The Effects Of Heavy Ion Radiation On Sheep Oocytes In Vitro Maturation, Fertilization And Development

In vitro fertilization (IVF) is a process by which egg cells are fertilised by sperm outside the body. It is regarded as the third revolution after artificial insemination (AI) technology, embryo transfer (ET) technology in the field of animal breeding. In recent years, as the development of market economy and the industrialization of sheep cultivation, in vitro fertilization technology has been paid more and more attention. However, because of various factors, the sheep IVF remains unstable and it is difficlut to repeat, the ratio of IVF fertilization and embryo development is low, and the quality of embryo development is poor .Thus, we studied the effect of heavy ion irradiation in vitro maturation of sheep oocytes, in vitro fertilization and early embryonic development, the main results were as follows:1. In maturation medium, adding different ratio concentrations of FSH and LH had significantly affects in vitro maturation of oocytes, when adding 10μg/ml FSH +10μg / ml LH (1:1) to medium the ratio of in vitro maturation of oocytes was 61.2% and the highest.2. Having different ratios of IVM of oocytes after adding different concentrations of sodium pyruvate in maturation medium. Adding 0.2g/L sodium pyruvate ,the ratio of IVM was highest, and adding 0.4,0.6g/L, the ratio of IVM remarkably decreased.3. Having a noticeable effect on in vitro maturation of sheep oocytes after heavy ion irradiation, 1.0 and 1.5Gy could significantly promote the in vitro maturation of oocytes compared with the control group, and the stimulative effects of 1.0Gy was more notable than 1.5Gy, but 2.0Gy irradiation would inhibited the IVM of oocytes.4. During in vitro fertilization of sheep, using frozen or fresh semen, there was no significant difference during the early embryonic development. Therefore, in vitro fertilization experiment, frozen semen could also achieve the desired test purposes.5. Early embryo development has changed significantly after sheep sperm was radiated by heavy ion and then used in vitro fertilization. The irradiation with 1.0 and 1.5Gy could significantly improve cleavage ratio and morula ratio of sheep early embryos compared with the unirradiated control; 1.5 Gy irradiation could significantly improve cleavage ratio, while the morula ratio and blastocyst ratio were markedly reduced.The ratio of cleavage,morula,and blastocysts were remarkably reduced by the irradiation with 2.0Gy.6. The irradiation on sheep oocytes with 0.5,1.0Gy could significantly improve cleavage,morula and blastocyst ratios after in vitro fertilization. And 2.0Gy could remarkably reduce the ratios of cleavage,morula and blastocysts which blastocysts ratio was 0.