Knowles Michelle K, Guenza Marina G, Capaldi Roderick A, Marcus Andrew H
Department of Chemistry and Materials Science Institute, University of Oregon, Eugene 97403, USA.
Proc Natl Acad Sci U S A. 2002 Nov 12;99(23):14772-7. doi: 10.1073/pnas.232346999. Epub 2002 Nov 4.
Subcellular organelle dynamics are strongly influenced by interactions with cytoskeletal filaments and their associated motor proteins, and lead to complex multiexponential relaxations that occur over a wide range of spatial and temporal scales. Here we report spatio-temporal measurements of the fluctuations of the mitochondrial reticulum in osteosarcoma cells by using Fourier imaging correlation spectroscopy, over time and distance scales of 10(-2) to 10(3) s and 0.5-2.5 microm. We show that the method allows a more complete description of mitochondrial dynamics, through the time- and length-scale-dependent collective diffusion coefficient D(k,tau), than available by other means. Addition of either nocodazole to disrupt microtubules or cytochalasin D to disassemble microfilaments simplifies the intermediate scattering function. When both drugs are used, the reticulum morphology of mitochondria is retained even though the cytoskeletal elements have been de-polymerized. The dynamics of the organelle are then primarily diffusive and can be modeled as a collection of friction points interconnected by elastic springs. This study quantitatively characterizes organelle dynamics in terms of collective cytoskeletal interactions in living cells.
亚细胞器动力学受到与细胞骨架细丝及其相关运动蛋白相互作用的强烈影响,并导致在广泛的空间和时间尺度上发生复杂的多指数弛豫。在这里,我们报告了通过使用傅里叶成像相关光谱法,在10^(-2)到10^(3)秒以及0.5 - 2.5微米的时间和距离尺度上,对骨肉瘤细胞中线粒体网状结构波动的时空测量。我们表明,与其他方法相比,该方法通过与时间和长度尺度相关的集体扩散系数D(k,τ),能够更完整地描述线粒体动力学。添加诺考达唑以破坏微管或细胞松弛素D以拆解微丝会简化中间散射函数。当同时使用这两种药物时,即使细胞骨架成分已经解聚,线粒体的网状形态仍得以保留。然后,细胞器的动力学主要是扩散性的,并且可以被建模为通过弹性弹簧相互连接的摩擦点的集合。这项研究从活细胞中集体细胞骨架相互作用的角度定量表征了细胞器动力学。