Live CLEM imaging of Tetrahymena to analyze the dynamic behavior of the nuclear pore complex
Live CLEM imaging of Tetrahymena to analyze the dynamic behavior of the nuclear pore complex
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四膜虫实时 CLEM 成像，分析核孔复合体的动态行为
DOI：
10.1007/978-1-0716-2337-4_30
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发表时间：
2022
影响因子：
--
通讯作者：
Masaaki Iwamoto
中科院分区：
文献类型：
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作者：
Tokuko Haraguchi;Hiroko Osakada;Masaaki Iwamoto
四膜虫是研究核孔复合体的理想生物，因为它在细胞内有两个结构和功能不同的核(大核和微核)，每个核中有两个成分不同、功能不同的核孔复合体(NPC)。因此，有可能将特定组成蛋白的功能与大核和微核的核功能联系起来。此外，在核分化过程中，这些NPC的结构和组成也会发生动态变化。LIVE CLEM成像是相关光学和电子显微镜(CLEM)与活细胞成像相结合的一种方法，是以高分辨率可视化特定感兴趣分子/结构的动态变化的有力工具。在这里，我们描述了可以应用于研究四膜细胞在核分化过程中神经前体的动态行为的Live Clem。
Tetrahymenais a fascinating organism for studying the nuclear pore complex because it has two structurally and functionally distinct nuclei (macronucleus and micronucleus) within a cell, and there are two compositionally distinct nuclear pore complexes (NPCs) with different functions in each nucleus. Therefore, it is possible to link the function of a specific constituent protein with the nuclear function of the macronucleus and micronucleus. Additionally, these NPCs undergo dynamic changes in their structures and compositions during nuclear differentiation. Live CLEMimaging, a method of correlative light and electron microscopy (CLEM) combined with live cell imaging, is a powerful tool for visualizing these dynamic changes of specific molecules/structures of interest at high resolution. Here, we describe Live CLEM that can be applied to the study of the dynamic behavior of NPCs inTetrahymenacells undergoing nuclear differentiation.