Abstract
Alcohols induce mating-structure activation in Chlamydomonas eugametos gametes. From the effect of ethanol on the 32P-labelling of polyphosphoinositides, we conclude that the synthesis of these lipids is stimulated. Biologically inactive concentrations of ethanol (<6%) had no effect on synthesis, but 6–8% ethanol stimulated synthesis for upto 60 min. The 32P incorporated into polyphosphoinositides and phosphatidic acid during ethanol treatment was readily chased out when 1 mM unlabelled Na3PO4 was added. Using a binding assay for inositol 1,4,5-trisphosphate, we show that the production of this phospholipid constituent is dramatically increased after ethanol treatment. This effect, coupled to a rise in intracellular calcium concentration, could explain gamete activation. The significance of these results in explaining other ethanol-induced phenomena in algae is discussed.
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Abbreviations
- Ins(1,4,5)P3 :
-
inositol 1,4,5-trisphosphate
- PtdA:
-
phosphatidic acid
- PtdIns:
-
phosphatidylinositol
- PtdIns(4)P:
-
phosphatidylinositol 4-phosphate
- PtdIns(4,5)P2 :
-
phosphatidylinositol 4,5-bisphosphate
References
Aducci, P., Marra, M. (1990) IP3 levels and their modulation by fusicoccin measured by a novel [3H] IP3 binding assay. Biochem. Biophys. Res. Commun. 168, 1041–1046
Berridge, M.J., Irvine, R.F. (1989) Inositol phosphates and cell signalling. Nature 341, 197–205
Bloodgood, R.A., Levin, E.N. (1983) Transient increase in calcium efflux accompanies fertilization. J. Cell Biol. 96, 397–404
Brederoo, J., de Wildt, P., Popp-Snijders, C., Irvine, R.F., Musgrave, A., van den Ende, H. (1991) Polyphosphoinositol lipids in Chlamydomonas eugametos gametes. Planta 184, 175–181
Cheek, T.R. (1991) Calcium regulation and homeostasis. Curr. Opin. Cell Biol. 3, 199–205
Claes, H. (1980) Calcium ionophore-induced stimulation of secretory activity in Chlamydomonas reinhardtii. Arch. Microbiol. 124, 81–86
Cote, G.G., DePass, A.L., Quarmby, L.M., Tate, B.F., Morse, M.J., Satter, R., Crain, R.L. (1989) Separation and characterization of inositol phospholipids from the pulvini of Samanea saman. Plant Physiol. 90, 1422–1428
Dawson, R.M.C., Eichberg, J. (1965) Diphosphoinositide and triphosphoinositide in animal tissue. Extraction, estimation and changes post mortem. Biochem. J. 96, 634–643
Ferguson, J.E., Hanley, M.R. (1991) The role of phospholipases and phospholipid-derived signals in cell activation. Curr. Opin. Cell Biol. 3, 206–212
Hoek, J.B., Rubin, E. (1990) Alcohol and membrane-associated signal transduction. Alcohol Alcoholism 25, 143–156
Hoek, J.B., Thomas, A.P., Rubin, R., Rubin, E. (1987) Ethanolinduced mobilization of calcium by activation of phosphoinositide-specific phospholipase C in intact hepatocytes. J. Biol. Chem. 262, 682–691
Irvine, R.F., Letcher, A.J., Lander, D.J., Drobak, B.K., Dawson, A.P., Musgrave, A. (1989) Phosphatidylinositol (4,5) bisphosphate and phosphatidyl (4) phosphate in plant tissue. Plant Physiol. 89, 888–892
Irving, G.C.J., Cosgrove, D.J. (1972) Inositol phosphate phosphatases of microbiol origin: the inositol pentaphosphate products of Aspergillus ficuum phytases. J. Bacteriol. 112, 434–438
Kaska, D.D., Piscopo, I.C., Gibor, A. (1985) Intracellular calcium redistribution during mating in Chlamydomonas reinhardtii. Exp. Cell Res. 160, 371–379
Kooijman, R., de Wildt, P., van den Briel, W., Tan, S., Musgrave, A., van den Ende, H. (1990) Cyclic AMP is one of the intracellular signals during the mating of Chlamydomonas eugametos. Planta 181, 529–537
Kriz, R., Linn, L.-L., Sultzman, L., Ellis, C., Heldin, C.-H., Pawson, T., Kopf, J. (1990) Phospholipase C isozymes: structural and functional similarities. Ciba Found. Symp. 150, 112–127
Lehle, L. (1990) Phosphatidyl inositol metabolism and its role in signal transduction in growing plants. Plant Mol. Biol. 15, 647–658
Lewin, R.A., Lee, T.-H., Fang, L.-S. (1982) Effects of various agents on flagellar activity, flagellar autotomy and cell viability in four species of Chlamydomonas (Chlorophyta: Volvocales). Symp. Soc. Exp. Biol. 35, 421–437
Majerus, P.W., Ross, T.S., Cunningham, T.W., Caldwell, K.K., Jefferson, A.B., Bansal, V.S. (1990) Recent insights into phosphatidylinositol signaling. Cell 63, 459–465
Margolis, B., Zilberstein, A., Franks, C., Felder, S., Kremer, S., Ullrich, A., Rhee, S.G., Skorecki, K., Schlessinger, J. (1990) Effect of phospholipase C-gamma overexpression on PDGF induced second messengers and mitogenesis. Science 248, 607–610
Martindale, V.E., Salisbury, J.L. (1990) Phosphorylation of algal centrin is rapidly responsive to changes in the external milieu. J. Cell Sci. 96, 395–402.
Nibbering, P.H., Zomerdijk, T.P.L., van Haastert, P.J.M., van Furth, R. (1990) A competition binding assay for the determination of the inositol (1,4,5)-trisphosphate content of human leucocytes. Biochem. Biophys. Res. Comm. 170, 755–762
Pasquale, S., Goodenough, U.W. (1987) Cyclic AMP functions as a primary sexual signal in gametes of Chlamydomonas reinhardtii. J. Cell Biol. 105, 2279–2292
Peterson, O.H., Wakui, M. (1990) Oscillating intracellular Ca2+ signals evoked by activation of receptors linked to inositol lipid hydrolysis: mechanism of generation. J. Membr. Biol. 118, 93–105.
Pijst, H.L., van Driel, R., Janssens, P.M.W., Musgrave, A., van den Ende, H. (1984) Cyclic AMP is involved in sexual reproduction of Chlamydomonas eugametos FEBS Lett. 174, 132–137
Rincón, M., Chen, Q., Boss, W.F. (1989) Characterization of inositol phosphates in carrot (Daucus carota L.) cells. Plant Physiol. 89, 126–132
Sanders, M.A., Salisbury, J.L. (1989) Centrin-mediated microtubule severing during flagellar excission in Chlamydomonas reinhardtii. J. Cell Biol. 108, 1751–1760
Schuring, F., Smeenk, J.W., Homan, W.L., Musgrave, A., van den Ende, H. (1987) Occurrence of O-methylated sugars in surface glycoconjugates on Chlamydomonas eugametos. Planta 170, 322–327
Schuring, F., Brederoo, J., Musgrave, A., van den Ende, H. (1990) Increase in calcium triggers mating structure activation in Chlamydomonas eugametos. FEMS Microbiol. Lett. 71, 237–240
Snell, W.J., Buchanan, M., Clausell, A. (1982) Lidocaine inhibits fertilization in Chlamydomonas: a possible role for calcium in sexual signalling. J. Cell Biol. 94, 607–612
Wiese, L. (1965) On sexual agglutination and mating type substances (gamones) in isogamous heterothallic Chlamydomonads. I. Evidence of the identity of the gamones with the surface components responsible for sexual flagellar contact. J. Phycol. 1, 46–54
Witman, G.B., Carlson, K., Berliner, J., Rosenbaum, J.L. (1972) Chlamydomonas flagella. 1. Isolation and electrophoretic analysis of microtubules, matrix, membranes and mastigonemes. J. Cell Biol. 54, 507–539
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We thank Dr. P. van Haastert (Biochemistry, University of Groningen, The Netherlands) and his colleagues for introducing us to their Ins(1,4,5)P3 assay, and Ben ten Brink (Molecular Cell Biology, University of Amsterdam, The Netherlands) for information about contractile vacuoles. We also thank Bas Nagelkerken, Marcel van der Vaart, Pieter van der Schoor, Gyuri Fenyvesi and Susan Kenter for their help.
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Musgrave, A., Kuin, H., Jongen, M. et al. Ethanol stimulates phospholipid turnover and inositol 1,4,5-trisphosphate production in Chlamydomonas eugametos gametes. Planta 186, 442–449 (1992). https://doi.org/10.1007/BF00195326
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DOI: https://doi.org/10.1007/BF00195326