BOVINE EMBRYO SEXING ASSAYS BASED ON MPEFM

即使成功地创造了转基因的、核的 移植，或克隆胚胎，我们仍然面临着同样的几率 发育成雌性和雄性后代。最近的调查显示 建议多光子激发荧光显微镜 (MPEFM)是一种观察发育过程的微创技术。 胚胎。此外，性别特异性标记物，如葡萄糖-6-磷酸 脱氢酶(G6PDH)水平和H-Y抗原表达可以预测 胚胎性行为。基因性别可以用聚合酶链式反应来确认，但这项技术 是侵入性的而且代价高昂。这个项目的总体目标是 开发一种允许快速鉴定研究性别的分析方法 胚胎，最终应用于商业乳制品和 牛肉产业。我们评估这项试验成功与否的方法 将通过调查评估1)发展 用一种新的UPEFM对牛胚胎进行成像，2)检测 NTEFM使用反义技术的X连锁G6PDH消息，3) 卵泡发育过程中H-Y抗原的检测 MPEFM；4)G6PDH胞浆水平与H-Y结合的成功 抗原检测可以可靠地预测遗传性别。这些信息 在这些实验中获得的结果将增加我们的基本知识 使用实时显微镜了解胚胎发育。这个 一种准确预测基因性别的分析方法的发展 在移植前对胚胎进行处理会增加 这些进步都是实现的。此外，这项研究将具有 直接应用于提高玉米繁殖率的研究 家养物种。

Even with the successful creation of a transgenic, nuclear transfer, or cloned embryo, we are still faced with equal odds of developing female and male offspring. Recent investigations have suggested that multiple-photon excitation fluorescence microscopy (MPEFM) is a minimally invasive technique for observing the developing embryo. Furthermore, sex-specific markers such glucose-6-phosphate dehydrogenase (G6PDH) levels and H-Y antigen expression can predict embryo sex. Genetic sex can be confirmed using PCR, but the technique is invasive and costly. The overall goal of this project is to develop an assay which would allow for the rapid sexing of research embryos, with the eventual application to the commercial dairy and beef industries. Our approach for assessing the success of this assay will come through investigations evaluating 1) the development of bovine embryos imaged with a novel UPEFM, 2) the detection of the X-linked G6PDH message using antisense technology by NTEFM, 3) the detection of H-Y antigen on developing morulae and blastocysts using MPEFM, and 4) the success of combining G6PDH cytosolic levels and H-Y antigen detection to reliably predict genetic sex. The information obtained during these experiments will add to our fundamental knowledge of embryo development using real-time microscopy. The development of an assay to correctly predict the sex of a genetically manipulated embryo prior to transfer will increase the rate at which these advances are realized. Furthermore, this research will have direct application in improving the reproductive efficiency in domestic species.