Summary
Purified rat peritoneal and pleural mast cells preincubated briefly with radioactively labeled fatty acid were treated with A23187, which bypasses primary receptors in stimulating exocytosis. An enhanced incorporation of fatty acid into phosphatidyl choline (PC) that occurred in parallel with histamine release at 24–25°C was observed and was initially proportional to the total amount of histamine discharged. Enhanced PC labeling and histamine secretion were also proportional at temperatures ranging from 17–37°C. Both radioactive linoleic and palmitic acids were incorporated selectively at the β-position of the glycerol backbone of PC. PC labeling by [3H]choline was not detectably different in control and stimulated cells, and phosphatidic acid did not exhibit selectively enhanced β-acylation. Thus, the stimulated labeling in A23187-treated cells may occur secondary to the action of a phospholipase A2 that favors PC as a substrate.
Other peritoneal cell types exhibit a very similar A23187-stimulated selective labeling of PC. Therefore, autoradiography has been used to provide a direct demonstration that in purified preparations, mast cells are the principal cell type engaged in A23187-elicited incorporation of fatty acid into PC. The efficacy of this approach has relied on special procedures devised to obtain significantly different autoradiographic grain densities between control and stimulated preparations that can be attributed to differences in the level of [3H]palmitate-labeled PC.
Preliminary tests using compound 48/80 as a secretory stimulus for mast cells have identified a similar selectively enhanced PC labeling. In either case, however, consideration of possible relationships between PC metabolism and the secretory process are premature since they have not been tested directly.
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Akesson, B. 1970. Initial esterification and conversion of intraportally injected (1-14C) linoleic acid in rat liver.Biochim. Biophys. Acta 218:57–70
Akesson, B., Elovson, J., Arvidson, G. 1970. Initial incorporation into rat liver glycerolipids of intraportally injected (9, 10-3H)palmitic acid.Biochim. Biophys. Acta 218:44–56
Apitz-Castro, R.J., Mas, M.A., Cruz, M.R., Jain, M.K. 1979. Isolation of homogenous phospholipase A2 from human platelets.Biochem. Biophys. Res. Commun. 91:63–71
Bell, R.L., Majerus, P.W. 1980. Thrombin-induced hydrolysis of phosphatidyl inositol in human platelets.J. Biol. Chem. 255:1790–1792
Billah, M.M., Lapetina, E.G. 1982. Rapid decrease of phosphatidyl inositol 4, 5-bisphosphate in thrombin-stimulated platelets.J. Biol. Chem. 257:12705–12708
Billah, M.M., Lapetina, E., Cuatrecasas, P. 1981. Phospholipase A2 activity specific for phosphatidic acid.J. Biol. Chem. 256:5399–5403
Bligh, E.G., Dyer, W.J. 1959. A rapid method of total lipid extraction and purification.Can. J. Biochem. 37:911–917
Brandt, A.E., Lands, W.E.M. 1967. The effect of acyl-group composition on the rate of acyltransferase-catalyzed synthesis of lecithin.Biochim. Biophys. Acta 144:605–612
Caro, L.G., Van Tubergen, R.P. 1962. High resolution autoradiography. I. Methods.J. Cell Biol. 15:173–188
Castagna, M., Takai, Y., Kaibuchi, K., Sano, K., Kikkawa, V., Nishizuka, Y. 1982. Direct activation of calcium-activated phospholipid-dependent protein kinase by tumor-promoting phorbol esters.J. Biol. Chem. 257:7847–7851
Castle, A.M., Castle, J.D. 1981. The purification and partial characterization of phospholipase A2—a secretory protein of rabbit parotid gland.Biochim. Biophys. Acta 666:259–274
Cochrane, D.E., Douglas, W.W. 1974. Calcium-induced extrusion of secretory granules (exocytosis) in mast cells exposed to 48/80 or the ionophore A23187 and X537A.Proc Natl. Acad. Sci. USA 71:408–412
Cockroft, S., Gomperts, B.D. 1979. Evidence for a role of phosphatidyl inositol turnover in stimulus-secretion coupling.Biochem. J. 178:681–687
Crews, F.T., Morita, Y., McGiyney, A., Hirata, F., Siraganian, R.P., Axelrod, J. 1981. Ig-E-Mediated histamine release in rat basophilic leukemia cells: Receptor activation, phospholipid methylation, Ca2+ flux, and release of arachidonic acid.Arch. Biochem. Biophys. 212:561–571
DeHaas, G.H., Postema, N.M., Nieuwenhuizen, W., Deenen, L.L.M. van 1968. Purification and properties of phospholipase A from porcine pancreas.Biochim. Biophys. Acta 159:103–117
Deutsch, J.W., Kelly, R.B. 1981. Lipids of synaptic vesicles: Relevance to the mechanism of membrane fusion.Biochemistry 20:378–385
Ferber, E., Resch, K. 1973. Phospholipid metabolism of stimulated lymphocytes: Activation of acyl coA: Lysolecithin acyltransferases in microsomal membranes.Biochim. Biophys. Acta 296:355
Fisher, K.A. 1976. Analysis of membrane halves: Cholesterol.Proc. Natl. Acad. Sci. USA 73:173–177
Foreman, J.D., Monger, J.L., Gomperts, B.D. 1973. Calcium ionophores and movement of calcium ions following the physiological stimulus to a secretory process.Nature (London) 245:249–251
Franson, R., Waite, M. 1978. Relation between calcium requirement, substrate charge, and rabbit polymorphonuclear leukocyte phospholipase A2 activity.Biochemistry 17:4029–4033
Frei, E., Zahler, D. 1979. Phospholipase A2 from sheep erythrocyte membranes: Ca2+ dependence and localization.Biochim. Biophys. Acta 550:450–463
Groot, P.H.E., Scholte, H.R., Hulsmann, W.C. 1975. Fatty acid activation: Specificity, localization and function.In: Advance in Lipid Research. R. Paolette and D. Kutcheoski, editors. Vol. 14, pp. 75–126. Academic, New York
Helander, H.F., Bloom, G.D. 1974. Quantitative analysis of mast cell structure.J. Microsc. 100:315–321
Hirata, F., Axelrod, J., Crews, F.T. 1979. Concanavalin A stimulates phospholipid methylation and phosphatidylserine decarboxylation in rat mast cells.Proc. Natl. Acad. Sci. USA,76:4813–4816
Hirata, F., Corcoran, B.A., Vankatasubramanian, K., Schiffmann, E., Axelrod, J. 1979. Chemoattractants stimulate degradation of methylated phospholipids and release of arachidonic acid in rabbit leukocytes.Proc. Natl. Acad. Sci. USA 76:2640–2643
Hirata, F., Toyoshima, S., Axelrod, J., Waxdal, M.J. 1980. Phospholipid methylation: A biochemical signal modulating lymphocyte mitogenesis.Proc. Natl. Acad. Sci. USA 77:862–865
Ho, P.C., Orange, R.P. 1978. Indirect evidence of phospholipase A activation in purified mast cells during reverse anaphylactic challenge.Fed. Proc. 37:1667
Hokin, L.E. 1968. Dynamic aspects of phospholipids during protein secretion.Int. Rev. Cytol. 23:187–208
Ishizaka, T., Hirata, F., Ishizaka, K., Axelrod, J. 1980. Stimulation of phospholipid methylation. Ca2+ influx. and histamine release by bridging IgE receptors on rat mast cells.Proc. Natl. Acad. Sci. USA 77:1903–1906
Kalina, M., Pease, D.C. 1977. The preservation of ultrastructure in saturated phosphatidyl cholines by tannic acid model systems and Type II pneumocytes.J. Cell Biol. 74:726–741
Kennerly, D.A., Sullivan, T.J., Parker, C.W. 1979. Activation of phospholipid metabolism during mediator release from stimulated rat mast cells.J. Immunol. 122:152–159
Kennerly, D.A., Sullivan, T.J., Sylwester, P., Parker, C.W. 1979. Diacylglycerol metabolism in mast cells: A potential role in membrane fusion and arachidonic acid release.J. Exp. Med. 150:1039–1044
Lagunoff, D., Wan, H. 1974. Temperature dependence of mast cell histamine secretion.J. Cell. Biol. 61:809–811
Lewis, R.A., Austen, K.F. 1981. Mediation of local homeostasis and inflammation by leukotrienes and other mast cell dependent compounds.Nature (London) 293:103–108
Lewis, R.A., Soter, N.A., Diamond, P.T., Austen, K.F., Oates, J.A., Roberts, L.J. 1982. Prostaglandin D2 generation after activation of rat and human mast cells with antiIgE.J. Immunol. 129:1627–1631
Luft, J.H. 1961. Improvements in epoxy resin embedding methods.J. Biochem. Biophys. Cytol. 9:409–414
Martin, T.W., Lagunoff, D. 1982. Rat mast cell phospholipase A2: activity toward exogenous phosphatidyl serine and inhibition by N-(7-nitro-2, 1, 3-benzoxadiaxol-4-yl) phosphatidyl serine.Biochemistry 21:1254–1260
McGivney, A., Crews, F.T., Hirata, F., Axelrod, J., Siraganian, R.P. 1981. Rat basophilic leukemia cell lines defective in phospholipid methyltransferase enzymes, Ca2+ influx, and histamine release: Reconstitution by hybridization.Proc. Natl. Acad. Sci. USA 78:6176–6180
McGivney, A., Morita, Y., Crews, F.T., Hirata, F., Axelrod, J., Siraganian, R.P. 1981. Phospholipase activation in the IgE-mediated and Ca2+ ionophore A23187-induced release of histamine from rat basophilic leukemia cells.Arch. Biochem. Biophys. 212:572–580
Michell, R.H. 1975. Inositol phospholipids and cell surface receptor function.Biochim. Biophys. Acta 425:81–147
Morrison, D.C., Roser, J.F., Henson, P.M., Cochrane, C.G. 1974. Activation of rat mast cells by low molecular weight stimuli.J. Immunol. 112:573–582
Palade, G.E. 1975. Intracellular aspects of the process of protein synthesis.Science 189:347–358
Pfenninger, K.H., Johnson, M.P. 1981. Nerve growth factor stimulates phospholipid methylation in growing neurites.Proc. Natl. Acad. Sci. USA 78:7797–7800
Richardson, K.C., Jarett, J., Finke, E.H. 1960. Embedding in epoxy resins for ultrathin sectioning in electron microscopy.Stain Technol. 35:313–323
Rittenhouse-Simmons, S. 1979. Production of diglyceride from phosphatidyl inositol in activated human platelets.J. Clin. Invest. 63:580–587
Roberts, L.J., Lewis, R.A., Oates, J.A., Austen, K.F. 1979. Prostaglandin, thromboxane and 12-hydroxy-5,8,10,14-eicosatetraenoic acid production by ionophore-stimulated rat serosal mast cells.Biochim. Biophys. Acta 575:185–192
Roelofsen, B., Zwaal, R.F.A. 1976. The use of phospholipase in the determination of asymmetric phospholipid distribution in membranes.In: Methods in Membrane Biology. K.D. Korn, editor. Vol. 7, pp. 147–177. Plenum, New York
Rohlich, P., Anderson, P., Uvnas, B. 1971. Electron microscope observations on compound 48/80-induced degranulation in rat mast cells: Evidence for sequential exocytosis of storage granules.J. Cell Biol. 51:465–483
Rutten, W.J., DePont, J.H.M., Bonting, S.L., Daeman, F.M. 1975. Lysophospholipids in pig pancreatic zymogen granules in relation to exocytosis.Eur. J. Biochem. 54:259–265
Saffitz, J.E., Gross, R.W., Williamson, J.R., Sobel, B.E. 1981. Autoradiography of phosphatidyl choline.J. Histochem. Cytochem. 29:371–378
Salpeter, M.M., Bachmann, L. 1965. Assessment of technical steps in electron microscope autoradiography.In: Use of Radioautography in Investigation of Protein Synthesis. C.P. LeBond and E. Warren, editors. pp. 23–41. Academic, New York
Schnar, R., Weigel, P., Kuhlenschmidt, M., Lee, Y., Roseman, S. 1978. Adhesion of chicken hepatocytes to polyacrylamide gels derivatized with N-acetyglucosamine.J. Biol. Chem. 253:7940–7951
Schrey, M.P., Rubin, R.P. 1979. Characterization of a calcium-mediated activation of arachidonic acid turnover in adrenal phospholipids by corticotropin.J. Biol. Chem. 254:11234–11241
Shore, P.A., Burkhalter, A., Cohn, V.H. Jr. 1959. A method for fluorometric assay of histamine in tissues.J. Pharmacol. Exp. Ther. 127:182–186
Sokal, R.R., Rohlf, F.J. 1981.In: Biometry. pp. 429–450. W. H. Freeman, San Francisco
Spector, A.A. 1968. The transport and utilization of free fatty acid.Ann. N.Y. Acad. Sci. 149:768–783
Strandberg, K., Sydbom, A., Uvnas, B. 1975. Incorporation of choline, serine, ethanolamine, and inositol into phospholipids of isolated rat mast cells.Acta Physiol. Scand 94:54–62
Strandberg, K., Westerberg, S. 1976. Composition of phospholipids and phospholipid fatty acids in rat mast cells.Molec. Cell Biochem. 11:103–107
Uvnas, B. 1974. The isolation of secretory granules from mast cells.Methods Enzymol. 51:395–402
Uvnas, B., Thon, I. 1961. Evidence for enzymatic histamine release from isolated rat mast cells.Exp. Cell Res. 23:45–57
Van den Bosch, H. 1980. Intracellular phospholipases A.Biochim. Biophys. Acta 604:191–246
Van den Bosch, H. 1974. Phosphoglyceride metabolism.Annu. Rev. Biochem. 43:243–277
Van den Bosch, H., Deenen, L.L.M. van 1965. Chemical structure and biochemical significance of lysolecithins from rat liver.Biochim. Biophys. Acta. 106:326–337
Van Golde, L.M.G., Van den Bergh, S.G. 1977. The liver.In: Lipid Metabolism in Mammals. F. Snyder, editor. pp. 35–149. Plenum, New York
Wells, M., Hanahan, D. 1968. Phospholipase A fromCrotalus adamanteus venom.Methods Enzymol. 14:178–184
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Castle, J.D., Castle, A.M., Ma, A.K. et al. An enhanced incorporation of fatty acid into phosphatidyl choline that parallels histamine discharge in mast cells. J. Membrain Biol. 79, 215–230 (1984). https://doi.org/10.1007/BF01871061
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DOI: https://doi.org/10.1007/BF01871061