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Int. J. Mol. Sci. 2014, 15(12), 23766-23791; doi:10.3390/ijms151223766

When Isolated at Full Receptivity, in Vitro Fertilized Wheat (Triticum aestivum, L.) Egg Cells Reveal [Ca2+]cyt Oscillation of Intracellular Origin

1
Department of Plant Production and Plant Protection, Institute of Plant Science, Faculty of Agricultural and Environmental Sciences, Kaposvár University, Kaposvár H-7400, Hungary
2
Dipartimento di Scienze Ambientali "G. Sarfatti", University of Siena, Siena 53100, Italy
3
Institute of Physiology, Biochemistry and Animal Health, Faculty of Agricultural and Environmental Sciences, Kaposvár University, Kaposvár H-7400, Hungary
4
Department of Technology of Animal Breeding and Management, Faculty of Agricultural and Environmental Sciences Kaposvár University, Kaposvár H-7400, Hungary
5
Department of Plant Cell Biology, Agricultural Institute, Centre for Agricultural Research, Hungarian Academy of Sciences, Martonvàsàr H-2462, Hungary
*
Author to whom correspondence should be addressed.
Received: 16 April 2014 / Revised: 19 August 2014 / Accepted: 25 September 2014 / Published: 19 December 2014
(This article belongs to the Special Issue Cellular and Molecular Mechanisms of Sperm-Egg Interaction)
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Abstract

During in vitro fertilization of wheat (Triticum aestivum, L.) in egg cells isolated at various developmental stages, changes in cytosolic free calcium ([Ca2+]cyt) were observed. The dynamics of [Ca2+]cyt elevation varied, reflecting the difference in the developmental stage of the eggs used. [Ca2+]cyt oscillation was exclusively observed in fertile, mature egg cells fused with the sperm cell. To determine how [Ca2+]cyt oscillation in mature egg cells is generated, egg cells were incubated in thapsigargin, which proved to be a specific inhibitor of the endoplasmic reticulum (ER) Ca2+-ATPase in wheat egg cells. In unfertilized egg cells, the addition of thapsigargin caused an abrupt transient increase in [Ca2+]cyt in the absence of extracellular Ca2+, suggesting that an influx pathway for Ca2+ is activated by thapsigargin. The [Ca2+]cyt oscillation seemed to require the filling of an intracellular calcium store for the onset of which, calcium influx through the plasma membrane appeared essential. This was demonstrated by omitting extracellular calcium from (or adding GdCl3 to) the fusion medium, which prevented [Ca2+]cyt oscillation in mature egg cells fused with the sperm. Combined, these data permit the hypothesis that the first sperm-induced transient increase in [Ca2+]cyt depletes an intracellular Ca2+ store, triggering an increase in plasma membrane Ca2+ permeability, and this enhanced Ca2+ influx results in [Ca2+]cyt oscillation. View Full-Text
Keywords: in vitro fertilization; wheat (Triticum aestivum, L.) egg cell; cytosolic calcium; egg activation; thapsigargin; intracellular Ca2+ store; endoplasmic reticulum in vitro fertilization; wheat (Triticum aestivum, L.) egg cell; cytosolic calcium; egg activation; thapsigargin; intracellular Ca2+ store; endoplasmic reticulum
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Pónya, Z.; Corsi, I.; Hoffmann, R.; Kovács, M.; Dobosy, A.; Kovács, A.Z.; Cresti, M.; Barnabás, B. When Isolated at Full Receptivity, in Vitro Fertilized Wheat (Triticum aestivum, L.) Egg Cells Reveal [Ca2+]cyt Oscillation of Intracellular Origin. Int. J. Mol. Sci. 2014, 15, 23766-23791.

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