Activation of phospholipase D by prostaglandin F2 alpha in rat luteal cells and effects of inhibitors of arachidonic acid metabolism.
In rat luteal cells labeled with [3H]oleic acid, PGF2 alpha-stimulated phospholipase D (PLD) activation was investigated. The PLD activity was detected by measuring the accumulation of [3H]phosphatidylethanol (PtdEt) in the presence of ethanol. PGF2 alpha stimulated PtdEt accumulation at concentrations of more than 100 nM in the presence of ethanol. However, PtdEt accumulation did not change in the absence of ethanol. PGF2 alpha (1 microM) increased PtdEt accumulation after 1 min, and the accumulation reached a plateau by 2-3 min. These results indicate that PGF2 alpha activates PLD in rat luteal cells. U-73122, a phospholipase C (PLC) inhibitor, and staurosporine, a protein kinase C (PKC) inhibitor, did not inhibit PGF2 alpha-stimulated [3H]PtdEt accumulation. These results suggest that PGF2 alpha-induced PLD activation is different from PLC-PKC systems. We reported previously that PGF2 alpha stimulated the release of arachidonic acid. The effects of indomethacin, nordihydroguaiaretic acid (NDGA), and 5,8,11,14-eicosatetraynoic acid (ETYA), inhibitors of arachidonic acid metabolism, on PGF2 alpha-stimulated PtdEt accumulation were examined. Pretreatment with indomethacin enhanced PGF2 alpha-induced PtdEt accumulation. In contrast, pretreatment with NDGA and ETYA inhibited PGF2 alpha-induced PtdEt accumulation. It is suggested that PGF2 alpha-stimulated PLD activation is mediated via lipoxygenase products.
Yamamoto
H
H
Department of Obstetrics and Gynecology, Sapporo Medical University, Japan.
Endo
T
T
Kiya
T
T
Goto
T
T
Sagae
S
S
Ito
E
E
Watanabe
H
H
Kudo
R
R
D015117:5,8,11,14-Eicosatetraynoic Acid / Q000494:pharmacology; D000818:Animals; D016718:Arachidonic Acid / Q000037:antagonists & inhibitors*; D016861:Cyclooxygenase Inhibitors / Q000494:pharmacology; D015237:Dinoprost / Q000494:pharmacology*; D004789:Enzyme Activation / Q000187:drug effects; D005260:Female; D020404:Glycerophospholipids; D007213:Indomethacin / Q000494:pharmacology; D007700:Kinetics; D016859:Lipoxygenase Inhibitors / Q000494:pharmacology; D008184:Luteal Cells / Q000187:drug effects / Q000201:enzymology*; D009637:Masoprocol / Q000494:pharmacology; D010712:Phosphatidic Acids / Q000378:metabolism; D010739:Phospholipase D / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley; D014316:Tritium
D016861:Cyclooxygenase Inhibitors; D020404:Glycerophospholipids; D016859:Lipoxygenase Inhibitors; D010712:Phosphatidic Acids; C051521:phosphatidylethanol; D014316:Tritium; D015117:5,8,11,14-Eicosatetraynoic Acid; D016718:Arachidonic Acid; D009637:Masoprocol; D015237:Dinoprost; D010739:Phospholipase D; D007213:Indomethacin