Effect of vitrification on biogenesis pathway and expression of development-related microRNAs in preimplantation mouse e

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Effect of vitrification on biogenesis pathway and expression of development-related microRNAs in preimplantation mouse embryos Elham Azizi . Marefat Ghaffari Novin . Mohammad Naji . Fardin Amidi . Hossein Hosseinirad . Zahra Shams Mofarahe

Received: 6 November 2019 / Revised: 15 April 2020 / Accepted: 28 September 2020 Ó Springer Nature B.V. 2020

Abstract Vitrification of embryos has been known as the most efficient cryopreservation method in assisted reproductive technology clinics. Vitrification of preimplantation embryo might be associated with altered gene expression profile and biochemical changes of vitrified embryos. Stringent regulation of gene expression in early embryonic stages is very critical for normal development. In the present study, we investigated the effect of vitrification on the canonical miRNA biogenesis pathway, and also the expression of developmental related miRNAs, in 8-cell and blastocyst mouse embryos. Although the

expression pattern of the miRNA biogenesis pathway genes differed between 8-cell and blastocyst mouse embryos, vitrification did not affect the expression level of these genes in preimplantation embryos. The expression levels of miR-21 and let-7a were significantly decreased in vitrified 8-cell embryos and fresh blastocysts when compared with fresh 8-cell embryos. The expression of Stat3 was significantly reduced in blastocysts after vitrification. The alteration in the expression pattern of miRNAs, due to their mode of action, can affect broad downstream key developmental signaling pathways. Therefore, the blastocyst stage

E. Azizi  M. Ghaffari Novin  H. Hosseinirad  Z. Shams Mofarahe (&) Department of Biology and Anatomical Sciences, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran e-mail: [email protected]

M. Naji Urology and Nephrology Research Center (UNRC), Shahid Beheshti University of Medical Sciences, Tehran, Iran e-mail: [email protected]

E. Azizi e-mail: [email protected]

F. Amidi Department of Anatomy, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran e-mail: [email protected]

M. Ghaffari Novin e-mail: [email protected] H. Hosseinirad e-mail: [email protected] M. Ghaffari Novin Infertility and Reproductive Health Research Center (IRHRC), Shahid Beheshti University of Medical Sciences, Tehran, Iran

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is the preferred point for embryo vitrification as they are less susceptible to cryo-damages regarding the stability of miRNAs related to the developmental and implantation competence of embryo. Keywords Preimplantation embryo  Vitrification  microRNA expression  microRNA biogenesis Abbreviations ART Assisted reproductive technology IVF In vitro fertilization miRNAs microRNAs DGCR8 DiGeorge Syndrome Critical Region 8 prePrecursor miRNA miRNA XPO5 Exportin-5 AGO Argonaute RISC RNA-induced silencing complex PMSG Pregnant mare serum gonadotropin hCG Human chorionic gonadotropin ES Equilibrium solution EG Ethylene glycol DMSO Di