:metabolism; D007975:Leukotriene B4 / Q000378:metabolism; D008084:Lipoxygenase / Q000378:metabolism*; D009474:Neurons / Q000378:metabolism; D010712:Phosphatidic Acids / Q000378:metabolism*; D014316:Tritium; D014337:Trout / Q000378:metabolism*
D002250:Carbon Radioisotopes; D006893:Hydroxyeicosatetraenoic Acids; D010712:Phosphatidic Acids; D014316:Tritium; D007975:Leukotriene B4; D016718:Arachidonic Acid; D000001:Calcimycin; D015118:Eicosapentaenoic Acid; D008084:Lipoxygenase