Fertilization and activation currents in bovine oocytes.
One of the first events that occurs at fertilization is a transient modification of the electrical properties of the oocyte plasma membrane. The whole-cell voltage clamp technique was used to demonstrate an outward ion current and a hyperpolarization of the plasma membrane after fertilization in bovine oocytes. These electrical events, together with measurement of internal calcium concentrations, were also recorded after injection with sperm factor and exposure to parthenogenetic activators, such as Ca(2+) ionophore, ethanol and thapsigargin. Experiments were carried out simultaneously in immature and in vitro matured oocytes. Significant differences were recorded in the activation current and hyperpolarization among oocyte activators and between immature and matured oocytes. However, outward ion current and Ca(2+) release showed similar dynamics. The injection of the calcium chelator EGTA completely abolished both ion current and hyperpolarization, indicating that these electrical events are calcium dependent. Addition of specific calcium releasers, such as 1,4,5-inositol trisphosphate (IP(3)) and caffeine, triggered ion activation current and hyperpolarization indicating that IP(3) and ryanodine receptors are active in both immature and matured oocytes. Different ion channel inhibitors were used to characterize the channels underlying outward currents. Only addition of rIberiotoxin caused a complete inhibition of the current, indicating the involvement of high conductance Ca(2+)-activated K(+) channels in generating activation current. In conclusion, these findings provide evidence that bovine oocyte activation is associated with Ca(2+)-dependent electrical events. Oocytes have the potential to react to different activators even when immature; however, oocyte maturation seems to increase sensitivity to physiological activators, such as spermatozoa and sperm factor, and chemicals, such as ethanol.
Tosti
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Cell Biology Unit, Stazione Zoologica Anton Dohrn, Villa Comunale, 80121 Naples, Italy.
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Cuomo
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D000818:Animals; D002110:Caffeine / Q000494:pharmacology; D002118:Calcium / Q000378:metabolism*; D002417:Cattle; D002462:Cell Membrane / Q000502:physiology; D002614:Chelating Agents / Q000494:pharmacology; D004533:Egtazic Acid / Q000494:pharmacology; D000431:Ethanol / Q000494:pharmacology; D005260:Female; D005307:Fertilization in Vitro; D015544:Inositol 1,4,5-Trisphosphate / Q000494:pharmacology; D007476:Ionophores / Q000494:pharmacology; D009865:Oocytes / Q000187:drug effects / Q000378:metabolism*; D009866:Oogenesis; D010312:Parthenogenesis; D018408:Patch-Clamp Techniques; D015221:Potassium Channels / Q000187:drug effects / Q000502:physiology*; D024681:Potassium Channels, Calcium-Activated / Q000037:antagonists & inhibitors; D013084:Sperm-Ovum Interactions / Q000502:physiology*; D013268:Stimulation, Chemical; D019284:Thapsigargin / Q000494:pharmacology
D002614:Chelating Agents; D007476:Ionophores; D015221:Potassium Channels; D024681:Potassium Channels, Calcium-Activated; D002110:Caffeine; D000431:Ethanol; D004533:Egtazic Acid; D019284:Thapsigargin; D015544:Inositol 1,4,5-Trisphosphate; D002118:Calcium