玻璃化冷冻牛GV卵母细胞全基因组甲基化模式初探

doi:10.11843/j.issn.0366-6964.2020.05.014

Abstract

Abstract: The aim of this study was to investigate the effect of vitrification on the genomic methylation patterns of bovine GV oocytes. Fresh and vitrified GV oocytes were collected. Single-cell whole-genome methylation sequencing(ScWGBS) technology was used to detect the whole-genome methylation levels of the fresh and vitrified bovine GV oocytes, in order to reveal the difference of DNA methylation patterns between the fresh and vitrified bovine GV oocytes. The results showed that vitrification didn't significantly affect the whole-genome methylation level of bovine GV oocytes. Gene ontology (GO) and kyoto encyclopedia of genes and genomes (KEGG) were used to analyze 140 differentially methylated regions(DMRs). It was found that DMRs were mainly involved in cell development, cytoskeleton organization, and they were mainly enriched in the PI3K-Akt and GnRH signaling pathways, and the partial related genes were screened about oocyte maturation (TSC2), cytoskeleton (NUDC), cell viability (MAFK) etc.. These results can provide the information foundation and research direction for improving the efficiency of vitrified GV oocytes.

Key words: vitrification, GV oocytes, ScWGBS, bovine, DMR

CLC Number:

S823.3

ZHANG Peipei, WANG Jingjing, HAO Haisheng, DU Weihua, PANG Yunwei, QUAN Guobo, ZHAO Shanjiang, ZOU Huiying, HAO Tong, ZHU Huabin, ZHAO Xueming. Study on Whole Genome Methylation Pattern in Vitrified Bovine GV Oocytes[J]. Acta Veterinaria et Zootechnica Sinica, 2020, 51(5): 1030-1039.

References 48

[1]	LEI T,GUO N,LIU J Q,et al.Vitrification of in vitro matured oocytes:effects on meiotic spindle configuration and mitochondrial function[J].Int J Clin Exp Pathol,2014,7(3):1159-1165.
[2]	FADINI R,DAL CANTO M,MIGNINI RENZINI M,et al.Embryo transfer following in vitro maturation and cryopreservation of oocytes recovered from antral follicles during conservative surgery for ovarian cancer[J].J Assist Reprod Genet,2012,29(8):779-781.
[3]	YAMANAKA K I,AONO N,YOSHIDA H,et al.Cryopreservation and in vitro maturation of germinal vesicle stage oocytes of animals for application in assisted reproductive technology[J].Reprod Med Biol,2007, 6(2):61-68.
[4]	EGERSZEGI I,SOMFAI T,NAKAI M,et al.Comparison of cytoskeletal integrity,fertilization and developmental competence of oocytes vitrified before or after in vitro maturation in a porcine model[J]. Cryobiology, 2013,67(3):287-292.
[5]	AMOUSHAHI M,SALEHNIA M,MOWLA S J.Vitrification of mouse MII oocyte decreases the mitochondrial DNA copy number,TFAM gene expression and mitochondrial enzyme activity[J].J Reprod Infertil, 2017,18(4):343-351.
[6]	RAO B S,MAHESH Y U,CHARAN K V,et al.Effect of vitrification on meiotic maturation and expression of genes in immature goat cumulus oocyte complexes[J].Cryobiology,2012,64(3):176-184.
[7]	MA Y S,PAN B,YANG H X,et al.Expression of CD9 and CD81 in bovine germinal vesicle oocytes after vitrification followed by in vitro maturation[J].Cryobiology,2018,81:206-209.
[8]	XIA W,YE S J,ZENG W B,et al.Cytoskeleton genes expression and survival rate comparison between immature and mature yak oocyte after OPS vitrification[J].Anim Biotechnol,2018,29(4):247-251.
[9]	SLIEKER R C,ROOST M S,VAN IPEREN L,et al.DNA methylation landscapes of human fetal development[J]. PLoS Genet,2015,11(10):e1005583.
[10]	LI E.Chromatin modification and epigenetic reprogramming in mammalian development[J].Nat Rev Genet,2002,3(9):662-673.
[11]	LIANG Y,FU X W,LI J J,et al.DNA methylation pattern in mouse oocytes and their in vitro fertilized early embryos:effect of oocyte vitrification[J].Zygote,2014,22(2):138-145.
[12]	VAJTA G,HOLM P,KUWAYAMA M,et al.Open pulled straw (OPS) vitrification:a new way to reduce cryoinjuries of bovine ova and embryos[J].Mol Reprod Dev,1998,51(1):53-58.
[13]	SMALLWOOD S A,LEE H J,ANGERMUELLER C,et al.Single-cell genome-wide bisulfite sequencing for assessing epigenetic heterogeneity[J].Nat Methods,2014,11(8):817-820.
[14]	BOLGER A M,LOHSE M,USADEL B.Trimmomatic:a flexible trimmer for Illumina sequence data[J]. Bioinformatics,2014,30(15):2114-2120.
[15]	KRUEGER F,ANDREWS S R.Bismark:a flexible aligner and methylation caller for Bisulfite-Seq applications[J]. Bioinformatics,2011,27(11):1571-1572.
[16]	GUO H S,ZHU P,WU X L,et al.Single-cell methylome landscapes of mouse embryonic stem cells and early embryos analyzed using reduced representation bisulfite sequencing[J].Genome Res,2013,23(12):2126-2135.
[17]	KANEHISA M,ARAKI M,GOTO S,et al.KEGG for linking genomes to life and the environment[J].Nucleic Acids Res,2008,36(S1):D480-D484.
[18]	SENDŽIKAITĖ G,KELSEY G.The role and mechanisms of DNA methylation in the oocyte[J].Essays Biochem,2019,63(6):691-705.
[19]	FANG Y,ZHANG X S,ZHANG J L,et al.Global DNA methylation and related mRNA profiles in sheep oocytes and early embryos derived from pre-pubertal and adult donors[J].Anim Reprod Sci,2016, 164:144-151.
[20]	FARLIK M,SHEFFIELD N C,NUZZO A,et al.Single-cell DNA methylome sequencing and bioinformatic inference of epigenomic cell-state dynamics[J].Cell Rep,2015,10(8):1386-1397.
[21]	LI Q N,GUO L,HOU Y,et al.The DNA methylation profile of oocytes in mice with hyperinsulinaemia and hyperandrogenism as detected by single-cell level whole genome bisulphite sequencing (SC-WGBS) technology[J].Reprod Fertil Dev,2018,30(12):1713-1719.
[22]	YU B,DONG X,GRAVINA S,et al.Genome-wide,single-cell DNA methylomics reveals increased non-CpG methylation during human oocyte maturation[J].Stem Cell Rep,2017,9(1):397-407.
[23]	赵亚涵,郝海生,杜卫华,等.新鲜、玻璃化冷冻牛卵母细胞体外受精囊胚全基因组甲基化模式初探[J].畜牧兽医学报,2019,50(6):1179-1188.ZHAO Y H,HAO H S,DU W H,et al.Study on whole genome methylation pattern of in vitro fertilized blastocysts from fresh and vitrified bovine oocytes[J].Acta Veterinaria et Zootechnica Sinica,2019,50(6):1179-1188.(in Chinese)
[24]	GRAVINA S,DONG X,YU B,et al.Single-cell genome-wide bisulfite sequencing uncovers extensive heterogeneity in the mouse liver methylome[J].Genome Biol,2016,17(1):150.
[25]	ADHIKARI D,FLOHR G,GORRE N,et al.Disruption of Tsc2 in oocytes leads to overactivation of the entire pool of primordial follicles[J].Mol Hum Reprod,2009,15(12):765-770.
[26]	PIKIOU O,VASILAKI A,LEONDARITIS G,et al.Effects of metformin on fertilisation of bovine oocytes and early embryo development:possible involvement of AMPK3-mediated TSC2 activation[J]. Zygote, 2015,23(1):58-67.
[27]	ZHANG C,ZHANG W,LU Y,et al.NudC regulates actin dynamics and ciliogenesis by stabilizing cofilin 1[J].Cell Res,2016,26(2):239-253.
[28]	WANG R,ZHENG J,ZHANG D S,et al.Wnt1-induced MAFK expression promotes osteosarcoma cell proliferation[J].Genet Mol Res,2015,14(3):7315-7325.
[29]	GUTNISKY C,MORADO S,GADZE T,et al.Morphological,biochemical and functional studies to evaluate bovine oocyte vitrification[J].Theriogenology,2020,143:18-26.
[30]	GUO X F,YU X L,ZHANG F,et al.Effect of liquid helium vitrification on cytoskeleton of immature cattle oocytes[J].Anim Reprod Sci,2017,187:91-99.
[31]	EPPIG J J.Intercommunication between mammalian oocytes and companion somatic cells[J].BioEssays, 1991,13(11):569-574.
[32]	MAO G K,LI J X,BIAN F H,et al.Gap junction-mediated cAMP movement between oocytes and somatic cells[J].Front Biosci (Elite Ed),2013,5:755-767.
[33]	VAN BLERKOM J.Mitochondria in human oogenesis and preimplantation embryogenesis:engines of metabolism,ionic regulation and developmental competence[J].Reproduction,2004,128(3):269-280.
[34]	MOAWAD A R,XU B Z,TAN S L,et al.L-carnitine supplementation during vitrification of mouse germinal vesicle stage-oocytes and their subsequent in vitro maturation improves meiotic spindle configuration and mitochondrial distribution in metaphase II oocytes[J].Hum Reprod,2014,29(10):2256-2268.
[35]	FU X W,SHI W Q,ZHANG Q J,et al.Positive effects of Taxol pretreatment on morphology,distribution and ultrastructure of mitochondria and lipid droplets in vitrification of in vitro matured porcine oocytes[J]. Anim Reprod Sci,2009,115(1-4):158-168.
[36]	RHO G J,KIM S,YOO J G,et al.Microtubulin configuration and mitochondrial distribution after ultra-rapid cooling of bovine oocytes[J].Mol Reprod Dev,2002,63(4):464-470.
[37]	GUO J,NIU Y J,SHIN K T,et al.Fatty acid synthase knockout impairs early embryonic development via induction of endoplasmic reticulum stress in pigs[J].J Cell Physiol,2018,233(5):4225-4234.
[38]	EPPIG J J.Coordination of nuclear and cytoplasmic oocyte maturation in eutherian mammals[J].Reprod Fertil Dev,1996,8(4):485-489.
[39]	LOWTHER K M,WEITZMAN V N,MAIER D,et al.Maturation,fertilization,and the structure and function of the endoplasmic reticulum in cryopreserved mouse oocytes[J].Biol Reprod,2009,81(1):147-154.
[40]	ZHAO X M,HAO H S,DU W H,et al.Melatonin inhibits apoptosis and improves the developmental potential of vitrified bovine oocytes[J].J Pineal Res,2016,60(2):132-141.
[41]	GUO R J,WANG X R,LI Q H,et al.Follicular fluid meiosis-activating sterol (FF-MAS) promotes meiotic resumption via the MAPK pathway in porcine oocytes[J].Theriogenology, 2019,doi:10.1016/j. theriogenology. 2019.11.012.
[42]	PETRUNEWICH M A,TRIMARCHI J R,HANLAN A K L,et al.Second meiotic spindle integrity requires MEK/MAP kinase activity in mouse eggs[J].J Reprod Dev,2009,55(1):30-38.
[43]	SHA Q Q,DAI X X,DANG Y J,et al.A MAPK cascade couples maternal mRNA translation and degradation to meiotic cell cycle progression in mouse oocytes[J].Development,2017,144(3):452-463.
[44]	MAKKER A,GOEL M M,MAHDI A A.PI3K/PTEN/Akt and TSC/mTOR signaling pathways,ovarian dysfunction,and infertility:an update[J].J Mol Endocrinol,2014,53(3):R103-R118.
[45]	WANG L Q,LIU J C,CHEN C L,et al.Regulation of primordial follicle recruitment by cross-talk between the Notch and phosphatase and tensin homologue (PTEN)/AKT pathways[J].Reprod Fertil Dev,2016, 28(6):700-712.
[46]	ANDRADE G M,DA SILVEIRA J C,PERRINI C,et al.The role of the PI3K-Akt signaling pathway in the developmental competence of bovine oocytes[J].PLoS One,2017,12(9):e0185045.
[47]	POPOVIC-TODOROVIC B,SANTOS-RIBEIRO S,DRAKOPOULOS P,et al.Predicting suboptimal oocyte yield following GnRH agonist trigger by measuring serum LH at the start of ovarian stimulation[J].Hum Reprod,2019,34(10):2027-2035.
[48]	ZANETTI B F,BRAGA D P A F,SETTI A S,et al.Effect of GnRH analogues for pituitary suppression on oocyte morphology in repeated ovarian stimulation cycles[J].JBRA Assist Reprod,2020,24(1):24-29.

Study on Whole Genome Methylation Pattern in Vitrified Bovine GV Oocytes

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