Ca2+ movements between intracellular stores, the cytoplasm and external solution were analysed in murine peritoneal macrophages stimulated by various agonists. The Ca2+ content of intracellular stores was estimated from the amplitude of Ca2+-transients elicited by ionomycin applied in Ca2+-free solution. Both uridine 5′-triphosphate (UTP) and platelet-activating factor (PAF) triggered the release of Ca2+ followed by a sustained influx, during which intracellular stores remained totally empty. In contrast, in the continuous presence of adenosine 5′-triphosphate (ATP), Ca2+ was initially released and then rapidly sequestered again by the stores. ATP-induced store refilling was not related to cell depolarization or to an increase in the intracellular Na+ concentration (two specific consequences of ATP stimulation which are not induced by PAF and UTP). Store refilling was not caused by a signal that ATP would fail to induce (e.g. as a result of receptor desensitization), but was positively controlled by ATP, even in the simultaneous presence of a concentration of PAF which, on its own, would have caused a persistent store depletion. The hypothesis that the signal delivered by ATP involves the sequential activation of phospholipase D and protein kinase C is consistent with the present pharmacological evidence. However, although we found conditions in which Ca2+ stores did not refill in the presence of ATP, this maintained store depletion was not accompanied by a sustained Ca2+ response similar to that elicited by PAF or UTP, suggesting that store depletion is a condition which is necessary, but not sufficient, for inducing Ca2+ influx.
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Alonso-Torre SR, Trautmann A (1993) Calcium responses elicited by nucleotides in macrophages. Interaction between two receptor subtypes. J Biol Chem 268:18640–18647
Alvarez J, Montero M, Garcia-Sancho J (1991) Cytochrome P-450 may link intracellular Ca2+ stores with plasma membrane Ca2+ influx. Biochem J 274:193–197
Bean BP, Friel DD (1990) ATP-activated channels in excitable cells In: Narahashi T (ed) Ion channels, Plenum, New York, pp. 169–203
Billah MM (1993) Phospholipase D and cell signaling. Curr Biol 5:114–123
Billah MM, Eckel S, Mullmann TJ, Egan RW, Siegel MI (1989) Phosphatidylcholine hydrolysis by phospholipase D determines phosphatidate and diglyceride levels in chemotactic peptide-stimulated human neutrophils. J Biol Chem 264:17069–17077
Bruns RF, Miller FD, Merriman RL, Howbert JJ, Heath WF, Kobayaashi E, Takahashi I, Tamaoki T, Nakaao H (1991) Inhibition of protein kinase C by calphostin C is light dependent. Biochem Biophys Res Commun 176:288–293
Donnadieu E, Bismuth G, Trautmann A (1992) Calcium fluxes in T lymphocytes. J Biol Chem 267:25864–25872
Dubyak GR, El-Moatassim C (1993) Signal transduction via P2-purinergic receptors for extracellular ATP and other nucleotides. Am J Physiol 265:C577-C606
Dubyak GR, Cowen DS, Meuller LM (1988) Activation of inositol phospholipid breakdown in HL60 cells by P2-purinergic receptors for extracellular ATP. J Biol Chem 263:18108–18117
El-Moatassim C, Dubyak GR (1992) A novel pathway for the activation of phospholipase D by P2z purinergic receptors in BAC1.2F5 macrophages. J Biol Chem 267:23664–23673
Gordon JL (1986) Extracellular ATP: effects, source and fate. Biochem 1233:309–319
Greenberg S, Di Virgilio F, Steinberg TH, Silverstein SC (1988) Extracellular nucleotide mediate Ca2+ fluxes in J774 macrophages by two distinct mechanisms. J Biol Chem 263:10337–10343
Gruchulla RS, Dinh TT, Kennerly DA (1990) An indirect pathway of receptor-mediated 1,2-diacylglycerol formation in mast cells: IgE receptor-mediated activation of phospholipase D. JImmunol 144:2334–2342
Jamal Z, Martin A, Gomez-Munoz A, Brindley DN (1991) Plasma membrane fractions from rat liver contain a phosphatidate phosphohydrolase distinct from that in the endoplasmic reticulum and cytosol. J Biol Chem 266:2988–2996
Montero M, Alvarez J, Garcia-Sancho J (1992) Control of plasma membrane Ca2+ entry by the intracellular Ca2+ stores. Biochem 1288:519–525
Neher E, Augustine GJ (1992) Calcium gradients and buffers in bovine chromaffin cells. J Physiol (Lond) 450:273–302
Nishizuka Y (1992) Intracellular signaling by hydrolysis of phospholipids and activation of protein kinase C. Science 258:607–614
O'Flaherty JT, Jacobson DP, Redman JF (1988) Bidirectional effets of protein kinase C activators. J Biol Chem 264:6836–6843
Penner R, Fasolato C, Hoth R (1993) Calcium influx and its control by calcium release. Curr Biol 3:368–374
Randriamampita C, Trautmann A (1987) Ionic channels in murine macrophages. I Cell Biol 105:761–769
Randriamampita C, Tsien RY (1993) Emptying of intracellular Ca2+ stores releases a novel small messenger that stimulates Ca2+ influx. Nature 364:809–814
Schütze S, Berkovic D, Tomsing O, Unger C, Krönke M (1991) Tumor necrosis factor induces rapid production of 1′2′diacylglycerol by a phosphatidylcholine-specific phospholipase C. J Exp Med 174:975–988
Zhang BX, Tortorici G, Xu X, Muallem S (1994) Antagonists inactivate the inositol 1,4,5-trisphosphate (Ins-1,4,5-P3)-dependent Ca2+ channel independent of Ins-1,4,5-P3 metabolism. J Biol Chem 269:17132–17135
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Alonso-Torre, S.R., Trautmann, A. Rapid refilling of Ca2+ stores in macrophages stimulated by ATP involves the sequential activation of phospholipase D and protein kinase C. Pflügers Arch. 430, 230–237 (1995). https://doi.org/10.1007/BF00374654
- Calcium stores
- Platelet-activating factor
- Uridine 5′-triphosphate
- Adenosine 5′-triphosphate