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ID:32824215
大小:7.05 MB
页数:45页
时间:2019-02-16
《igf-1对水牛植入前胚胎发育及凋亡的影响》由会员上传分享,免费在线阅读,更多相关内容在学术论文-天天文库。
1、广西大学硕士学位论文IGF--1对水牛植入前胚胎发育及凋亡的影响姓名:罗婷申请学位级别:硕士专业:动物遗传育种与繁殖指导教师:石德顺;陆凤花201206IGF.1对水牛植入前胚胎发育及凋亡的影响摘’要本研究主要探讨胰岛素样生长因子.1(IGF一1)对水牛体外受精(IⅦ)及体细胞核移植(SCNT)胚胎发育和凋亡的影响及其作用机制,为进一步完善水牛胚胎体外培养体系及提高胚胎质量提供一定的科学依据。本研究分为以下五个试验。试验1通过实时荧光定量PCR的方法研究IGF.1受体(IGF.1R)在植入前水牛IⅦ及SCNT
2、胚胎中的表达情况。结果显示,IGF.1R在水牛IVF及SCNT各时期胚胎中均有表达,但在胚胎分裂的早期(IⅦ:2-cell:SCNT:2.8cell)表达量较高,与卵母细胞的表达量无显著差异(P>o.05),而后逐渐下降,到囊胚期达到最低。试验2主要探讨培养液中添加IGF.1对水牛胚胎发育的影响。结果发现,添加不同浓度的IGF.1(O、5、lO、50、100ng/mL)对水牛IⅦ胚胎的分裂率没有显著影响(P>O.05),但添加50ng/mLIGF一1组胚胎的囊胚发育率显著高于不添加IGF.1的对照组(35.1
3、2%VS22.97%,P<0.05);用添加50ng/mLIGF一1的培养液培养水牛体细胞核移植胚胎,与不添加IGF.1的对照组相比胚胎分裂率差异不显著(P>O.05),但囊胚率得到显著提高(46.43%VS34.18%,P<0.05)。试验3研究IGF一1对水牛IⅥ’及SCNT胚胎IGF一1RmRNA表达的影响时发现,添加50ng/mLIGF一1培养的水牛IVF及SCNT囊胚IGF一1RmRNA的表达量与不添加IGF一1的对照组相比显著升高(P4、细胞凋亡情况的结果显示,添加50ng/mLIGF.1培养液培养的水牛IVF及SCNT胚胎其囊胚细胞数均显著高于不添加IGF一1的对照组(P5、受精及体细胞核移植各细胞期胚胎中都有表达;(2)水牛胚胎培养液中添加50ng/mL的IGF.1能够促进胚胎的发育,增加囊胚细胞数,并降低囊胚细胞凋亡率;(3)IGF一1通过促进IGF一1R的表达,下调促凋亡基因bax,上调抑凋亡基因bcl.2的表达来促进胚胎的发育和降低胚胎的凋亡率。关键词:IGF一1水牛体外受精体细胞核移植凋亡EFFECTSOFINSULIN—LIKEGROWTHFACTOR-I(IGF.1)ONTHEDEVELOPMENTANDAPOPTOSISOFPREIMPLANTATIONBUFFA6、LOEMBRYOSABSTRACTThisstudywastoexploretheeffectsofinsulin—Hkegrowthfactor—I(IGF—1)onthedevelopmentandapoptosisofIVF/SCNTpreimplantationbuffaloembryos,andinvestigatetheinfluencemechanismsofIGF一1onembryonicdevelopmentandapoptosis,inordertoprovidescientificbas7、isforimprovingthequalityofembryosandtheefficiencyofembryosinvitroculturesystem.Fiveexperimentsweredesignedinthisstudy.Theexperiment1,themRNAexpressionlevelofIGF一1RonpreimplantationbuffaloIVF/SCNTembryoswasinvestigatedbyReal-timequantitativePCR.Theresultsind8、icatedthatIGF一1Rwerepresentinalldevelopmentstageofpreimplantationbuffaloembryos,andthemRNAexpressionlevelofIGF一1Rwashigherinearlyembryos(2-cellinIⅦ,2—8cellinSCNT),whichwasnosignificantdifferencewithooc
4、细胞凋亡情况的结果显示,添加50ng/mLIGF.1培养液培养的水牛IVF及SCNT胚胎其囊胚细胞数均显著高于不添加IGF一1的对照组(P5、受精及体细胞核移植各细胞期胚胎中都有表达;(2)水牛胚胎培养液中添加50ng/mL的IGF.1能够促进胚胎的发育,增加囊胚细胞数,并降低囊胚细胞凋亡率;(3)IGF一1通过促进IGF一1R的表达,下调促凋亡基因bax,上调抑凋亡基因bcl.2的表达来促进胚胎的发育和降低胚胎的凋亡率。关键词:IGF一1水牛体外受精体细胞核移植凋亡EFFECTSOFINSULIN—LIKEGROWTHFACTOR-I(IGF.1)ONTHEDEVELOPMENTANDAPOPTOSISOFPREIMPLANTATIONBUFFA6、LOEMBRYOSABSTRACTThisstudywastoexploretheeffectsofinsulin—Hkegrowthfactor—I(IGF—1)onthedevelopmentandapoptosisofIVF/SCNTpreimplantationbuffaloembryos,andinvestigatetheinfluencemechanismsofIGF一1onembryonicdevelopmentandapoptosis,inordertoprovidescientificbas7、isforimprovingthequalityofembryosandtheefficiencyofembryosinvitroculturesystem.Fiveexperimentsweredesignedinthisstudy.Theexperiment1,themRNAexpressionlevelofIGF一1RonpreimplantationbuffaloIVF/SCNTembryoswasinvestigatedbyReal-timequantitativePCR.Theresultsind8、icatedthatIGF一1Rwerepresentinalldevelopmentstageofpreimplantationbuffaloembryos,andthemRNAexpressionlevelofIGF一1Rwashigherinearlyembryos(2-cellinIⅦ,2—8cellinSCNT),whichwasnosignificantdifferencewithooc
5、受精及体细胞核移植各细胞期胚胎中都有表达;(2)水牛胚胎培养液中添加50ng/mL的IGF.1能够促进胚胎的发育,增加囊胚细胞数,并降低囊胚细胞凋亡率;(3)IGF一1通过促进IGF一1R的表达,下调促凋亡基因bax,上调抑凋亡基因bcl.2的表达来促进胚胎的发育和降低胚胎的凋亡率。关键词:IGF一1水牛体外受精体细胞核移植凋亡EFFECTSOFINSULIN—LIKEGROWTHFACTOR-I(IGF.1)ONTHEDEVELOPMENTANDAPOPTOSISOFPREIMPLANTATIONBUFFA
6、LOEMBRYOSABSTRACTThisstudywastoexploretheeffectsofinsulin—Hkegrowthfactor—I(IGF—1)onthedevelopmentandapoptosisofIVF/SCNTpreimplantationbuffaloembryos,andinvestigatetheinfluencemechanismsofIGF一1onembryonicdevelopmentandapoptosis,inordertoprovidescientificbas
7、isforimprovingthequalityofembryosandtheefficiencyofembryosinvitroculturesystem.Fiveexperimentsweredesignedinthisstudy.Theexperiment1,themRNAexpressionlevelofIGF一1RonpreimplantationbuffaloIVF/SCNTembryoswasinvestigatedbyReal-timequantitativePCR.Theresultsind
8、icatedthatIGF一1Rwerepresentinalldevelopmentstageofpreimplantationbuffaloembryos,andthemRNAexpressionlevelofIGF一1Rwashigherinearlyembryos(2-cellinIⅦ,2—8cellinSCNT),whichwasnosignificantdifferencewithooc
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