Direct visualization of cell division using high-resolution imaging of M-phase of the cell cycle.
Direct visualization of cell division using high-resolution imaging of M-phase of the cell cycle.
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DOI：
10.1038/ncomms2089
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发表时间：
2012
影响因子：
16.6
通讯作者：
中科院分区：
文献类型：
--
作者：
目前监测和量化活细胞中的细胞分裂以及可靠地区分细胞分裂停止和核内复制的方法是有限的，并且使干细胞库身份的确定复杂化。在这里，我们克服了这些限制，通过产生一个在体内报告系统使用的支架蛋白质苯胺融合增强绿色荧光蛋白，提供高时空分辨率的有丝分裂期。这种方法将胞质分裂和中间体形成可视化为干细胞和体细胞扩增的标志，并且能够与细胞周期变化区分开。高分辨率显微镜在胚胎心脏和脑组织中的增强型绿色荧光蛋白-苯胺醛转基因小鼠允许实时监测细胞分裂和定量细胞周期动力学。对损伤后心脏细胞分裂的分析表明，梗死边缘区心肌细胞的倍性增加，但细胞分裂不增加。因此，增强的绿色荧光蛋白-苯胺醛系统能够在体内监测和测量细胞分裂，并显著简化固定细胞的体外分析。目前用于检测活细胞增殖的方法不能区分分裂细胞和通过细胞周期进展的细胞。在这里，描述了一种方法，该方法在M期期间检测细胞的收缩环和中间体中的苯胺，从而提供分裂细胞的更准确的检测。
Current approaches to monitor and quantify cell division in live cells, and reliably distinguish between acytokinesis and endoreduplication, are limited and complicate determination of stem cell pool identities. Here we overcome these limitations by generating an in vivo reporter system using the scaffolding protein anillin fused to enhanced green fluorescent protein, to provide high spatiotemporal resolution of mitotic phase. This approach visualizes cytokinesis and midbody formation as hallmarks of expansion of stem and somatic cells, and enables distinction from cell cycle variations. High-resolution microscopy in embryonic heart and brain tissues of enhanced green fluorescent protein–anillin transgenic mice allows live monitoring of cell division and quantitation of cell cycle kinetics. Analysis of cell division in hearts post injury shows that border zone cardiomyocytes in the infarct respond with increasing ploidy, but not cell division. Thus, the enhanced green fluorescent protein–anillin system enables monitoring and measurement of cell division in vivo and markedly simplifies in vitro analysis in fixed cells. Current methods for detecting proliferation in live cells cannot distinguish between dividing cells and cells that are progressing through the cell cycle. Here, a method is described that detects anillin in the contractile ring and in the midbody of cells during M-phase, providing a more accurate detection of dividing cells.
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影响因子：
3.7
作者：
Haberlandt C;Derouiche A;Wyczynski A;Haseleu J;Pohle J;Karram K;Trotter J;Seifert G;Frotscher M;Steinhäuser C;Jabs R
通讯作者：
Jabs R
影响因子：
10.8
作者：
Meckert, PC;Rivello, HG;Laguens, R
通讯作者：
Laguens, R
影响因子：
10.8
作者：
Herget, GW;Neuburger, M;Adler, CP
通讯作者：
Adler, CP
DOI：
10.1073/pnas.0903089106
发表时间：
2009-10-06
影响因子：
11.1
作者：
Hosoda, Toru;D'Amario, Domenico;Leri, Annarosa
通讯作者：
Leri, Annarosa
影响因子：
5.6
作者：
Becker, Klaus A.;Ghule, Prachi N.;Stein, Gary S.
通讯作者：
Stein, Gary S.