Phospholipase D activity of human amnion cells stimulated with phorbol ester and bradykinin.
We investigated the activity of phospholipase D (PLD) in human amnion cells labeled with [3H]oleate. The PLD activity was detected as signal-induced synthesis of phosphatidic acid (PA) and in the presence of ethanol, phosphatidylethanol (PEt). The PLD was shown to be activated by phorbol, 12-myristate, 13-acetate (PMA), calcium ionophore A23187, oxytocin, bombesin and bradykinin, but not by platelet-activating factor (PAF) and epidermal growth factor (EGF). The amniotic PLD thus appeared to be activated by a variety of agonists but with a certain specificity to stimulators. We examined the mode of the PLD activation using PMA (20 nM) and bradykinin (1 microM) as model stimulators. PMA and bradykinin elicited a rapid and sustained response with the peaks of PA-labeling attained at 5 and < 1 min after stimulation, respectively. In both cases, there was a concomitant rise of diacylglycerol (DG), and the PA accumulation was suppressed by ethanol at the expense of labeling of PEt. The PA synthesis caused by the two stimulators was similarly inhibited by staurosporine and by a chronic treatment with PMA (100 nM for 24 h), suggesting that the activation of PLD is linked to the action of protein kinase C. With the cells labeled with radioactive choline and ethanolamine, we found that the amniotic PLD hydrolyzed almost equally phosphatidylcholine and phosphatidylethanolamine. Although bradykinin and PMA stimulated cellular PLD to a comparable extent, prostaglandin (PG)E2 release was not stimulated by bradykinin in contrast to the marked effect by PMA. Further work is thus needed to clarify the significance of the novel PLD signaling pathway in the function of amnion cells.
Department of Obstetrics and Gynecology, Sapporo Medical College, Japan.
M
M
Mizunuma
S
S
Tanaka
R
R
Kudo
H
H
Kanoh
D000650:Amnion / Q000378:metabolism*; D001920:Bradykinin / Q000494:pharmacology*; D002478:Cells, Cultured / Q000187:drug effects; D004075:Diglycerides / Q000032:analysis; D004789:Enzyme Activation / Q000187:drug effects; D020404:Glycerophospholipids; D006801:Humans; D019301:Oleic Acid; D009829:Oleic Acids / Q000378:metabolism; D010712:Phosphatidic Acids / Q000032:analysis*; D010739:Phospholipase D / Q000378:metabolism*; D015398:Signal Transduction; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology*; D014316:Tritium
D004075:Diglycerides; D020404:Glycerophospholipids; D009829:Oleic Acids; D010712:Phosphatidic Acids; C051521:phosphatidylethanol; D014316:Tritium; D019301:Oleic Acid; D010739:Phospholipase D; D013755:Tetradecanoylphorbol Acetate; D001920:Bradykinin