Diagnosis of DNA damage on oocytes which are subject to microinjection and piezo electric as well as determining the impact of estrogen receptor gene polymorphisms on fertilization

In this study, comet assay was applied on four groups of metaphase-II oocytes collected from 104 women undergoing in vitro fertilization (IVF) treatment. Group1 oocytes were not microinjected. Group2 oocytes were the unfertilized ones. Group3 oocytes were exposed to piezo electric stimulation. Group4 was the positive control in which the oocytes were incubated in hydrogen peroxide. It was expected to reveal the impact of microinjection and piezo electric stimulation on oocyte DNA. Group1 and 2 oocytes were found to have lower DNA fragmentation than Group3 and 4.Another point was to detect estrogen receptor gene polymorphisms via PCR-RFLP and fragment analysis and correlate with oocyte maturation and fertilization rates. 104 infertile and 107 fertile women were investigated. First the severity of polymorphisms between infertile and fertile women then the effect of the polymorphisms on IVF parameters was analyzed.Significant statistical relationship was observed between infertile and fertile women in terms of c.454-397T>C, c.454-351A>G and TA polymorphisms. Maturation rates, embryo number and fertilization rates were significantly smaller in women having pp and xx genotypes. Long TA repeat was found to be associated with low embryo number and fertilization rate.The outcome of our study implies the importance of comet technique upon determing the impact of techniques applied in IVF treatment on oocyte DNA and developing the applied techniques. The relationship we found between the polymorphisms and infertility&IVF parameters carry weight in developing and applying treatment methods suitable to individuals genotypes, in this way in increasing the success rates in IVF treatments.