BOVINE EMBRYO SEXING ASSAYS BASED ON MPEFM

即使成功地创造了一个转基因，核 移植，或克隆胚胎，我们仍然面临着同样的机会， 正在发育雌性和雄性后代。 最近的调查 多光子激发荧光显微镜 （MPEFM）是一种微创技术，用于观察发育中的 胚胎 此外，性别特异性标志物，如葡萄糖-6-磷酸， 脱氢酶（G6 PDH）水平和H-Y抗原表达可以预测 胚胎性别。 基因性别可以用PCR来确认，但这项技术 是侵入性的并且成本高昂。 该项目的总体目标是 开发一种检测方法，可以快速确定研究对象的性别 胚胎，最终应用于商业乳品， 牛肉产业。 我们评估该试验成功的方法 将通过调查评估1）发展 牛胚胎成像与一种新的UPEFM，2）检测的 通过NTEFM使用反义技术的X连锁G6 PDH信息，3） 应用免疫组织化学技术检测发育中的桑椹胚和囊胚上的H-Y抗原 MPEFM，以及4）将G6 PDH胞浆水平和H-Y结合的成功 抗原检测以可靠地预测遗传性别。 的信息 在这些实验中获得的结果将增加我们的基本 利用实时显微镜了解胚胎发育。 的 开发一种检测方法，以正确预测遗传学上的性别， 在移植前操作胚胎将增加 这些进步是实现的。 此外，这项研究将 直接应用于提高繁殖效率， 国内物种

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.