Effect of Sperm Cryopreservation on miRNA Expression and Early Embryonic Development.
Effect of Sperm Cryopreservation on miRNA Expression and Early Embryonic Development.
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精子冷冻保存对 miRNA 表达和早期胚胎发育的影响
DOI：
10.3389/fcell.2021.749486
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发表时间：
2021
影响因子：
5.5
通讯作者：
Xue Z
中科院分区：
文献类型：
--
作者：
Xu X;Li W;Zhang L;Ji Y;Qin J;Wang L;Wang M;Qi L;Xue J;Lv B;Zhang X;Xue Z
虽然精子保存是个人生育能力保存的常用手段，但其对胚胎发育潜力的影响需要进一步研究。本研究的目的是鉴定冷冻精子中的关键microRNA（miRNA），并确定这些miRNA及其靶基因在冷冻精子胚胎发育过程中的变化。此外，在辅助生殖技术（ART）中评估了冷冻保存精子的胚胎发育潜力，其中在精子和随后的胚胎中验证了关键miRNA和靶基因。冷冻保存精子胚胎发育的临床数据表明，体外受精（IVF）和卵胞浆内单精子注射（ICSI）病例的受精率显著降低，ICSI病例的囊胚形成率也降低。同时，与新鲜小鼠精子相比，冷冻小鼠精子在第1、2和3.5天胚胎中的阻滞率显著增加，提示精子冷冻对胚胎发育有潜在的负面影响。分别从人和小鼠的冻融精子和新鲜精子中检测到21个和95个差异表达的miRNA（DEmiR）。miR-148 b-3 p在人和小鼠冻融精子中均下调，在使用冻存精子受精后的胚胎中也降低。采用实时荧光定量PCR（qRT-PCR）和Western blot（WB）技术，在小鼠胚胎中鉴定miR-148 b-3 p的靶基因Pten。此外，与精子相比，小鼠卵母细胞冷冻保存的共同特征也得到了检测; miR-148 b-3 p在冷冻保存的卵母细胞中表达下调。综上所述，我们的研究表明，冷冻保存精子可以改变miRNAs的表达，特别是miR-148 b-3 p在人类和小鼠中的表达，并可能通过增加Pten的表达进一步影响受精和胚胎发育。此外，冷冻保存诱导的miR-148 b-3 p下调在小鼠配子中是保守的。
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.
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通讯作者：
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