Effect of Sperm Cryopreservation on miRNA Expression and Early Embryonic Development

Xiaoyu Xu; Wanqiong Li; Lina Zhang; Yazhong Ji; Jiaying Qin; Lu Wang; Mingwen Wang; Lingbin Qi; Jinfeng Xue; Bo Lv; Xunyi Zhang; Zhigang Xue

doi:10.3389/fcell.2021.749486

Effect of Sperm Cryopreservation on miRNA Expression and Early Embryonic Development

Front Cell Dev Biol. 2021 Dec 22:9:749486. doi: 10.3389/fcell.2021.749486. eCollection 2021.

Authors

Xiaoyu Xu¹, Wanqiong Li², Lina Zhang², Yazhong Ji², Jiaying Qin¹, Lu Wang¹, Mingwen Wang¹, Lingbin Qi¹, Jinfeng Xue¹, Bo Lv¹, Xunyi Zhang², Zhigang Xue^{1

2}

Affiliations

¹ Department of Regenerative Medicine, Tongji University School of Medicine, Shanghai, China.
² Reproductive Medicine Center, Tongji Hospital, Tongji University School of Medicine, Shanghai, China.

Abstract

Although sperm preservation is a common means of personal fertility preservation, its effects on embryonic development potential need further investigation. The purpose of this study was to identify key microRNA (miRNA) in cryopreserved sperm and determine the changes of these miRNAs and their target genes during embryonic development using cryopreserved sperm. Moreover, the embryonic development potential of cryopreserved sperm was estimated in assisted reproductive technology (ART), where key miRNAs and target genes were validated in sperm and subsequent embryos. Clinical data of embryonic development from cryopreserved sperm indicated a significant decrease in fertilization rate in both in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI) cases, as well as a reduction in blastocyst formation rate in ICSI cases. Meanwhile there was a significant increase in blocked embryo ratio of Day1, Day2, and Day3.5 embryos when frozen-thawed mouse sperm was used, compared with fresh mouse sperm, suggesting a potential negative effect of sperm cryopreservation on embryonic development. From frozen-thawed and fresh sperm in humans and mice, respectively, 21 and 95 differentially expressed miRNAs (DEmiRs) were detected. miR-148b-3p were downregulated in both human and mouse frozen-thawed sperm and were also decreased in embryos after fertilization using cryopreserved sperm. Target genes of miR-148b-3p, Pten, was identified in mouse embryos using quantitative real-time PCR (qRT-PCR) and Western blot (WB). In addition, common characters of cryopreservation of mouse oocytes compared with sperm were also detected; downregulation of miR-148b-3p was also confirmed in cryopreserved oocytes. In summary, our study suggested that cryopreservation of sperm could change the expression of miRNAs, especially the miR-148b-3p across humans and mice, and may further affect fertilization and embryo development by increasing the expression of Pten. Moreover, downregulation of miR-148b-3p induced by cryopreservation was conserved in mouse gametes.

Keywords: embryonic development; gamete cryopreservation; miR-148b-3p; miRNA expression; sperm cryopreservation.