Abstract
Chlamydomonas eugametos gametes of opposite mating type make cell-cell contact via their flagellar surfaces. This contact triggers an increase in the intracellular level of cyclic AMP (cAMP) and several cellular responses which are necessary for cell fusion. Here, we show that wheat-germ agglutinin, which binds to the flagellar surface and induces all mating responses, also increased the intracellular cAMP level. Dibutyryl-cAMP added to non-mating gametes induced flagellar twitching, cell-wall lysis, mating-structure activation, flagellartip activation and an increase in agglutinability. It did not induce agglutinin transport to the flagellar tip (tipping) and may not be the direct cause of flagellar twitching and flagellar-tip activation. In non-illuminated cells, dibutyryl-cAMP was far more effective in evoking mating reactions than in illuminated cells. Light induced a 50% decrease in the cAMP level within 1 min. Adenylate cyclase was found to be associated with cell membranes but only 8% of the total was present in the gamete flagella.
Similar content being viewed by others
Abbreviations
- db-cAMP:
-
dibutyryl-cAMP
- FTA:
-
flagellar tip activation
- Mab:
-
monoclonal antibody
- mt −/mt+ :
-
mating-type minus/plus
- WGA:
-
wheat-germ agglutinin
References
Adair, W.S., Monk, B.C., Cohen, R., Goodenough, U.W. (1982) Sexual agglutinins from the Chlamydomonas flagellar membrane. Partial purification and characterization. J. Biol. Chem. 257, 4593–4602
Berridge, M.J., Irvine, R.F. (1989) Inositol phosphates and cell signalling. Nature 341, 197–205
Bessen, M., Fay, R.B., Witman, G.B. (1980) Calcium control of waveform in isolated flagellar axonemes of Chlamydomonas. J. Cell Biol. 86, 446–455
Bloodgood, R.A. (1987) Glycoprotein dynamics in the Chlamydomonas flagellar membrane. Adv. Cell Biol. 1, 97–130
Bloodgood, R.A., Levin, E.N. (1983) Transient increase in calcium efflux accompanies fertilization in Chlamydomonas. J. Cell Biol. 97, 397–404
Bonini, N.M., Nelson, D.L. (1988) Differential regulation of Paramecium ciliary motility by cAMP and cGMP. J. Cell Biol. 106, 1615–1623
Castagna, M., Takai, Y., Kaibuchi, K., Sano, K., Kikkawa, U., Nishizuka, Y. (1982) Direct activation of calcium-activated, phopholipid-dependent protein kinase by tumor promoting phorbol esters. J. Biol. Chem. 257, 7847–7852
Claes, H. (1980) Calcium ionophore-induced stimulation of secretory activity in Chlamydomonas reinhardii. Arch. Microbiol. 124, 81–86
Cohen, R.J., Atkinson, M.M. (1978) Activation of Phycomyces adenosine 3′, 5′-monophosphate phosphodiesterase by blue light. Biochem. Biophys. Res. Comm. 83, 616–621
Collin-Osdoby, P., Adair, W.S. (1985) Characterization of the purified Chlamydomonas minus agglutinin. J. Cell Biol. 101, 1144–1152
Crabbendam, K.J., Nanninga N., Musgrave, A., van den Ende, H. (1984) Flagellar tip activation in vis-à-vis pairs of Chlamydomonas eugametos. Arch. Microbiol. 138, 220–223
Crabbendam, K.J., Klis, F.M., Musgrave, A., van den Ende, H. (1987) Ultrastructure of the plus and the minus sexual agglutinins of Chlamydomonas eugametos, as visualized by negative staining. J. Ultrastruct. Res. 96, 151–159
Demets, R., Tomson, A.M., Stegwee, D., van den Ende, H. (1988) Cell-cell adhesion in conjugating Chlamydomonas gametes: a self-enhancing process. Protoplasma 145, 27–36
Elzenga, J.T.M., Nanninga, N., Lens, P.F., van den Ende, H. (1982) Flagellar tip activation in Chlamydomonas eugametos. FEMS Microbiol. Lett. 15, 279–283
Goodenough, U.W. (1989) Cyclic AMP enhances the sexual agglutinability of Chlamydomonas flagella. J. Cell Biol. 109, 247–252
Homan, W.L., Musgrave, A., Molenaar, E.M., van den Ende, H. (1980) Isolation of monovalent sexual binding components from Chlamydomonas flagellar membranes. Arch. Microbiol. 128, 120–125
Homan, W.L., Gijsberti Hodenpijl, P., Musgrave, A., van den Ende, H. (1982) Reconstitution of biological activity in isoagglutinins from Chlamydomonas eugametos. Planta 155, 529–535
Homan, W.L., Sigon, C.A.M., van den Briel, W., Wagter, R., de Nobel, H., Mesland, D.A.M., Musgrave, A., van den Ende, H. (1987) Transport of membrane receptors and the mechanics of sexual cell fusion in Chlamydomonas eugametos. FEBS Lett. 215, 323–326
Homan, W.L., Musgrave, A., de Nobel, H., Wagter, R., Kolk, A.H.J., de Wit, D., van den Ende, H. (1988) Monoclonal antibodies directed against the sexual binding site of Chlamydomonas eugametos gametes. J. Cell Biol. 107, 177–189
Hyams, J.S., Borisy, G.G. (1978) Isolated flagellar apparatus of Chlamydomonas: characterization of forward swimming and alteration of waveform and reversal of motion by calcium ions in vitro. J. Cell Sci. 33, 235–253
Irvine, R.F., Letcher, A.J., Lander, D.J., Drobak, B.K., Dawson, A.P., Musgrave, A. (1989) Phosphatidylinositol(4,5)bisphosphate and phosphatidylinositol(4)phosphate in plant tissues. Plant Physiol. 89, 888–892
Janssens, P.M.W., van Essen, H.W., Guijt, J.J.M., de Waal, A., van Driel, R. (1987) Cell fractionation, detergent sensitivity and solubilization of Dictyostelium adenylate cyclase and guanylate cyclase. Mol. Cell. Biochem. 76, 55–65
Kalshoven, H.W., Musgrave, A., van den Ende, H. (1990) Mating receptor complex in the flagellar membrane of Chlamydomonas eugametos gametes. Sex. Plant Reprod. 3, 77–87
Kamiya, R., Witman, G.B. (1984) Submicromalar levels of calcium control the balance of beating between the flagella in demembranated models of Chlamydomonas. J. Cell Biol. 98, 97–107
Kaska, D.D., Piscopo, I.C., Gibor, A. (1985) Intra-cellular calcium redistribution during mating in Chlamydomonas reinhardtii. Exp. Cell Res. 160, 371–379
Klis, F.M., Samson, M.R., Touw, E., Musgrave, A., van den Ende, H. (1985) Sexual agglutination in the unicellular green alga Chlamydomonas eugametos. Identification and properties of the mating type plus agglutination factor. Plant Physiol. 79, 740–745
Kooijman, R., Elzenga, T.J.M., de Wildt, P., Musgrave, A., Schuring, F., van den Ende, H. (1986) Light dependence of sexual agglutinability in Chlamydomonas eugametos. Planta 169, 370–378
Kooijman, R., de Wildt, P., Homan, W.L., Musgrave, A., van den Ende, H. (1988) Light affects agglutinability in Chlamydomonas eugametos by modification of the agglutinin molecules. Plant Physiol. 86, 216–223
Kooijman, R., de Wildt, P., Beumer, S., van der Vliet, G., Homan, W.L., Musgrave, A., Crabbendam, K., van den Ende, H. (1989) Wheat germ agglutinin induces mating reactions in Chlamydomonas eugametos by crosslinking agglutinin-associated glycoproteins in the flagellar membrane. J. Cell Biol. 109, 1677–1687
Mesland, D.A.M. (1976) Mating in Chlamydomonas eugametos: A scanning electron microscopical study. Arch. Microbiol. 109, 31–35
Mesland, D.A.M., Hoffman, J.L., Caligor, E., Goodenough, U.W. (1980) Flagellar tip activation stimulated by membrane adhesions in Chlamydomonas gametes. J. Cell Biol. 84, 599–617
Musgrave, A., van den Ende, H. (1987) How Chlamydomonas court their partners. Trends Biochem. Sci. 144, 470–473
Musgrave, A., van Eijk, E., te Welscher, R., Broekman, R., Lens, P.F., Homan, W.L., van den Ende, H. (1981) Sexual agglutination factor from Chlamydomonas eugametos. Planta 153, 362–369
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
Pijst, H.L.A. (1985) Sexual agglutination of the green alga Chlamydomonas eugametos: symtoms and signals. Thesis, University of Amsterdam, The Netherlands
Pijst, H.L.A., Ossendorp, F.A., van Egmond, P., Kamps, A.M.I.E., Musgrave, A., van den Ende, H. (1984) Sex-specific binding and inactivation of agglutination factor in Chlamydomonas eugametos. Planta 160, 529–535
Pijst, H.L.A., 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, 132–136
Saito, T., Tsubo, Y., Matsuda, Y. (1985) Synthesis and turnover of cell body-agglutinin as a pool for flagellar surface-agglutinin in Chlamydomonas reinhardtii. Arch. Microbiol. 146, 25–29
Samson, M.R., Klis, F.M., Crabbendam, K.J., van Egmond, P., van den Ende, H. (1987) Purification, visualisation and characterization of the sexual agglutinins of the green alga Chlamydomonas moewusii yapensis. J. Gen. Microbiol. 133, 3183–3191
Schuring, F., Smeenk, J.W., Homan, W.L., Musgrave, A., van den Ende H. (1987) Occurence of O-methylated sugars in surface glycoconjugates on Chlamydomonas eugametos. Planta 170, 322–327
Snell, W.J., Moore, W.S. (1980) Aggregation-dependent turnover of flagellar adhesion molecules in Chlamydomonas gametes. J. Cell Biol. 84, 203–210
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
Steiner, A.L., Parker, C.W., Kipnis, D.M. (1972) Radioimmunoassay for cyclic nucleotides: I. preparation of antibodies and iodonated cyclic nucleotides. J. Biol. Chem. 247, 1106–1113
Tash, J.S., Kakar, S.S., Means, A.R. (1984) Flagellar mobility requires the cAMP-dependent phosphorylation of a heat-stable NP-40-soluble 56 kD protein, axonin. Cell 38, 551–559
Tomson, A.M., Demets, R. (1989) Sexual communication in the green alga Chlamydomonas eugametos. Ph. D. thesis, University of Amsterdam
Witman, G.B.K., Carlson, J., Berliner, J., Rosenbaum, J.L. (1972) Chlamydomonas flagella. I. Isolation and electrophoretic analysis of microtubules, matrix, membranes and mastigonemes. J. Cell Biol. 54, 507–539
Author information
Authors and Affiliations
Additional information
We gratefully acknowledge the fruitful discussions with Dr. Rainer Gilles of the Department of Biochemistry at the University of Cologne (FRG), and the advice generously given by Dr. Roel van Driel of the Department of Biochemistry at the University of Amsterdam (The Netherlands).
Rights and permissions
About this article
Cite this article
Kooijman, R., de Wildt, P., van den Briel, W. et al. Cyclic AMP is one of the intracellular signals during the mating of Chlamydomonas eugametos . Planta 181, 529–537 (1990). https://doi.org/10.1007/BF00193006
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00193006