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Cell:利用荧光追踪细胞周期的新技术

来源:Cell 2008-02-14 11:47

(封面图片:处于不同细胞周期的细胞核发出红色或绿色荧光)

对于很多生物学领域的研究而言,分析细胞周期变化过程中的分子和细胞学变化都是非常重要的,特别是G1-S期的变化,然而这种变化非常难以观察。在2008年2月8日出版的《细胞》(Cell)文章中来自日本的一组科学家表示,他们得到了两种新型荧光蛋白,其中一种能使得G1期细胞核呈现红色,而另一种使得S/G2/M期细胞核呈现出绿色。

蛋白质“Cdtl”只存在于完成分裂的细胞准备复制DNA的G1期,蛋白质“Geminin”只存在于G1期之外的细胞周期其他阶段。研究人员向两种蛋白质分别植入红色和绿色荧光物质的荧光基因,将处于G1期的细胞核标记为红色,处于其他时期的细胞核标记为绿色。

在研究中,科学家将红色荧光蛋白以及绿色荧光蛋白结合到E3酶底物、Cdt1和蛋白Geminin上,从而得到了双色荧光探针,来分辨活细胞处于细胞周期的哪一阶段。结果发现,表达这些探针的细胞以及转基因小鼠的每一个细胞核都发出红色或绿色的荧光。研究人员将这些蛋白称为荧光泛素化细胞周期标志物(Fucci)。此外,研究小组还使用延时成像技术研究了细胞在上皮-间质转分化过程中发生的细胞周期动态变化,结果表明,新型荧光探针能较好的用于分辨活细胞周期的时间和空间变化。

荧光探针Fucci还能用于分析大脑中神经祖细胞的分化和转移,以及活体小鼠血管肿瘤的发生等等。这些小鼠和细胞结果能作为模型系统,帮助科学家们更好的了解细胞周期在多种生物活动中的情况。Fucci技术能实现双色成像,并且具有很好的亮度和对比度。科学家们表示,Fucci技术接下来面临的主要挑战是如何发展Fucci衍生物,使得衍生物拥有其它颜色,从而用来区分细胞周期中除了G1和S之外的其它阶段。(科学网 何宏辉/编译)

(《细胞》(Cell),Vol 132, 487-498, 08 February 2008,Asako Sakaue-Sawano, Atsushi Miyawaki)

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Cell, Vol 132, 487-498, 08 February 2008

Visualizing Spatiotemporal Dynamics of Multicellular Cell-Cycle Progression

Asako Sakaue-Sawano,1,3 Hiroshi Kurokawa,1,4 Toshifumi Morimura,2 Aki Hanyu,5 Hiroshi Hama,1 Hatsuki Osawa,1 Saori Kashiwagi,2 Kiyoko Fukami,4 Takaki Miyata,6 Hiroyuki Miyoshi,7 Takeshi Imamura,5 Masaharu Ogawa,2 Hisao Masai,8 and Atsushi Miyawaki1,3,

1 Laboratory for Cell Function and Dynamics, Advanced Technology Development Group, Brain Science Institute, RIKEN, 2-1 Hirosawa, Wako-city, Saitama 351-0198, Japan
2 Laboratory for Cell Culture Development, Advanced Technology Development Group, Brain Science Institute, RIKEN, 2-1 Hirosawa, Wako-city, Saitama 351-0198, Japan
3 Life Function and Dynamics, ERATO, JST, 2-1 Hirosawa, Wako-city, Saitama 351-0198, Japan
4 School of Life Science, Tokyo University of Pharmacy and Life Science, 1432-1 Horinouchi, Hachioji, Tokyo 192-0392, Japan
5 Departments of Biochemistry, The Cancer Institute of the Japanese Foundation for Cancer Research, 3-10-6 Ariake, Koto-ku, Tokyo 135-8550, Japan
6 Department of Anatomy and Cell Biology, Nagoya University Graduate School of Medicine, 65 Tsurumai-cho, Syowa-ku, Nagoya, Aichi 466-8550, Japan
7 Subteam for Manipulation of Cell Fate, BioResource Center, RIKEN Tsukuba Institute, 3-1-1 Koyadai, Tsukuba, Ibaraki 305-0074, Japan
8 Genome Dynamics Project, Tokyo Metropolitan Institute of Medical Science, 3-18-22 Honkomagome, Bunkyo-ku, Tokyo 113-8613, Japan

Corresponding author
Atsushi Miyawaki
matsushi@brain.riken.jp

Summary

The cell-cycle transition from G1 to S phase has been difficult to visualize. We have harnessed antiphase oscillating proteins that mark cell-cycle transitions in order to develop genetically encoded fluorescent probes for this purpose. These probes effectively label individual G1 phase nuclei red and those in S/G2/M phases green. We were able to generate cultured cells and transgenic mice constitutively expressing the cell-cycle probes, in which every cell nucleus exhibits either red or green fluorescence. We performed time-lapse imaging to explore the spatiotemporal patterns of cell-cycle dynamics during the epithelial-mesenchymal transition of cultured cells, the migration and differentiation of neural progenitors in brain slices, and the development of tumors across blood vessels in live mice. These mice and cell lines will serve as model systems permitting unprecedented spatial and temporal resolution to help us better understand how the cell cycle is coordinated with various biological events.

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