4-D single particle tracking of synthetic and proteinaceous microspheres reveals preferential movement of nuclear particles along chromatin - poor tracks.
4-D single particle tracking of synthetic and proteinaceous microspheres reveals preferential movement of nuclear particles along chromatin - poor tracks.
复制标题
DOI：
10.1186/1471-2121-5-45
复制
发表时间：
2004-11-23
期刊：
影响因子：
--
通讯作者：
Eils R
中科院分区：
文献类型：
--
作者：
Bacher CP;Reichenzeller M;Athale C;Herrmann H;Eils R
核组织、核体，特别是RNPs的动力学一直是许多研究的焦点。要了解它们的功能，了解它们的空间核位置和时间易位是必不可少的。通常，这样的研究产生了大量的数据，需要图像分析和计算工具中的新方法来定量跟踪移动细胞和形状变化的核背景上的粒子运动。我们开发了一种新型的4-D图像处理平台(Tikal)，用于激光扫描和广域显微镜的工作。Tikal提供了一个注册软件，用于纠正细胞的全局运动和局部变形，以及2-D和3-D跟踪软件。利用这一新工具，我们用活细胞时间推移显微镜结合单粒子跟踪和迁移率分析，研究了两种不同类型的核粒子，即由绿色荧光蛋白-NLS-波形蛋白制成的核体和显微注射0.1MPs m宽的聚苯乙烯微珠的动力学。我们现在提供了一种工具，用于在获取染色质密度数据的同时对颗粒运动进行自动3-D分析。动力学分析显示4种运动方式：受限阻塞运动、正常扩散运动和定向运动。对染色质背景的粒子跟踪表明，粒子的运动与染色质的局部重组直接相关。此外，对核质和细胞质中颗粒运动的直接比较表明，波形蛋白颗粒在两个隔室中的动力学行为完全不同。核粒子的动力学受ATP耗竭的影响较小，而受到肌动蛋白和微管网络破坏的显著干扰。此外，细胞核的水化状态对核体的运动有很大的影响，因为当细胞受到0.6M山梨醇的攻击时，正常的扩散和定向运动都被完全取消。这一效应与染色质的紧密结合有关。我们得出结论，染色质密度的变化直接影响核内蛋白质组装的流动性。
The dynamics of nuclear organization, nuclear bodies and RNPs in particular has been the focus of many studies. To understand their function, knowledge of their spatial nuclear position and temporal translocation is essential. Typically, such studies generate a wealth of data that require novel methods in image analysis and computational tools to quantitatively track particle movement on the background of moving cells and shape changing nuclei. We developed a novel 4-D image processing platform (TIKAL) for the work with laser scanning and wide field microscopes. TIKAL provides a registration software for correcting global movements and local deformations of cells as well as 2-D and 3-D tracking software. With this new tool, we studied the dynamics of two different types of nuclear particles, namely nuclear bodies made from GFP-NLS-vimentin and microinjected 0.1 μm – wide polystyrene beads, by live cell time-lapse microscopy combined with single particle tracking and mobility analysis. We now provide a tool for the automatic 3-D analysis of particle movement in parallel with the acquisition of chromatin density data. Kinetic analysis revealed 4 modes of movement: confined obstructed, normal diffusion and directed motion. Particle tracking on the background of stained chromatin revealed that particle movement is directly related to local reorganization of chromatin. Further a direct comparison of particle movement in the nucleoplasm and the cytoplasm exhibited an entirely different kinetic behaviour of vimentin particles in both compartments. The kinetics of nuclear particles were slightly affected by depletion of ATP and significantly disturbed by disruption of actin and microtubule networks. Moreover, the hydration state of the nucleus had a strong impact on the mobility of nuclear bodies since both normal diffusion and directed motion were entirely abolished when cells were challenged with 0.6 M sorbitol. This effect correlated with the compaction of chromatin. We conclude that alteration in chromatin density directly influences the mobility of protein assemblies within the nucleus.
登录
查看更多内容
DOI：
10.1083/jcb.200302097
发表时间：
2003-05-12
期刊：
The Journal of cell biology
影响因子：
--
作者：
Eils R;Athale C
通讯作者：
Athale C
影响因子：
3
作者：
Herrmann, H;Wedig, T;Aebi, U
通讯作者：
Aebi, U
影响因子：
3.4
作者：
Apgar, J;Tseng, Y;Wirtz, D
通讯作者：
Wirtz, D
影响因子：
3.7
作者：
HEDBERG, KK;CHEN, LB
通讯作者：
CHEN, LB
影响因子：
56.9
作者：
Heun, P;Laroche, T;Gasser, SM
通讯作者：
Gasser, SM