The changing biochemical composition and organisation of the murine oocyte and early embryo as revealed by Raman spectroscopic mapping
The changing biochemical composition and organisation of the murine oocyte and early embryo as revealed by Raman spectroscopic mapping
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DOI：
10.1002/jrs.2964
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发表时间：
2012-01-01
影响因子：
2.5
通讯作者：
Elfick, Alistair
中科院分区：
文献类型：
--
作者：
Davidson, Bryony;Spears, Norah;Elfick, Alistair
尽管近年来辅助生殖技术（例如体外受精）的应用呈指数级增长，但人们对卵子和胚胎发育和成熟过程中发生的生化修饰仍不完全了解。因此，为了提高这些技术的效率，有必要进一步了解支持卵母细胞和胚胎发育的过程。拉曼光谱图作为一种技术，无需标记即可研究完整细胞内的生化变异。这里，使用 785 nm 激发和 2 μm 步长收集固定的未成熟和成熟卵母细胞以及早期胚胎的拉曼图。使用单变量和多变量技术对结果进行分析。研究发现，卵母细胞成熟过程中发生了显着的大分子积累，而在胚胎卵裂时观察到总脂质含量下降，这与新细胞膜的形成一致。此外，在成熟卵母细胞中观察到的大分子的不对称定位可能表明细胞质极化的存在，这种现象已在低等生物的卵中观察到。因此，这些结果表明拉曼光谱图可能为研究卵子和胚胎发育的生物化学提供替代分析工具。特别是，这些结果表明时间拉曼分析可能有助于揭示鼠卵中细胞质极化的存在。版权所有 (c) 2011 John Wiley & Sons, Ltd.
Despite an exponential uptake in recent years of assisted reproductive techniques, such as in vitro fertilisation, much is still not fully understood about the biochemical modifications that take place during the development and maturation of the egg and embryo. As such, in order to improve the efficiency of these techniques, furthering our understanding of the processes that underpin oocyte and embryo development is necessary. Raman spectroscopic mapping as a technique enables the investigation of biochemical variation within intact cells without the need for labelling. Here, Raman maps of fixed immature and mature oocytes along with early stage embryos were collected using 785 nm excitation and a step size of 2 mu m. The results were analysed using both univariate and multivariate techniques. It was found that significant macromolecular accumulation took place during oocyte maturation, while a decrease in total lipid content consistent with the formation of new cellular membranes is observed upon embryo cleavage. Furthermore, an observed asymmetrical localisation of macromolecules in the mature oocyte may indicate the existence of cytoplasmic polarisation, a phenomenon that has been observed in the eggs of lower organisms. As such, these results indicate that Raman spectroscopic mapping may present an alternative analytical tool for investigating the biochemistry of egg and embryo development. In particular, these results indicate that temporal Raman analysis may help to reveal the existence of cytoplasmic polarisation in the murine egg. Copyright (c) 2011 John Wiley & Sons, Ltd.