Calcium release and subsequent development induced by modification of sulfhydryl groups in porcine oocytes

The mechanism of Ca2+ release induced by modification of sulfhydryl groups and the subsequent activation of porcine oocytes were investigated, Thimerosal, a sulfhydryl-oxidizing compound, induced Ca2+ oscillation in matured oocytes, In thimerosal-preincubated oocytes, the amount of Ca2+ released after microinjection of inositol 1,4,5-trisphosphate (InsP(3),) or ryanodine increased strikingly, indicating that thimerosal potentiated both InsP(3)-,- and ryanodine-sensitive Ca2+ release pathways. Thimerosal also enhanced the sensitivity of oocytes to microinjected Ca2+ so that in pretreated oocytes a Ca2+ injection triggered a larger transient. Heparin at concentrations that normally block the InsP(3),-induced Ca2+ release were without effect; higher doses significantly increased the time leading up to the first spike. The thimerosal-induced Ca2+ release could not be blocked by procaine, and it did not require the formation of InsP, since pre-injection with neomycin did not prevent the oscillation, Immunocytochemistry revealed that thimerosal treatment destroyed the meiotic spindle, preventing further development, an effect that could be reversed by dithiothreitol. The combined thimerosal/dithiothreitol treatment triggered second polar body extrusion in 50% of the oocytes, and as a result of this activation scheme similar to 15% of the in vitro- and similar to 60% of the in vivo-matured oocytes developed to blastocyst during a 7-day culture in vitro.