The Effect Of Liangjing To Pig Gamete And Embryonic Development

Artificial insemination technology can reduce the cost of production pig breeding industry, thereby enhancing economic efficiency. And it also can accelerate variety improvement, through the promotion of good quality boar semen. The level of semen preservation is an important part relates to the result of artificial insemination. At present, the widely used method of the preservation of boar semen is room temperature preservation method. In order to improve the utilization efficiency of boars, and promote the development of artificial insemination technology, the dilution and preservation of pig semen will become the most critical technologies.This experiment mainly study the effect on sperm fertilizing capacity and other indicators when added Liang Jing to semen dilution. To determine the influence of Liang Jing on oocytes, first add three different concentrations of Liang Jing(0.5%, 0.75%, and 1%) in oocytes maturation medium M199. After oocytes cultured 1h, they are changed to the normal culture medium. And finally compared the three groups with the blank control group for maturation rates, the results by DAPI staining show that three concentrations of Liang Jing have no adverse effects on porcine oocytes’ maturation. When Liang Jing at different concentrations were put into oocytes maturation culture media, the rates of blastocyst formation of parthenotes were no significant difference, compared with the blank control group. And that result means three concentrations of Liang Jing would not affect the oocytes parthenogenetic embryonic development capacity. Followed by the blastocyst TUNEL apoptosis staining, that results show that the total cell number of blastocysts and the number of apoptotic cells was also no significant difference among the four groups, further instructions that Liang Jing will not affect oocytes subsequent embryo development when adding Liang Jing during oocytes maturation.Embryos were cultured in PZM-3 which contained the same three concentrations Liang Jing after parthenogenetic, and all embryos were turn to normal embryos cultured medium after 4h. Compared with the control group, there were no significant differences in cleavage rate, blastocyst rate and the number of apoptotic cells and the total cell blastocyst among the four groups, suggests that co-cultured with this three concentrations of Liang Jing during embryonic development, the ability of embryo development will not be affected.Put the same three concentrations of Liang Jing into semen dilution, and then used computer assisted sperm analysis system(CASA) to detected sperm which diluted by these semen extender, such as sperm motility, A + B grade level spermatozoa, linear motion sperm motility, sperm average linear velocity(VSL), average sperm side of swing(ALH), sperm motility linearity(LIN), sperm motility swing sex(WOB), sperm motility before tropism(STR) and other indicators. From all consequences, it can be found that the linear motion sperm motility of 0.75% group was significantly higher than 1% group when 30 h, but it was no significant difference compared with the other two groups. When 42 h, the sperm motility of 0.75% group was significantly higher than the control group, and there was no significant difference compared 0.75% group with the other two groups, and the A + B grade level spermatozoa of 0.75% group was significantly higher than the control group and 1% group, and the linear motion sperm motility of 0.75% group was significantly higher than the other three groups. At the same time, through the line chart of sperm indexes which changed with time, it can be found that the change ranges of sperm indexes of 0.75% group were smaller than the change ranges of corresponding indexes of the other three groups, such as the proportion of A-grade and B-grade sperm, sperm motility linear motion, sperm average linear velocity(VSL), average sperm side of swing(ALH), sperm motility linearity(LIN), sperm motility before tropism(STR). Taken together these results can be found that the effects of semen extender contained 0.75% Liang Jing is better.Use these semen which diluted by containing 0.5%, 0.75% and 1% three concentrations Liang Jing semen extender and normal semen in vitro fertilization. The results show that both the cleavage rates, blastocyst rates and the total blastocyst cell number, the number of apoptotic cells were not significantly different among the four groups. These results and data of sperm suggest that the semen extender, containing 0.75% Liang Jing, can maintain sperm motility and viability, but it did not improve the effect of IVF. And further research is needed to the impact of Liang Jing on artificial insemination techniques.