J Vet Sci.  2011 Jun;12(2):133-142. 10.4142/jvs.2011.12.2.133.

Dynamic analysis of Ca2+ level during bovine oocytes maturation and early embryonic development

Affiliations
  • 1College of Veterinary Medicine, Northwest Agriculture and Forestry University, Yangling 712100, China.
  • 2Laboratory of Genetic Resources, Institute of Animal Sciences, Chinese Academy of Agricultural Sciences, Beijing 100193, China. weijunguan301@gmail.com, Yuehui_Ma@hotmail.com

Abstract

Mammalian oocyte maturation and early embryo development processes are Ca(2+)-dependent. In this study, we used confocal microscopy to investigate the distribution pattern of Ca2+ and its dynamic changes in the processes of bovine oocytes maturation, in vitro fertilization (IVF), parthenogenetic activation (PA) and somatic cell nuclear transfer (SCNT) embryo development. During the germinal vesicle (GV) and GV breakdown stage, Ca2+ was distributed in the cortical ooplasm and throughout the oocytes from the MI to MII stage. In IVF embryos, Ca2+ was distributed in the cortical ooplasm before the formation of the pronucleus. In 4-8 cell embryos and morulas, Ca2+ was present throughout the blastomere. In PA embryos, Ca2+ was distributed throughout the blastomere at 48 h, similar to in the 4-cell and 8-cell phase and the morula. At 6 h after activation, there was almost no distribution of Ca2+ in the SCNT embryos. However, Ca2+ was distributed in the donor nucleus at 10 h and it was distributed throughout the blastomere in the 2-8 cell embryos. In this study, Ca2+ showed significant fluctuations with regularity of IVF and SCNT groups, but PA did not. Systematic investigation of the Ca2+ location and distribution changes during oocyte maturation and early embryo development processes should facilitate a better understanding of the mechanisms involved in oocyte maturation, reconstructed embryo activation and development, ultimately improving the reconstructed embryo development rate.

Keyword

bovine; concentration of Ca2+; distribution of Ca2+; embryos; oocytes

MeSH Terms

Aniline Compounds/chemistry
Animals
Calcium/*physiology
Cattle/*physiology
Embryonic Development/*physiology
Female
Fertilization in Vitro/*veterinary
Microscopy, Confocal/veterinary
Oocytes/*physiology
Parthenogenesis/*physiology
Xanthenes/chemistry

Figure

  • Fig. 1 Ca2+ distribution in bovine oocytes during different phases of in vitro maturation. A: Germinal vesicle (GV) stage; B and C: Germinal vesicle breakdown (GVBD) stage; D: MI stage; E: End of MI stage; F: MII stage.

  • Fig. 2 Comparison of Ca2+ fluorescence intensity in bovine oocytes during different phases of in vitro maturation. p < 0.05 (*) and p < 0.01 (**) indicate significant difference from other groups.

  • Fig. 3 Ca2+ distribution in bovine in vitro fertilization (IVF) embryos at different times. A: note the round fluorescent zone displayed in the sperm head.

  • Fig. 4 Comparison of Ca2+ fluorescence intensity in bovine IVF embryos at different times. p < 0.05 (*) and p < 0.01 (**) indicate significant difference from other groups.

  • Fig. 5 Ca2+ distribution in bovine parthenogenetic activation (PA) embryos at different times.

  • Fig. 6 Comparison of Ca2+ fluorescence intensity in bovine parthenogenetic activation embryo at different times. p < 0.05 (*) indicates significant difference from other groups.

  • Fig. 7 Ca2+ distribution in bovine somatic cell nuclear transfer (SCNT) embryos at different times.

  • Fig. 8 Comparison of Ca2+ fluorescence intensity in bovine SCNT embryos at different times. p < 0.05 (*) and p < 0.01 (**) indicate significant difference from other groups.

  • Fig. 9 Comparison of Ca2+ fluorescence intensity in bovine IVF, PA and SCNT embryos at different times. p < 0.05 (*) and p < 0.01 (**) indicate significant difference from other groups.


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