基于拉曼光谱的牛ICSI和IVF早期胚胎差异研究

项目名称： 基于拉曼光谱的牛ICSI和IVF早期胚胎差异研究

项目编号： No.31201801

项目类型： 青年科学基金项目

立项/批准年度： 2013

项目学科： 畜牧学与草地科学、兽医学、水产学

项目作者： 李晓霞

作者单位： 河南科技大学

项目金额： 22万元

中文摘要： 在辅助生殖领域，卵母细胞胞质内显微受精技术是研究受精和早期胚胎发育机制的一种重要方法；然而，与IVF相比，由ICSI形成的受精卵经体外培养后囊胚率、出生率都明显降低，这种低效率严重限制其被更广泛地应用。本课题组的前期研究表明激光光镊拉曼光谱技术可作为一种快速、有效且无损伤地评估精子品质的有力工具；我们推测，此技术可能成为研究早期胚胎发育的一种更精确、有效的方法。据此，本研究拟采用激光光镊拉曼光谱技术结合荧光标记法对比分析牛IVF和ICSI早期胚胎代谢组学、线粒体分布和细胞凋亡的差异；利用激光光镊拉曼光谱仪分析不同发育阶段的牛早期胚胎及其培养液的结构和组份变化，通过数学模型分析和对特征拉曼光谱的归属分析，探索ICSI胚胎发育率低的原因，从而为提高ICSI的效率和成功率奠定理论和方法基础，也为单个细胞水平上研究其结构和组分差异提供一个全新的视角。

中文关键词： 胞质内显微受精；早期胚胎；拉曼光谱；活性线粒体；细胞凋亡

英文摘要： Intracytoplasmic sperm injection (ICSI) is a powerful technique in the ?eld of assisted reproduction (ART) and provides exciting opportunities for studying the basic mechanisms of fertilization and early embryo development. Nevertheless, its application in agriculture and conservation biology has been greatly hampered by the low success rate reported for this method in respect of economically important species. Specifically, the rates of blastocyst formation and live newborn are greatly reduced when zygotes are generated by ICSI. ICSI remains a low e?ciency technology in comparison with alternatives such as in vitro fertilization (IVF) and its application is less widespread. Our previous study imply the potential of LTRS technique as a rapid, effective and noninvasive tool in assessing the quality of buffalo sperm cells. This technology can offer a most effective and precision method to allow rapid noninvasive assessment of embryonic reproductive potential before transfer. In this study, we determine differences in metabonomics, translocation of active mitochondria and cell apoptosis of early embryos generated by ICSI versus the less manipulative IVF with Laser Tweezers Raman Spectroscopy (LTRS) and fluorescence-labeled technique, providing dynamic change regularity. LTRS is used to investigate bovine early embr

英文关键词： ICSI；early embryos；Raman spectroscopy；active mitochondria；cell apoptosis