本文選題：綿羊卵母細(xì)胞 + 超數(shù)排卵��； 參考：《河南農(nóng)業(yè)大學(xué)》2015年碩士論文

【摘要】：本試驗(yàn)研究了綿羊幼羔超數(shù)排卵和成年羊卵母細(xì)胞的體外成熟、孤雌激活以及體細(xì)胞核移植胚胎發(fā)育的特點(diǎn),以期優(yōu)化綿羊超排、卵成熟、卵活化、胚胎發(fā)育條件,為開展發(fā)育機(jī)理研究及提高綿羊克隆效率提供進(jìn)一步的技術(shù)支持。本試驗(yàn)研究結(jié)果主要包括以下幾個(gè)方面:(1)本試驗(yàn)采用寧波、中科院和加拿大激素分別對幼羔進(jìn)行超排處理,對比不同激素對幼羔超排效果的影響。其可用COCs百分比分別為69.6%、87.9%、88.4%,平均可用卵數(shù)分別為15.1枚、25.1枚、27.1枚,差異顯著(P0.05),說明寧波產(chǎn)激素超排效果不如中科院和加拿大激素,而中科院和加拿大之間的可用COCs百分比和平均可用卵數(shù)差異不顯著(P0.05),說明中科院和加拿大激素使用效果相差不大,但是加拿大激素可用COCs百分比和平均可用卵數(shù)比中科院激素分別高0.5%和2枚,說明加拿大激素效果最好。(2)添加FBS培養(yǎng)的卵母細(xì)胞,幼羔組和成年羊組的成熟率分別為51.1%和77.3%,兩者差異顯著(P0.05),添加ESS培養(yǎng)的卵母細(xì)胞,幼羔組和成年羊組分別為59.0%和78.9%,兩者差異顯著(P0.05),說明成年羊卵母細(xì)胞的體外成熟效果要優(yōu)于幼羔。添加ESS的幼羔組成熟率比添加FBS組高7.9%,兩者差異顯著(P0.05),說明添加ESS能夠提高超排羔羊卵母細(xì)胞的體外成熟率。(3)比較Ionomycin+6-DMAP和A23187+(CHX和CB)兩種激活方式,幼羔一和幼羔二組獲得的卵裂率和囊胚率分別為50.7%、13.4%和48.0%、11.1%,兩組的卵裂率和囊胚率差異皆不顯著(P0.05),成年羊一和成年羊二組的卵裂率和囊胚率分別為69.5%、29.5%和67.6%、28.6%,兩組間卵裂率和囊胚率差異不顯著(P0.05),但是,成年羊一組的卵裂率和囊胚率比成年羊二組分別高出1.9%和0.9%,幼羔一組的卵裂率和囊胚率比幼羔二組分別高出2.7%和2.3%,而成年羊組卵裂率和囊胚率皆顯著高于幼羔組(P0.05),說明兩種激活方式都能有效激活綿羊卵母細(xì)胞,而成年羊卵母細(xì)胞激活后發(fā)育效果要比幼羔卵母細(xì)胞好。(4)比較體外培養(yǎng)22~24h后幼羔卵母細(xì)胞和成年羊卵母細(xì)胞去核效果,其去核個(gè)數(shù)、去核成功率分別為72、75、60.0%、50.3%,幼羔卵母細(xì)胞去核成功率比成年羊卵母細(xì)胞高9.7%,兩者差異顯著(P0.05),說明體外成熟22~24h的幼羔卵母細(xì)胞去核效果優(yōu)于成年羊卵母細(xì)胞。(5)幼羔組和成年羊組重構(gòu)胚的卵裂率和囊胚率分別為29.0%、5.0%、58.8%、17.0%,成年羊組卵裂率和囊胚率比幼羔組分別高出29.8%和12%,差異顯著(P0.05),說明幼羔超排卵母細(xì)胞核移植重構(gòu)胚發(fā)育效果不如成年羊卵母細(xì)胞。
[Abstract]:The characteristics of ovulation, parthenogenetic activation and somatic nuclear transfer embryo development of ovulation and adult sheep oocytes were studied in order to optimize the conditions of ovulation, ovum maturation, oocyte activation and embryo development. It provides further technical support for the study of development mechanism and the improvement of sheep cloning efficiency. The results of this study mainly include the following aspects: 1) the effects of different hormones on the superovulation of young lambs were studied by using Ningbo, Chinese Academy of Sciences and Canadian hormones respectively. The percentages of available COCs were 69.6% and 87.9%, respectively, and the average number of available eggs was 15.1 eggs. The difference was significant (P 0.05), which indicated that the effect of hormone superovulation in Ningbo was not as good as that of Chinese Academy of Sciences and Canadian hormone. However, there was no significant difference in the percentage of available COCs and the average number of available eggs between the Chinese Academy of Sciences and Canada, indicating that there was no significant difference in the effect of hormone use between the Chinese Academy of Sciences and Canada. However, the percentage of available COCs and the average number of available eggs in Canada were 0.5% and 2 higher than those in the Chinese Academy of Sciences, respectively, indicating that Canadian hormone had the best effect on oocytes cultured with FBS. The maturation rates of young lambs and adult sheep were 51.1% and 77.3%, respectively. The difference between the two groups was significant (P 0.05). 59.0% and 78.9% of young lambs and adult lambs had significant difference (P0.05%, respectively), which indicated that the maturation effect of adult sheep oocytes was better than that of young lambs. The maturation rate of lambs supplemented with ESS was 7.9 higher than that with FBS. The difference between the two groups was significant (P 0.05), which indicated that the addition of ESS could increase the maturation rate of oocytes of superovulation lambs in vitro.) compared with Ionomycin 6-DMAP, A23187 CHX and CB3, the in vitro maturation rate of oocytes in superovulation lambs could be increased by adding ESS. The cleavage rate and blastocyst rate were 50.7% and 48.0%, respectively. There was no significant difference in cleavage rate and blastocyst rate between the two groups. The cleavage rate and blastocyst rate of adult sheep group 1 and adult sheep group 2 were 69.529.5% and 67.67.628%, respectively. There was no significant difference between blastocyst rate and blastocyst rate, however, The cleavage rate and blastocyst rate of the adult sheep group were 1.9% and 0.9% higher than those of the adult sheep group, respectively. The cleavage rate and blastocyst rate of the lambs group 1 were 2.7% and 2.3% higher than those of the young lambs group, respectively, while the cleavage rate and blastocyst rate of the adult sheep group were significantly higher than those of the young lambs group. P0. 05, which shows that both activation modes can activate ovine oocytes effectively. The developmental effect of adult sheep oocytes after activation was better than that of young lamb oocytes. The success rate of oocyte denucleation in young lambs was higher than that in adult sheep oocytes by 9.7%, and the difference was significant (P0.05), indicating that the oocyte denucleation effect in vitro was better than that in adult sheep oocyte group and adult goat oocyte group (P < 0.05), indicating that the effect of oocyte denucleation in vitro was better than that in adult sheep oocyte group and adult goat oocyte group (P < 0.05). The cleavage rate and blastocyst rate of reconstructed embryos were 29.0% and 5.0%, respectively. The cleavage rate and blastocyst rate of adult sheep group were 29.8% and 12.1% higher than that of young lambs group, respectively. The difference was significant (P 0.05), which indicated that the effect of nuclear transfer of superovulation mother cell of young lambs on embryo reconstruction was not as good as that of adult goat oocytes.
【學(xué)位授予單位】：河南農(nóng)業(yè)大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2015
【分類號】：S826

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