Studies On Porcine Oocyte In Vitro Maturation (IVM) Pathenogenetic Activation And Apoptosis

Three experiments were carried out in this thesis. The effect of Ca2+ concentration in the electro-activation medium on the parthenogenesis of in vitro maturation(IVM) porcine oocytes was studied in expriment 1. Expriment 2 was to study the effect of culture system on the development of porcine pathenogenetic embryos in vitro. The apoptosis of porcine IVM oocytes was observed in expriment 3.The results of expriment 1 showed that the rates of cleavage, blastocyst and the cells numbers of blastocyst did not exist statistical different when the Ca2+ concentration was from 0.0 to 8.0 mmol/L in the electro-activation medium (p>0.05). However, the blastocyst rate was the highest in 1.0mmol/L Ca2+ medium (13.98?.63). The blastocyst rate was reduced with the increasing of Ca2+ concentration in the medium, but the cell numbers of blastocyst were not different. When the Ca2+ concentration in the medium was improved to 6.5mmol/L or higher the blastocyst rate was reduced to less than 5%.The different culture systems were studied for the development of parthenogenetic porcine embryos in vitro. The electro-activated oocytes were cultured respectively in NCSU23, G2.2 or G2.2-aa for 7d. The cleavage rates didn't exist statistical difference among three systems (p>0.05). The blastocyst rate in NCSU23 (14.37?.77) was significant higher than other groups (p<0.01), but the cell numbers of blastocyst were not different among three systems (p>0.05). The electro-activated oocytes were culutured in NCSU23 for 7 days (A), NCSU23 for 72 hours followed by fresh G2.2 for 4 days (B), G2.2 for 72 hours followed by fresh G2.2 for 4 days (C), and G2.2 for 72 hours followed by fresh NCSU23 for 4 days (D). No statistical difference was found in the cleavage rates among the groups (p>0.05), but the rate of group C was highest (61.81?.79). The blastocyst rate in the group A (12.50+1.49) was significant higher than other groups (p<0.05), but the cell numbers of blastocyst were not statistical different (p>0.05).The electro-activated oocytes were culutured in NCSU23, G2.2-aa, G2.2-aa supplemented with 5mmol/L hypotaurine and 0.6mmol/L cysteine, G2.2-aa supplemented with 5mg/L insulins, G2.2-aa supplemented with 5mg/L insulin, 5mmol/L hypotaurine and0.6mmol/L cysteine for 7 days respectively. No statistical difference was found in the cleavage rates (p>0.05) among the groups. The blastocyst rate in NCSU23(12.24?.63) was significantly higher than other groups (p<0.05). The additions of hypotaurine, systeine or insulin in G2.2-aa were beneficial to blastocyst rates of the porcine parthenogenetic embryos, but were no benefits to the cell numbers of blastocyst (p>0.05).The apoptosis of porcine IVM oocytes was observed using morphologically observed and dyeing Hoechst33342. The rates of cytoplast uneqal cleavage were the higest than the necrotic and the cytoplast eqal cleavage in the group of inactivated oocytes. The cytoplast uneqal cleavage was appeared in 24 hours and the highest rate was appeared in 132 hours. And there was the more amplitude among 24~96 hours. The cytoplast eqal cleavage was appeared in 12 hours and the highest rate was appeared in 120 hours. And there was the more amplitude among 24~72 hours. The necrotic was appeared in 12 hours and sustainingly increased. The more amplitude of necrotic was appeared in 24-108 hours. In the group of electro-activated oocytes, the cytoplast eqal cleavage was mostly trend. Rut the rates of necrotic was higher than cytoplast uneqal cleavag after 60 hours. The cytoplast eqal cleavage was appeared in 12 hours and the highest rate was appeared in 60 hours. And there was the more amplitude among 24~60 hours. The cytoplast uneqal cleavage was appeared in 24 hours and sustainingly increased. And there was the more amplitude among 24~72 hours. The necrotic was appeared in 12 hours and the more amplitude of necrotic was appeared in 24~108 hours. In addition, the rates of cytoplast eqal cleavage in the group of electro-activated oocytes were significant higher than the inactivated groups (p<0.05) a...