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Song HJ, Kang EJ, Kim MJ, Ock SA, Jeon BG, Lee SL, Rho GJ: Influence of Parthenogenetic Activation on Nuclear Maturation of Canine Oocytes. J Vet Med Sci. 2010 Mar 10.
This study was designed to evaluate the nuclear maturation and maturation promoting factor (MPF) level at different maturation time, and the effect of parthenogenetic activation on nuclear maturation in canine oocytes. Cumulus-oocyte complexes (COCs) were matured in TCM-199 supplemented with 10% fetal bovine serum, hormones, 0.57 mM cysteine, and 10 ng/ml epidermal growth factor for 72 hr at 38.5. In Experiment 1, COCs at 0, 24, 48 and 72 hr of culture were assessed for nuclear maturation and MPF levels using histone H1 kinase activity assay. A significantly higher rate of oocytes at 72 hr than 0, 24 and 48 hr of culture developed to metaphase I-anaphase I and metaphase II. Relative abundance of histone H1 kinase activity of oocytes matured for 48 hr increased to ~1.5 x with a dramatic increase to ~2.5 x for 72 hr, significantly higher than others. In Experiment 2, oocytes matured for 48 hr were parthenogenetically activated with 5 mum ionomycin for 5 min (Group 1) and followed by 10 mug/ml of cycloheximide for 3 hr (Group 2), or no treatment (Control). Oocytes were then cultured for 24 hr and assessed for nuclear maturation. A significantly higher rate of oocytes in Group 1 developed to metaphase II than in Group 2 and control. These results indicated that ionomycin treatment at 48 hr of in vitro maturation had a positive influence on the oocyte progression to metaphase II stage. |
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