GDF-9基因真核表达载体的构建及其对牛卵丘细胞扩展相关基因表达的影响

doi:10.16431/j.cnki.1671-7236.2015.06.001

Abstract

Abstract: The objective of the study was to construct the eukaryotic expression vector of bovine GDF-9 gene,and investigate its effect on the expression of expansion-related genes in transfected bovine cumulus cells.Eukaryotic expression vector plasmid pcDNA4 myc-his-GDF-9 was constructed by insert GDF-9 in the multiple cloning site.pcDNA4 myc-his-GDF-9 was transfected into bovine cumulus cells by liposome.The expression of GDF-9 gene was detected by Western blotting and the expression of the expansion-related genes in bovine cumulus cells was detected by qRT-PCR.The results showed that GDF-9 could express in transfected bovine cumulus cells and the expression of expansion-related genes PTGS2,PTX3 and HAS2 in transfected bovine cumulus cells were significantly increased (P< 0.05).In a conclusion,the bovine pcDNA4 myc-his-GDF-9 could effectively transfected into bovine cumulus cells,which could help to study farther functions of the GDF-9 gene.

Key words: bovine; GDF-9; eukaryotic expression vector; expansion-related genes

CLC Number:

LI Rui-lan, OUYANG Xiao-qing, ZHANG Chun-qiang, ZHANG Tong, FAN Xiao-mei, ZHANG Jia-xin. Construction of Eukaryotic Expression Vector of GDF-9 Gene and its Effect on the Expression of Expansion-related Genes in Bovine Cumulus Cells[J]. , 2015, 42(6): 1325-1332.

References

[1] Mcpheron A C,Lee S J.GDF-3 and GDF-9:Two new members of the transforming growth factor-β super family containing a novel pattern of cysteines[J].J Biol Chem,1993,268(5):3444-3449.

[2] Incerti B,Dong J,Borsani G,et al.Structure of the mouse growth/differentiation factor 9 gene[J].J Biochem Biophy Acta,1994,1222(1):125-128.

[3] Elvin J A,Yan C,Matzuk M M.Oocyte-expressed TGF-beta superfamily members in female fertility[J].Mol Cell Endocrinol,2000,159(1-2):1-5.

[4] Knight P G,Glister C.Local roles of TGF-beta superfamily members in the control of ovarian follicle development[J].Anim Reprod Sci,2003,78(3-4):165-183.

[5] Pennetier S,Uzbekova S,Perreau C,et al.Spatio-temporal expression of the germ cell marker genes MATER,ZAR1,GDF9,BMP15 and VASA in adult bovine tissues,oocytes and preim-plantation embryos[J].Biology of Reproduction,2004,71(4):1359-1366.

[6] 梁春年,闫萍,郭贤,等.牛繁殖性状候选基因的研究进展[J].中国畜牧兽医,2011,38(8):103-106.

[7] Kaivo-Oja N,Bondestam J,Kamarainen M,et al.Growth differentiation factor-9 induces Smad-2 activation and inhibin-B production in cultured human granulose-luteal cells[J].J Clin Endocrinol Metab,2003,88(2):755-762.

[8] Wang J,Roy S K.Growth differentiation factor-9 and stem cell factor promote primordial follicle formation in the hamster:Modulation by follicle-stimulating hormone[J].Biol Reprod,2004,70(3):577-585.

[9] Eppig J J,Wigglesworth K,Pendola F,et al.Murine oocytes suppress expression of luteinizing hormone receptor messenger ribonucleic acid by granulosa cells[J].Biol Reprod,1997,56:976-984.

[10] Hussein T S,Froiland D A,Amato F,et al.Oocytes prevent cumulus cell apoptosis by maintaining a morphogenic paracrine gradient of bone morphogenetic proteins[J].J Cell Sci,2005,118:5257-5268.

[11] Paradis F,Moore H S,Pasternak J A,et al.Pig preovulatory oocytes modulate cumulus cell protein and gene expression in vitro[J].Mol Cell Endocrinol,2010,320:87-96.

[12] Regassa A,Rings F,Hoelker M,et al.Transcriptome dynamics and molecular cross-talk between bovine oocyte and its companion cumulus cells[J].BMC Genomics,2011,12:718-757.

[13] Gilchrist R B,Ritter L J,Myllymaa S,et al.Molecular basis of oocyte-paracrine signalling that promotes granulosa cell proliferation[J].J Cell Sci,2006,119:3811-3821.

[14] Li J J,Sugimura S,Mueller T D,et al.Modifications of human growth differentiation factor 9 to improve the generation of embryos from low competence oocytes[J].Mol Endocrinol,2014,29(1):40-52.

[15] Yeo C X,Gilchrist R B,Thompson J G,et al.Exogenous growth differentiation factor 9 in oocyte maturation media enhances subsequent embryo development and fetal viability in mice[J].Hum Reprod,2008,23(1):67-73.

[16] Romaguera R,Morató R,Jiménez-Macedo A R,et al.Oocyte secreted factors improve embryo developmental competence of COCs from small follicles in prepubertal goats[J].Theriogenology,2010,74(6):1050-1059.

[17] 周泽晓,冉雪琴,王嘉福.贵州白山羊GDF9基因编码区1007位点的多态性[J].中国畜牧兽医,2009,36(12):73-76.

[18] Simpson C M,Robertson D M,Al-Musawi S L,et al.Aberrant GDF9 expression and activation are associated with common human ovarian disorders[J].J Clin Endocrinol Metab,2014,99(4):615-624.

[19] 汤展毅,严云勤,高学军,等.牛Myf6基因真核表达载体的构建及在成肌细胞中的表达[J].中国农业科学,2010,43(13):2793-2799.

[20] 白志军,张丽芸,林佐贤,等.pcDNA4/His C-MBL表达载体的构建及其在CHO细胞中的表达[J].第一军医大学学报,2004,24(8):859-863.

[21] 魏晓丽,施桥发,李虹,等.鼠-防御素2(mBD2)真核表达质粒的构建及稳定表达株的鉴定[J].细胞与分子免疫学杂志,2007,23(1):28-31.

[22] Fülöp C,Szántó S,Mukhopadhyay D,et al.Impaired cumulus mucification and female sterility in tumor necrosis factor-induced protein-6 deficient mice[J].Development,2003,130(10):2253-2261.

[23] Russell D L,Robker R L.Molecular mechanisms of ovulation:Co-ordination through the cumulus complex[J].Hum Reprod Update,2007,13:289-312.

[24] Chen L,Russell P T,Larsen W J.Functional significance of cumulus expansion in the mouse:Roles for the preovulatory synthesis of hyaluronic acid with in the cumulus mass[J].Mol Reorod Dev,1993,34(1):87-93.

[25] Fulop C,Salustri A,Hascall V C.Coding sequence of a hyaluronan synthase homologue expressed during expansion of the mouse cumulus-oocyte complex[J].Arch Biochem Biophys,1997,337:261-266.

[26] Salustri A,Garlanda C,Hirsch E,et al.PTX3 plays a key role in the organization of the cumulus oophorus extracellular matrix and in in vivo fertilization[J].Development,2004,131(7):1577-1586.

[27] Varani S,Elvin J A,Yan C,et al.Knockout of pentraxin-3,a downstream target of growth differentiation factor-9,causes female subfertility[J].Mol Endocrinol,2002,16(6):1154-1167.

[28] Elvin J A,Yan C,Matzuk M M.Growth differentiation factor-9 stimulates progesterone synthesis ingranulosa cells via a prostaglandin E2/EP2 receptor pathway[J].Proc Natl Acad Sci USA,2000,97(18):10288-10293.

[29] Lim H,Paria B C,Das S K,et al.Multiple female reproductive failures in cyclooxygenase 2-deficient mice[J].Cell,1997,91(2):197-208.

[30] Gilchrist R B,Lane M,Thompson J G.Oocyte-secreted factors:Regulators of cumulus cell function and oocyte quality[J].Hum Reprod Update,2008,14(2):159-177.

[31] Lee B C,Gomez M N L,Kang J T,et al.Effect of oocyte-secreted factors on porcine in vitro maturation,cumulus expansion and developmental competence of parthenotes[J].Zygote,2012,20(2):135-145.

[32] 欧阳效晴,张春强,栗瑞兰,等.生长分化因子-9对牛卵丘细胞增殖的影响[J].中国细胞生物学报,2014,36(7):920-926.

[33] Lin Z L,Li Y H,Xu Y N,et al.Effects of growth differentiation factor 9 and bone morphogenetic protein 15 on the in vitro maturation of porcine oocytes[J].Reproduction in Domestic Animals,2014,49:219-227.

[34] Varnosfaderani S R,Ostadhosseini S,Hajian M,et al.Importance of the GDF9 signaling pathway on cumulus cell expansion and oocyte competency in sheep[J].Theriogenology,2013,80:470-478.

[35] Sugimura S,Ritter L J,Sutton-McDowall M L,et al.Amphiregulin co-operates with bone morphogenetic protein 15 to increase bovine oocyte developmental competence:Effects on gap junction-mediated metabolite supply[J].Mol Hum Reprod,2014,20(6):499-513.

Construction of Eukaryotic Expression Vector of GDF-9 Gene and its Effect on the Expression of Expansion-related Genes in Bovine Cumulus Cells

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