Store-operated calcium entry in human oocytes and sensitivity to oxidative stress.
Calcium signaling is a cellular event that plays a key role at many steps of fertilization and early development. However, little is known regarding the contribution of extracellular Ca(2+) influx into the cell to this signaling in gametes and early embryos. To better know the significance of calcium entry on oocyte physiology, we have evaluated the mechanism of store-operated calcium entry (SOCE) in human metaphase II (MII) oocytes and its sensitivity to oxidative stress, one of the major factors implicated in the outcome of in vitro fertilization (IVF) techniques. We show that depletion of intracellular Ca(2+) stores through inhibition of sarco(endo)plasmic Ca(2+)-ATPase with thapsigargin triggers Ca(2+) entry in resting human oocytes. Ba(2+) and Mn(2+) influx was also stimulated following inhibition, and Ca(2+) entry was sensitive to pharmacological inhibition because the SOCE blocker 2-aminoethoxydiphenylborate (2-APB) reduced calcium and barium entry. These results support the conclusion that there is a plasma membrane mechanism responsible for the capacitative divalent cation entry in human oocytes. Moreover, the Ca(2+) entry mechanism described in MII oocytes was found to be highly sensitive to oxidative stress. Hydrogen peroxide, at micromolar concentrations that could mimic culture conditions in IVF, elicited an increase of [Ca(2+)](i) that was dependent on the presence of extracellular Ca(2+). This rise was preventable by 2-APB, indicating that it was mainly due to the enhanced influx through store-operated calcium channels. In sum, our results demonstrate the occurrence of SOCE in human MII oocytes and the modification of this pathway due to oxidative stress, with possible consequences in IVF.
Martín-Romero
Francisco Javier
FJ
Departamento de Bioquímicay Biología Molecular, Reproduction and Development Group (REDES), Universidad de Extremadura, Badajoz-06071, Spain. fjmartin@unex.es
Ortíz-de-Galisteo
Jose Ramón
JR
Lara-Laranjeira
Javier
J
Domínguez-Arroyo
Jose Antonio
JA
González-Carrera
Ernesto
E
Alvarez
Ignacio S
IS
D001896:Boron Compounds / Q000494:pharmacology; D002118:Calcium / Q000378:metabolism*; D020013:Calcium Signaling / Q000187:drug effects; D002462:Cell Membrane / Q000378:metabolism*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005307:Fertilization in Vitro; D006801:Humans; D006861:Hydrogen Peroxide / Q000633:toxicity; D009865:Oocytes / Q000187:drug effects / Q000378:metabolism*; D018384:Oxidative Stress; D053498:Sarcoplasmic Reticulum Calcium-Transporting ATPases / Q000037:antagonists & inhibitors; D019284:Thapsigargin / Q000494:pharmacology
D001896:Boron Compounds; D004791:Enzyme Inhibitors; D019284:Thapsigargin; D006861:Hydrogen Peroxide; C109986:2-aminoethoxydiphenyl borate; D053498:Sarcoplasmic Reticulum Calcium-Transporting ATPases; D002118:Calcium