miR-449b对绵羊早期胚胎发育的影响

doi:10.16431/j.cnki.1671-7236.2022.07.024

Abstract

Abstract: 【Objective】 The study was to investigate the effect of miR-449b on early embryo development in sheep.【Method】 Purified sheep sperm and mature oocytes were obtained by Percoll density gradient centrifugation and in vitro maturation of cumulus-oocyte complexes (COCs) for 16-18 h,respectively.And fibroblasts were isolated from the ear tissues of one-month-old Suffolk sheep.Firstly,the relative expression of miR-449b in sperms,mature oocytes and fibroblasts were compared by Real-time quantitative PCR.Mature oocytes were injected with 0.9% normal saline (Con group),NC mimic (NC mimic group) and miR-449b mimic (miR-449b mimic group) by microinjection,respectively.Then the embryonic cleavage rate and blastocyst rate were counted on the 2nd and 7th day after parthenogenetic activation. Mature oocytes were fertilized in vitro (IVF), 2-cell embryos in IVF, Con, NC mimic and miR-449b mimic groups were collected, and the changes of histone H3K9me3 were compared by immunofluorescence method. 【Result】 The expression of miR-449b in sperm was significantly higher than that in mature oocytes and fibroblasts (P＜0.05). Compared with Con group, the cleavage rates of embryos in miR-449b mimic and NC mimic groups were significantly decreased (P＜0.05), and the cleavage rate of embryos in NC mimic group was significantly lower than that of miR-449b mimic group (P＜0.05). The blastocyst rate of embryos in miR-449b mimic group was increased, but the difference was not significant (P＞0.05). Histone H3K9me3 was expressed in 2-cell embryos in IVF, Con, NC mimic and miR-449b mimic groups, and was expressed in the nucleus. Compared with Con group, the expression level of histone H3K9me3 in 2-cell embryos in NC mimic group was significantly increased (P＜0.05), and was significantly decreased in miR-449b mimic group (P＜0.05). The expression level of histone H3K9me3 in 2-cell embryos in NC mimic group was significantly higher than that in miR-449b mimic group (P＜0.05), but there was no significant difference between miR-449b mimic and IVF groups (P＞0.05). 【Conclusion】 miR-449b could significantly increase the development rate of early sheep embryos and reduce the expression of H3K9me3 in early embryos.

Key words: miR-449b; early embryo; H3K9me3; sperm

CLC Number:

S813.1

REN Xiumeiao, YAO Xudong, MENG Yaqi, GUO Yanhua, TANG Hong, ZHANG Yiyuan, ZHAO Xingwang, WANG Limin, ZHOU Ping. Effects of miR-449b on Early Embryo Development in Sheep[J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(7): 2661-2668.

References

[1] RANDO O J.Daddy issues:Paternal effects on phenotype[J].Cell,2012,151:702-708.
[2] 贾丹.牛精子RNA对早期胚胎发育影响的初步研究[D].杨凌:西北农林科技大学,2016. JIA D.Effect of sperm-borne RNAs on the early development of bovine embryos[D].Yangling:Northwest A & F University,2016.(in Chinese)
[3] LEWIS B P,BURGE C B,BARTEL D P.Conserved seed pairing,often flanked by adenosines,indicates that thousands of human genes are microRNA targets[J].Cell,2005,120(1):15-20.
[4] CARRELL D T.Contributions of spermatozoa to embryogenesis:Assays to evaluate their genetic and epigenetic fitness[J]. Reproductive Biomedicine Online,2008,16(4):474-484.
[5] WANG M,GAO Y,QU P,et al.Sperm-borne miR-449b influences cleavage,epigenetic reprogramming and apoptosis of SCNT embryos in bovine[J].Scientific Reports,2017,7(1):13403.
[6] BAO J,LI D,WANG L,et al.microRNA-449 and microRNA-34b/c function redundantly in murine testes by targeting E2F transcription factor-retinoblastoma protein (E2F-pRb) pathway[J].Journal of Biological Chemistry,2012,287(26):21686-21698.
[7] WU J,ZHU H,SONG W,et al.Identification of conservative microRNAs in Saanen dairy goat testis through deep sequencing[J].Reproduction in Domestic Animals,2014,49:32-40.
[8] WANG C,LIU X,GAO Y,et al.Reprogramming of H3K9me3-dependent heterochromatin during mammalian embryo development[J]. Nature Cell Biology,2018,20(5):12.
[9] OKAMOTO I,OTTE A P,ALLIS C D,et al.Epigenetic dynamics of imprinted X inactivation during early mouse development[J].Science,2004,303:644-649.
[10] FELDMAN N,GERSON A,FANG J,et al.G9a-mediated irreversible epigenetic inactivation of Oct-3/4 during early embryogenesis[J].Nature Cell Biology,2006,8(2):188-194.
[11] FUKUDA A,TOMIKAWA J,MIURA T,et al.The role of maternal-specific H3K9me3 modification in establishing imprinted X-chromosome inactivation and embryogenesis in mice[J].Nature Communications,2014,5(1):5564.
[12] TACHIBANA M.G9a histone methyltransferase plays a dominant role in euchromatic histone H3 lysine 9 methylation and is essential for early embryogenesis[J].Genes & Development,2002,16(14):1779-1791.
[13] MESSERSCHMIDT D M,SOLTER D,KNOWLES B B,et al.Trim28 is required for epigenetic stability during mouse oocyte to embryo transition[J].Science,2012,335:1499-1502.
[14] YUAN S,SCHUSTER A,TANG C,et al.Sperm-borne miRNAs and endo-siRNAs are important for fertilization and preimplantation embryonic development[J].Development,2016,143(4):635-647.
[15] CONINE C,SUN F,SONG L,et al.Small RNAs gained during epididymal transit of sperm are essential for embryonic development in mice[J].Developmental Cell,2018,46(4):470-480.
[16] GRIVNA S T,PYHTILA B,LIN H.MIWI associates with translational machinery and PIWI-interacting RNAs (piRNAs) in regulating spermatogenesis[J].Proceedings of the National Academy of Sciences of the United States of America,2006,103(36):13415-13420.
[17] KRAWETZ S A.Paternal contribution:New insights and future challenges[J].Nature Reviews Genetics,2005,6(8):633-642.
[18] LIN,S,GREGORY R I.microRNA biogenesis pathways in cancer[J].Nature Reviews Cancer,2015,15(6):321-333.
[19] CHENG C J,BAHAL R,BABAR I A,et al.microRNA silencing for cancer therapy targeted to the tumour microenvironment[J].Nature,2015,518(7537):107-110.
[20] 潘虹,陈蓓丽,李腾龑,等.miR-449b多态性位点rs10061133差异调控LHCGR表达[J].生殖医学杂志,2019,28(12):5. PAN H,CHEN B L,LI T Y,et al.miR-449b polymorphism rs10061133 differentially regulates LHCGR expression[J].Journal of Reproductive Medicine,2019,28(12):5.(in Chinese)
[21] LIU W M,PANG R T,CHIU P C,et al.Sperm-borne microRNA-34c is required for the first cleavage division in mouse[J].Proceedings of the National Academy of Sciences of the United States of America,2012,109(2):490-494.
[22] SINHA P B,TESFAVE D,RINGS F,et al.microRNA-130b is involved in bovine granulosa and cumulus cells function,oocyte maturation and blastocyst formation[J].Journal of Ovarian Research,2017,10(1):37.
[23] WANG M,DU Y,GAO S,et al.Sperm-borne miR-202 targets SEPT7 and regulates first cleavage of bovine embryos via cytoskeletal remodeling[J].Development,2021,148(5):1-13.
[24] WANG L,ZHANG J,DUAN J,et al.Programming and inheritance of parental DNA methylomes in mammals[J].Cell,2014,157(4):979-991.
[25] YAMANAKA K I,SUGIMURA S,WAKAI T,et al.Acetylation level of histone H3 in early embryonic stages affects subsequent development of miniature pig somatic cell nuclear transfer embryos[J].Journal of Reproduction and Development,2009,55(6):638-644.
[26] ZHOU L Q,DEAN J.Reprogramming the genome to totipotency in mouse embryos[J].Trends in Cell Biology,2015,25(2):82-91.
[27] YONG J C,LIN C P,RISSO D,et al.Deficiency of microRNAmiR-34a expands cell fate potential in pluripotent stem cells[J].Science,2017,355(6325):581-582.
[28] MACFARLAN T S,GIFFORD W D,DRISCOLL S,et al.Embryonic stem cell potency fluctuates with endogenous retrovirus activity[J].Nature,2012,487(7405):57-63.

Effects of miR-449b on Early Embryo Development in Sheep

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