Probing the Stochastic, Motor-Driven Properties of the Cytoplasm Using Force Spectrum Microscopy

被引:373
|
作者
Guo, Ming [1 ]
Ehrlicher, Allen J. [1 ,2 ]
Jensen, Mikkel H. [1 ,3 ]
Renz, Malte [4 ]
Moore, Jeffrey R. [3 ]
Goldman, Robert D. [5 ]
Lippincott-Schwartz, Jennifer [4 ]
Mackintosh, Frederick C. [6 ]
Weitz, David A. [1 ,7 ]
机构
[1] Harvard Univ, Sch Engn & Appl Sci, Cambridge, MA 02138 USA
[2] Beth Israel Deaconess Med Ctr, Boston, MA 02115 USA
[3] Boston Univ, Dept Phys & Biophys, Boston, MA 02118 USA
[4] Eunice Kennedy Shriver Natl Inst Child Hlth & Hum, NIH, Bethesda, MD 20892 USA
[5] Northwestern Univ, Feinberg Sch Med, Dept Cell & Mol Biol, Chicago, IL 60611 USA
[6] Vrije Univ Amsterdam, Dept Phys & Astron, NL-1081 HV Amsterdam, Netherlands
[7] Harvard Univ, Dept Phys, Cambridge, MA 02138 USA
关键词
BREAST-CANCER CELLS; LIVING CELLS; MOLECULAR MOTORS; CHROMOSOMAL LOCI; DIFFUSION; MECHANICS; DYNAMICS; MICRORHEOLOGY; SPECTROSCOPY; MIGRATION;
D O I
10.1016/j.cell.2014.06.051
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Molecular motors in cells typically produce highly directed motion; however, the aggregate, incoherent effect of all active processes also creates randomly fluctuating forces, which drive diffusive-like, non-thermal motion. Here, we introduce force-spectrum-microscopy (FSM) to directly quantify random forces within the cytoplasm of cells and thereby probe stochastic motor activity. This technique combines measurements of the random motion of probe particles with independent micromechanical measurements of the cytoplasm to quantify the spectrum of force fluctuations. Using FSM, we show that force fluctuations substantially enhance intracellular movement of small and large components. The fluctuations are three times larger in malignant cells than in their benign counterparts. We further demonstrate that vimentin acts globally to anchor organelles against randomly fluctuating forces in the cytoplasm, with no effect on their magnitude. Thus, FSM has broad applications for understanding the cytoplasm and its intracellular processes in relation to cell physiology in healthy and diseased states.
引用
收藏
页码:822 / 832
页数:11
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