Bovine imprinted gene analysis in assisted reproduction techniques

辅助生殖技术中的牛印记基因分析

• 批准号: 524915-2018
• 负责人: Sirard, MarcAndré
• 金额: $ 2.08万
• 依托单位: Université Laval
• 依托单位国家: 加拿大
• 项目类别: Collaborative Research and Development Grants
• 财政年份: 2018
• 资助国家: 加拿大
• 起止时间: 2018-01-01 至 2019-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=665323
• 关键词: Bovine; imprinted; gene; analysis; assisted

The production of bovine embryos through in vitro maturation and fertilization is becoming an important tool of the genomic revolution in dairy cows. The capacity to select embryos based on the father and the mother genotypes and the possibility to do that at the blastocyst level before a pregnancy is initiated is changing the way animals are produced and selected. The molecular analysis of these embryos indicate some differences according to the age of the parent and according to the culture or hormonal environment. Such influences may be temporary but in some case, it may influence the post-natal phenotype suggesting that more research is required to understand the changes observed. This project is based on the hypothesis that some of the methylation patterns are acquired in the last portion of folliculogenesis and could be influenced by the environment created to generate these oocytes. Such differences may not be erased during the first week of culture or sensitive to the conditions during this time. To test this hypothesis, 32 embryos (day 12) obtained from pre-puber donors (8)will be compared to embryos obtained from adults and produced in vitro with (8) or without serum (8) or in vivo (control)(8). The DNA from these embryos will be extracted and bisulfited to reveal methylation marks. The analysis of 10 imprinted genes will then be compared across treatments to assess if the pattern is modified and if all embryos are affected or only a subset. The second part will assess the methylation level of the 4 best imprinted indicator in 48 blastocysts and biopsies to develop a rapid day 7 test permitting the monitoring of any modification to the culture conditions or ovarian stimulation. This work will generate markers to assess the required hormonal and culture environment to minimize epigenetic perturbation in bovine embryos generated by ART.****

通过体外成熟和受精生产牛胚胎正在成为奶牛基因组革命的重要工具。 根据父亲和母亲基因型选择胚胎的能力以及在开始妊娠前在囊胚水平上选择胚胎的可能性正在改变动物的生产和选择方式。这些胚胎的分子分析表明，根据父母的年龄以及文化或激素环境，存在一些差异。 这种影响可能是暂时的，但在某些情况下，它可能会影响产后表型，这表明需要更多的研究来了解观察到的变化。 该项目基于这样的假设：一些甲基化模式是在卵泡发生的最后部分获得的，并且可能受到生成这些卵母细胞所创造的环境的影响。这种差异可能不会在培养的第一周内被消除，或者对这段时间的条件敏感。为了检验这一假设，将从青春期前供体 (8) 获得的 32 个胚胎（第 12 天）与从成人获得的胚胎进行比较，并在体外（8）或无血清（8）或体内（对照）（8）中产生。 这些胚胎的 DNA 将被提取并进行亚硫酸盐处理，以揭示甲基化标记。然后，对 10 个印记基因的分析将在不同的治疗方法中进行比较，以评估模式是否被修改，以及是否所有胚胎都受到影响，或者只有一个子集受到影响。第二部分将评估 48 个囊胚和活检中 4 个最佳印记指标的甲基化水平，以开发快速第 7 天测试，从而监测培养条件或卵巢刺激的任何修改。这项工作将生成标记来评估所需的激素和培养环境，以最大限度地减少 ART 生成的牛胚胎的表观遗传扰动。****