兩步體外成熟法對(duì)綿羊卵母細(xì)胞發(fā)育能力的影響

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1、河北農(nóng)業(yè)大學(xué)碩士學(xué)位論文兩步體外成熟法對(duì)綿羊卵母細(xì)胞發(fā)育能力的影響姓名：胡媛媛申請(qǐng)學(xué)位級(jí)別：碩士專業(yè)：動(dòng)物遺傳育種與繁殖指導(dǎo)教師：田樹軍20120527摘要體外成熟綿羊卵母細(xì)胞隨后的胚胎發(fā)育能力遠(yuǎn)低于體內(nèi)成熟的卵母細(xì)胞，其主要原因之一是卵母細(xì)胞的細(xì)胞核成熟早于細(xì)胞質(zhì)成熟，即細(xì)胞核與細(xì)胞質(zhì)成熟不同步。本文以綿羊卵母細(xì)胞為材料，利用幾種核成熟抑制劑組合對(duì)其進(jìn)行兩步體外成熟，研究延遲其細(xì)胞核成熟進(jìn)程，目的是提高體外成熟卵母細(xì)胞及后期胚胎的發(fā)育能力。首先，采用不同濃度西洛酰胺(Cilo蛐idc)延遲卵母細(xì)胞核成熟進(jìn)程、促

2、卵泡素(FSH)對(duì)Cilostamide抑制卵母細(xì)胞核成熟，以及Cilostamide抑制卵母細(xì)胞核成熟對(duì)隨后胚胎發(fā)育能力的影響進(jìn)行研究。結(jié)果表明：Cilostamide對(duì)綿羊卵母細(xì)胞核成熟進(jìn)程的抑制具有劑量依賴性，5pmol／L的Cilostamide為抑制卵母細(xì)胞核成熟的適宜濃度，可使綿羊卵母細(xì)胞的核成熟進(jìn)程延長約4h，而高濃度(10pmol／L和20“mol／L)的Cilostamide將導(dǎo)致部分卵母細(xì)胞發(fā)生不可逆性抑制；FSH與Cilostamide對(duì)綿羊卵母細(xì)胞的核成熟起協(xié)同抑制作用，成熟液中含有Cil

3、ostamide時(shí)的卵母細(xì)胞體外成熟率隨著FSH濃度增加呈現(xiàn)下降趨勢(shì)；綿羊卵母細(xì)胞在含有5lunol／L的Cilostamide的成熟液中體外成熟28h，然后進(jìn)行化學(xué)激活的囊胚發(fā)育率達(dá)至U(40．4％)，顯著高于對(duì)照組(29．0％)儼

4、Itmol／LIBMX+100vmol／LFSK或100I_unol／LIBMX+50Imml／LFSK，囊胚發(fā)育率分別達(dá)至lJ(51．6％和49．8％)，均顯著高于對(duì)照組(33．1％)伊

5、P水平已降至較低而實(shí)驗(yàn)組卵母細(xì)胞中cAMP仍然保持較高水平。到30h時(shí)實(shí)驗(yàn)組與對(duì)照組卵母細(xì)胞內(nèi)cAMP水平相比，差異不顯著paos)。關(guān)鍵詞：3一異丁基．1．甲基黃嘌呤；毛喉素；西洛酰胺；卵母細(xì)胞；體外成熟；綿羊TheEffectofTwo-stepMaturationinvitroforDevelopmentalCapacityofOocytesfromSheepAuthor：Huyu躚yuanAdvisor：TianShujunMajor．AnimalGenetics，BreedingandReproduct

6、ionAbstractAtpresent,thesubsequentembryodevelopmentabilityofovineoocytesmaturatedinvitroisfarlowerthantheembryosderivedfrominvivomaturedoocytes,themainreasonmaybethenuclearmaturationearlierthancytoplasmicmaturation．Itistosayoocytenuclearandcytoplasmicmaturati

7、onisnotsynchronous．Inthispaper，ovineoocyteswerematuredbytwostepmaturemethodwhichusingavarietyofnuclearmaturationinhibitors，delayedoocytenuclearmaturation，inordertomaketheoocytenuclearandcytoplasmicmatured必synchronousaspossible，SOastoimprovethequalityofoocytes

8、maturedinvitroandsubsequentembryodevelopmentcompetence．Atfirst,researchedthedifferentconcentrationsofCilostamidedelayedoocytenuclearmaturationtimeandinhibiteffect，theinfluenceofFolliclest