DISTRIBUTION OF MITOCHONDRIA IN BOVINE OOCYTES & DVMTL COMPETENCE IN VITRO

目前的研究是结合以前的工作进行的 Jayne Squirrell在D4R中完成的， 线粒体，在早期仓鼠胚胎中。 这个假设 牛卵母细胞内线粒体分布 在次优条件下的体外成熟改变， 降低发展能力。 成熟的过程是 非常关键，最终会影响到 使胚胎发育成健康的胚泡和胎儿， 产生后代。 我们想先调查一下 线粒体的分布，在未成熟的卵母细胞收集从 屠宰牛的卵巢。 然后我们将卵母细胞置于 和我们实验室开发的差成熟培养基，然后染色， 成熟的卵母细胞寻找线粒体分布， 微管蛋白和肌动蛋白。 我们假设， 成熟将反映卵母细胞的发育能力 经过体外受精和培养。 这将有助于我们 进一步开发牛卵母细胞的成熟培养基， 具有最大体外发育能力的胚胎。 在此之后 第一个主要项目，我们想调查这些相同的 经阴道超声引导的卵母细胞参数 卵母细胞提取。 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 线粒体的最佳分布 我们还要 将24小时的成熟期分成6小时的间隔， 仔细研究线粒体在成熟过程中的运动 过程 然后，我们将继续研究 差的和好的成熟培养基对所产生的胚胎， 原核期或单细胞期、2细胞期和8细胞期 阶段 我们假设线粒体的改变 在成熟过程中可能发生的分布在结果中持续存在 胚胎，这可能是发展的机制， 暴露了 最后，我们想研究一下这些变化 在单个活卵母细胞和胚胎中，使用双光子显微镜。 我们 我相信这些实验将帮助我们更多地了解 卵母细胞在成熟的关键过程中，以及如何 在成熟过程中发生的扰动影响了 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.