DISTRIBUTION OF MITOCHONDRIA IN BOVINE OOCYTES & DVMTL COMPETENCE IN VITRO

目前的研究是与以前的工作一起进行的 在D4R中由Jayne Squirrell完成，涉及分发 线粒体，在仓鼠早期胚胎中。这是一个假设 研究表明，牛卵母细胞内线粒体的分布 在次优条件下的体外成熟与 发展能力下降。成熟的过程是 非常关键，并最终会影响到 结果胚胎发育成一个健康的胚泡和胎儿，以及 繁衍后代。我们想首先调查一下 未成熟卵母细胞中线粒体的分布 被屠宰的牛的卵巢。然后我们会让卵母细胞 和我们实验室开发的低成熟培养液，然后染色 成熟的卵母细胞来寻找线粒体的分布和 微管蛋白和肌动蛋白。我们假设之后的分布 成熟程度将反映卵母细胞的发育能力。 经过体外受精和培养后。这将有助于我们 牛卵母细胞成熟培养液的进一步开发 具有最大体外发育能力的胚胎。在这之后 第一个重大项目，我们想调查这些相同的 经阴道超声引导获取卵母细胞的参数 卵母细胞取回。这些卵母细胞直接从 活牛的卵巢，并将为我们提供进一步的信息 至于线粒体的最佳分布。我们也想要 将24小时的成熟期分为6小时间隔，并 仔细研究线粒体在成熟过程中的运动 进程。然后，我们愿意继续调查 较差和较好的成熟培养液对所产生的胚胎，在 原核，或一个细胞阶段，在2细胞阶段，在8细胞阶段 舞台。我们假设线粒体的变化 在成熟过程中可能发生的分布持续存在于由此产生的 胚胎，这可能是发育是 妥协了。最后，我们想研究一下随着时间的推移这些变化 在单个活的卵母细胞和胚胎中，使用双光子显微镜。我们 相信这些实验将帮助我们更多地了解发生了什么 在卵母细胞成熟的关键过程中，以及如何 在成熟过程中发生的干扰会影响到 由此产生的胚胎。

The current study was developed in conjunction with previous work done in the D4R by Jayne Squirrell involving the distribution of mitochondria, in early stage hamster embryos. The hypothesis of this study is that mitochondrial distribution in bovine oocytes after in vitro maturation in suboptimal conditions is altered consistent with reduced developmental competence. The process of maturation is extremely critical, and can ultimately affect the ability of the resulting embryo to develop into a healthy blastocyst and fetus, and give rise to offspring. We would like to first investigate the distribution of mitochondria, in immature oocytes collected from the ovaries of slaughtered cattle. We will then subject oocytes to good and poor maturation medium developed in our laboratory, then stain the matured oocytes to look for mitochondrial distribution and also for tubulin and actin. We hypothesize that the distribution after maturation will reflect the developmental capability of the oocyte after in vitro fertilization and culture. This will assist us in further developing maturation medium for bovine oocytes to produce embryos with maximal developmental competence in vitro. After this first major project, we would like to investigate these same parameters with oocytes derived from transvaginal ultrasound guided oocyte retrieval. These oocytes are aspirated directly from the ovaries of living cattle, and will provide us with further information as to the optimal mitochondrial distribution. We would also like to divide the maturation period of 24 hours into 6 hour intervals and closely study the movements of mitochondria during the maturation process. We would then like to continue to investigate the effects of poor and good maturation medium on the resulting embryos, at the pronuclear, or one cell stage, at the 2 cell stage, and at the 8 cell stage. We hypothesize that the alterations in mitochondrial distribution that may occur during maturation persist in the resulting embryo and that this may be the mechanism whereby development is compromised. Lastly, we would like to examine these changes over time in a single living oocyte and embryo, using 2 photon microscopy. We believe these experiments will help us learn more about what happens to an oocyte during the critical process of maturation, and how perturbations which occur during maturation effect the development of the resulting embryo.