Prediction of the blastulation onset in one day behind embryos by the expression profile of small non-coding RNAs

DOI: https://doi.org/10.29296/24999490-2020-05-07

Y.S. Drapkina, A.V. Timofeeva, V.V. Chagovets, N.P. Makarova, A.M. Frolova, E.A. Kalinina Kulakov National Medical Research Center of Obstetrics, Gynecology, and Perinatology, Akademika Oparina str., 4, Moscow, 117997, Russian Federation E-mail: [email protected]

Introduction. Successful embryonic development immediately after fertilization depends on the maternal messenger RNAs (mRNAs) destruction and zygotic genome transcription activation during maternal to zygotic transition (MZT). Key regulators of MZP are small non-coding RNAs (sncRNAs) – microRNAs and piwiRNAs. These molecules can be identified in embryo culture medium and used as a marker of embryo quality, blastulation ability, as well as its implantation potential. The aim this study. To analyze blastulation rate in embryos, which are one day behind in development, according to microRNAs and piwiRNAs expression profile in their culture medium. Methods. The RT-PCR method was used for the quantitative analysis of microRNAs and piwiRNAs in 22 samples of embryo culture media obtained on day 4 after fertilization. Embryos included were one day behind in development and were at the 8-cells stage. The samples were divided into three groups according to morphological and functional characteristics of the embryos on day 6: group I – average/excellent blastocysts (1 – 4AA, 1 – 4BB, 1 – 6BB, 1 – 3AA, 1 – 3BB), group II – poor quality blastocysts (3 – 4СС, 4 – 3СС, 1 – 2СС, 1 – 3ВС, and 1 – 4СВ) and group III – degraded embryos (7). Results. The culture media from embryos which degrade or become poor quality blastocysts on day 6 has almost the same level of microRNA let-7i-5p, but microRNA let-7i-5p expression level is statistically significantly higher in culture medium of the embryos which are capable to develop into average/excellent quality blastocyst. In culture media of embryos which develop into average/excellent quality blastocyst, the level of microRNA let-7b-5p and piwiRNA piR020401 is also statistically significantly higher then in culture media of degraded embryos and embryos which developed into bad quality blastocysts. Conclusion. The data obtained confirm the level of microRNA let-7b-5p, microRNA let-7i-5p and piwiRNA piR020401 to show embryo quality and development potential. Assessment of their expression level can be used for diagnostic and prognostic purposes in selecting the best quality embryo.
Keywords: 
ART, culture medium, embryo, microRNAs, piwiRNAs, pregnancy
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