The Verification Of The Differetial Genes In Simulated Micrograity And The Role Of Ndufa4 In Mouse Zygote Development

Dang Nannan using the simulated microgravity environment of Rotating Wall Vessel Bioreactor(RWVB)on zygotes pronucleus stage in vitro culture,Single-celled sequencing showed that Tmem59?1600010M07Rik?Lars2? Nts?Ndufa4?Teddm1b and Papolog gene expression significantly in C57.Mouse zygotes under the different gravity in vitro culture,we using after 20 h h CG the male and female pronucleus appearing as fertilization,Zygotes after 20 h h CG culture 9h in vitro compare with in simulated microgravity 9h.Q-PCR detect these genes express in Kunmingmice,The results shows: Tmem59 ? 1600010M07 Rik ? Lars2 and Ndufa4 genes expression significantly in simulated microgravity condition.66.7% of accuracy compare with Single-celled results.In order to elucidate the Ndufa4 function in mouse zygote development,we produced p EGFP-Ndufa4 in vitro,overexpressed Ndufa4 in mouse zygote by microinjection technique.The blastocyst rate showed significantly when overexpressed Ndufa4 gene.We injected Si RNA of Ndufa4 into mouse zygote by microinjection technique.Results showed after the injection of Ndufa4 Si RNA in mouse zygote,the expression of Ndufa4 was decreased significantly and blastocyst rate showed significantly.Q-PCR detect genes associated with apoptosis like Bax?Bak?Bcl-xl?Bcl-2 and cyt C express after injection p EGFP-Ndufa4 and Ndufa4 Si RNA,we found that both Bax and cyt C genes expression significantly after injection.The expression of Bak gene showed significantly after injected p EGFP-Ndufa4.Ndufa4 maybe related to apoptosis.