Raman micro-spectroscopy can be used to investigate the developmental stage of the mouse oocyte.
Raman micro-spectroscopy can be used to investigate the developmental stage of the mouse oocyte.
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DOI：
10.1371/journal.pone.0067972
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发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Spears N
中科院分区：
文献类型：
--
作者：
Davidson B;Murray AA;Elfick A;Spears N
近年来，体外受精等辅助生殖技术的使用呈指数级增长。然而，关于卵母细胞发育和成熟过程中发生的生物化学修饰还有很多尚未完全了解。因此，有必要进一步了解这些程序中卵母细胞操作最终如何影响其发育潜力;然而，目前几乎没有能够提供卵母细胞质量定量测量的方法。拉曼光谱使得能够在不需要标记的情况下研究完整细胞的整体生化特征。在这里，拉曼光谱获得的卵质的小鼠卵母细胞在不同的发展阶段，从晚期前腔卵泡，收集后，在体外成熟的卵巢卵泡和刺激和刺激排卵周期。利用单变量和多变量统计方法相结合，发现卵浆脂质含量可以用来区分卵母细胞发育的不同阶段。此外，成熟卵母细胞的光谱曲线显示，与体内生长的卵母细胞相比，体外发育的卵母细胞是蛋白质缺乏的。最后，两个拉曼峰强度之比，即1605∶1447 cm−1，被用作卵质蛋白质与脂质比的代表，被证明是卵母细胞质量的指示。总之，结果表明，拉曼光谱可能会提出一个替代的分析工具，用于调查卵母细胞发育阶段和质量的生物化学。
In recent years, the uptake of assisted reproductive techniques such as in vitro fertilisation has risen exponentially. However, there is much that is still not fully understood about the biochemical modifications that take place during the development and maturation of the oocyte. As such, it is essential to further the understanding of how oocyte manipulation during these procedures ultimately affects its developmental potential; yet, there are few methods currently available which are capable of providing a quantitative measure of oocyte quality. Raman spectroscopy enables investigation of the global biochemical profile of intact cells without the need for labelling. Here, Raman spectra were acquired from the ooplasm of mouse oocytes at various stages of development, from late pre-antral follicles, collected after in vitro maturation within their ovarian follicles and from unstimulated and stimulated ovulatory cycles. Using a combination of univariate and multivariate statistical methods, it was found that ooplasm lipid content could be used to discriminate between different stages of oocyte development. Furthermore, the spectral profiles of mature oocytes revealed that oocytes which have developed in vitro are protein-deficient when compared to in vivo grown oocytes. Finally, the ratio of two Raman peak intensities, namely 1605∶1447 cm−1, used as a proxy for the protein-to-lipid ratio of the ooplasm, was shown to be indicative of the oocyte’s quality. Together, results indicate that Raman spectroscopy may present an alternative analytical tool for investigating the biochemistry of oocyte developmental stage and quality.
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