Novel morphological approaches for the study of oocyte maturation.
Novel morphological approaches for the study of oocyte maturation.
复制标题
研究卵母细胞成熟的新形态学方法。
DOI：
10.1095/biolreprod30.1.13
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发表时间：
1984
影响因子：
3.6
通讯作者：
Albertini,DF
中科院分区：
文献类型：
--
作者：
Albertini,DF
讨论了光学显微镜在研究哺乳动物卵母细胞减数分裂成熟过程中发生的细胞表面、细胞骨架和细胞器变化方面的应用的最新进展。考虑三种技术：1）使用免疫细胞化学或药理学探针进行多重荧光染料标记，以分析固定在减数分裂各个阶段的卵母细胞中成分的空间和时间分布，2）使用活体荧光染色剂来研究活卵母细胞中细胞器的实际运动，以及3）活细胞的视频图像增强显微镜，以增强荧光、偏振和微分干涉显微镜的光学能力来记录动态细胞变化。描述了一种分别使用 Hoechst 33258、抗微管蛋白抗体和 NBD-鬼笔环素同时定位固定啮齿动物卵母细胞中的染色体、微管和肌动蛋白的方法。使用吖啶橙和激光染料罗丹明 123 作为活体染色剂，分别观察正在经历减数分裂成熟的大鼠卵母细胞中的溶酶体和线粒体。最后，讨论了延时视频图像增强显微镜在荧光标记细胞成分研究中的应用，特别是对细胞器的扩展监测，以分析成熟过程中卵母细胞成分的动态运动。
Recent advances in light microscopy are discussed with respect to their application for the study of cell surface, cytoskeletal and organellar changes that occur during meiotic maturation in mammalian oocytes. Three techniques are considered:1) multiple fluorochrome labeling using immunocytochemical or pharmacological probes to analyze the spatial and temporal disposition of components in oocytes fixed at various stages of meiosis,2) the use of vital fluorescence stains to study the actual movement of organelles in living oocytes, and3) video image intensification microscopy of living cells to record dynamic cellular changes with enhanced optical capabilities for fluorescence, polarization and differential interference contrast microscopy. A method is described for simultaneously localizing chromosomes, microtubules and f-actin in fixed rodent oocytes using, respectively, Hoechst 33258, antitubulin antibodies and NBD-phallicidin. Acridine orange and the laser dye rhodamine 123 are employed as vital stains to visualize lysosomes and mitochondria, respectively, in rat oocytes undergoing meiotic maturation. Finally, the application of time-lapse video image intensification microscopy for the study of fluorescently labeled cellular components is discussed with special reference to extended monitoring of cellular organelles for the analysis of dynamic movements of oocyte constituents during maturation.