Defective calcium release during in vitro fertilization of maturing oocytes of LT/Sv mice
Published: 1 September 2008
Karolina Archacka1,2, Anna Ajduk2, Pawel Pomorski3, Katarzyna Szczepanska2, Marek Maleszewski2 and Maria A. Ciemerych1,2,*
1Department of Cytology and 2Department of Embryology, Institute of Zoology, Faculty of Biology, University of Warsaw and 3Department of Cell Biology, The Nencki Institute of Experimental Biology, Warsaw, Poland
Oocytes of LT/Sv mice have anomalous cytoplasmic and nuclear maturation. Here, we show that in contrast to the oocytes of wild-type mice, a significant fraction of LT/Sv oocytes remains arrested at the metaphase of the first meiotic division and is unable to undergo sperm-induced activation when fertilized 15 hours after the resumption of meiosis. We also show that LT/Sv oocytes experimentally induced to resume meiosis and to reach metaphase II are unable to undergo activation in response to sperm penetration. However, the ability for sperm-induced activation developed during prolonged in vitro culture. Both types of LT/Sv oocytes, i.e. metaphase I and those that were experimentally induced to reach metaphase II, underwent activation when they were fertilized 21 hours after germinal vesicle breakdown (GVBD). Thus, the ability of LT/Sv oocytes to become activated by sperm depends on cytoplasmic maturation rather than on nuclear maturation i.e. on the progression of meiotic division. We also show that sperm penetration induces fewer Ca2+ transients in LT/Sv oocytes than in control wild-type oocytes. In addition, we found that the levels of mRNA encoding different isoforms of protein kinase C (alpha, delta and zeta), that are involved in meiotic maturation and signal transduction during fertilization, differed between metaphase I LT/Sv oocytes which cannot be activated by sperm, and those which are able to undergo activation after fertilization. However, no significant differences between these oocytes were found at the level of mRNA encoding IP3 receptors which participate in calcium release during oocyte fertilization.
LT/Sv, meiosis, in vitro fertilization, activation, calcium oscillations