A novel live-cell imaging system reveals a reversible hydrostatic pressure impact on cell-cycle progression.
A novel live-cell imaging system reveals a reversible hydrostatic pressure impact on cell-cycle progression.
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DOI：
10.1242/jcs.212167
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发表时间：
2018-08-06
影响因子：
4
通讯作者：
Geeves MA
中科院分区：
文献类型：
--
作者：
Brooker HR;Gyamfi IA;Wieckowska A;Brooks NJ;Mulvihill DP;Geeves MA
生命依赖于细胞对环境变化迅速作出反应的能力。局部环境中的微小扰动会改变分子相互作用的能力，从而改变分子之间的交流。静水压提供了一种快速、无创、完全可逆的方法来调节体内和体外分子之间的亲和力。我们开发了一种简单的荧光成像室，可以在活细胞中在压力<200 bar的情况下跟踪细胞内蛋白质动力学和分子事件。通过使用酵母，我们研究了静水压力对细胞生长和细胞周期进程的影响。虽然100巴对活力没有影响，但它诱导染色体分离延迟，导致长未分裂细胞的积累，这些细胞也是弯曲的，与细胞骨架的破坏一致。这种延迟是独立的压力信号，并诱导细胞周期进程的同步。在白色念珠菌中观察到了相同的效果，压力诱导可逆的细胞周期延迟和菌丝生长。我们提出了一种简单的新型非侵入性荧光显微镜为基础的方法，瞬时影响分子动力学，以可视化，解剖和研究活细胞中的信号通路和细胞过程。总结：开发一种简单的荧光成像室，可以在高达200 bar的压力下观察活细胞中的细胞内蛋白质动力学和分子事件。
Life is dependent upon the ability of a cell to rapidly respond to changes in the environment. Small perturbations in local environments change the ability of molecules to interact and, hence, communicate. Hydrostatic pressure provides a rapid non-invasive, fully reversible method for modulating affinities between molecules both in vivo and in vitro. We have developed a simple fluorescence imaging chamber that allows intracellular protein dynamics and molecular events to be followed at pressures <200 bar in living cells. By using yeast, we investigated the impact of hydrostatic pressure upon cell growth and cell-cycle progression. While 100 bar has no effect upon viability, it induces a delay in chromosome segregation, resulting in the accumulation of long undivided cells that are also bent, consistent with disruption of the cytoskeletons. This delay is independent of stress signalling and induces synchronisation of cell-cycle progression. Equivalent effects were observed in Candida albicans, with pressure inducing a reversible cell-cycle delay and hyphal growth. We present a simple novel non-invasive fluorescence microscopy-based approach to transiently impact molecular dynamics in order to visualise, dissect and study signalling pathways and cellular processes in living cells. Summary: Development of a simple fluorescence imaging chamber allowing observation of intracellular protein dynamics and molecular events in living cells at pressure up to 200 bar.
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影响因子：
3.3
作者：
George, Vinoj T.;Brooks, Gavin;Humphrey, Timothy C.
通讯作者：
Humphrey, Timothy C.
影响因子：
2.9
作者：
Pearson, David S.;Swartz, Darl R.;Geeves, Michael A.
通讯作者：
Geeves, Michael A.
影响因子：
2
作者：
CONTI, F;INOUE, I;STUHMER, W
通讯作者：
STUHMER, W
影响因子：
3.5
作者：
DAVIS, JS;GUTFREUND, H
通讯作者：
GUTFREUND, H
DOI：
10.1083/jcb.140.2.367
发表时间：
1998-01-26
期刊：
The Journal of cell biology
影响因子：
--
作者：
Roberts TM;Salmon ED;Stewart M
通讯作者：
Stewart M