thapsigargin and safingol

Our previous studies show that epidermal growth factor (EGF) stimulates phospholipase D (PLD)-induced phosphatidic acid (PA) formation in rat calvarial osteoblastic cells. This study investigated the effects of PA on cytosolic calcium ([Ca2+]i) and proliferation, and the possible involvement of the PLD pathway in EGF effects on [Ca2+]i and proliferation in rat calvarial osteoblastic cells. PA markedly increased [Ca2+]i. This response was unaffected by thapsigargin, which depletes [Ca2+]i pools, blocked by verapamil, a calcium channel blocker, and enhanced by propanolol, an inhibitor of PA-phosphohydrolase. PA also reduced the EGF dependent-[Ca2+]i increase by 60%, while a PLD inhibitor blocked these effects. Furthermore, PA significantly increased cell proliferation (P < 0.05) which was inhibited by verapamil and enhanced by H-7 (PKC inhibitor). The PLD inhibitor significantly (P < 0.05) reduced the EGF-induced increase in proliferation. In summary, PA stimulates rat calvarial osteoblastic cell proliferation and mobilization of [Ca2+]i using extracellular pools, and EGF's mitogenic effect on these cells requires activation of PLD.

Topics: 1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine; Animals; Calcium; Calcium Channel Blockers; Calcium Signaling; Cell Division; Cells, Cultured; Cytoplasm; Epidermal Growth Factor; Osteoblasts; Phosphatidic Acids; Phospholipase D; Propranolol; Protein Kinase C; Rats; Rats, Sprague-Dawley; Skull; Sphingosine; Thapsigargin; Verapamil