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Differential contribution of cytoplasmic Ca2+ and Ca2+ influx to gamete fusion and egg activation in maize

Nature Cell Biology volume 3, pages 11201123 (2001) | Download Citation

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Abstract

In multicellular organisms, gamete fusion triggers a set of events, collectively known as egg activation, that leads to the development of a new individual. Every species that has been studied shows at least one rise in cytoplasmic Ca2+ concentration ([Ca2+]Cyt) after gamete fusion1 which is believed to be involved in activation. Yet the source and regulation of this Ca2+ signal and the way it is transduced inside the zygote are controversial1. In higher plants, in vitro fertilization (IVF)2,3 has enabled the description of a rise in [Ca2+]Cyt (ref. 4) that is sufficient for activation5, and of a Ca2+ influx that spreads as a wavefront from the fusion site5 The relationship between these two responses is unknown. Using a new combination of methods that simultaneously monitor the extracellular flux with a Ca2+-vibrating probe, and [Ca2+]Cyt by widefield imaging, we directly determined that the Ca2+ influx precedes the [Ca2+]Cyt elevation by 40–120 s. In addition, results from experiments using the Ca2+-channel inhibitor gadolinium (Gd3+) suggest that the Ca2+ influx may be necessary for sperm incorporation. We also present evidence for a putative sperm-dependent Gd3+-insensitive localized Ca2+ influx confined to the fusion point.

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Acknowledgements

The present collaboration between the ENS and IGC was funded by a ICCTI/CNRS protocol (9607/CNRS). We thank M. Beckert (INRA, Clermont-Ferrand) for providing A188, A6 and DH5×DH7 seeds. We also thank F. Rozier for technical help and C. Arnoult for very helpful comments. J.A.F. and the vibrating-probe set-up were partly funded by Fundação Luso-Americana para o Desenvolvimento and Fundação para a Ciência e Tecnologia (PRAXIS/C/BIA/11034/1998). J.-E.F. was supported by CNRS.

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Affiliations

  1. Laboratoire Reproduction et Développement des Plantes, UMR 5667 CNRS/INRA/ENS/Université Lyon I, École Normale Supérieure de Lyon, 46 allée d'Italie, F-69364 Lyon, France cedex 07

    • Anne-Frédérique Antoine
    • , Jean-Emmanuel Faure
    •  & Christian Dumas
  2. Instituto Gulbenkian de Ciência, PT-2780–156 Oeiras, Portugal

    • José A. Feijó
  3. Centro de Biotecnologia Vegetal, Faculdade de Ciências, Universita de Lisboa, Campo Grande, Ed. C2. PT-1749–016 Lisboa, Portugal

    • José A. Feijó
  4. Present adress: Laboratoire de Biologie du Développement, SN3, Université des Sciences et Technologies de Lille, F-59655 Villeneuve d'Ascq Cedex, France

    • Anne-Frédérique Antoine

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Correspondence to Anne-Frédérique Antoine.

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DOI

https://doi.org/10.1038/ncb1201-1120

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