New quantitative approaches reveal the spatial preference of nuclear compartments in mammalian fibroblasts.
New quantitative approaches reveal the spatial preference of nuclear compartments in mammalian fibroblasts.
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DOI：
10.1098/rsif.2014.0894
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发表时间：
2015-03-06
期刊：
影响因子：
--
通讯作者：
Freemont PS
中科院分区：
文献类型：
--
作者：
Weston DJ;Russell RA;Batty E;Jensen K;Stephens DA;Adams NM;Freemont PS
高等真核细胞的细胞核显示出区室化，并且某些核区室已被证明遵循一定程度的空间组织。迄今为止，对核组织的研究往往涉及简单的定量程序，这些程序既要解决核边界的不规则性，又要解决处理重复图像的问题。这些研究通常集中在物体间的距离，而不是在核内的空间位置。本文关注的是核室的空间偏好，为此我们开发了统计工具来定量研究和探索核组织。这些工具将联合收割机复制的图像结合起来，生成代表核室空间偏好的“聚合图”。我们提出了两个例子，不同的隔间在哺乳动物成纤维细胞（WI-38和MRC-5），展示了新的知识，细胞核内的空间偏好。具体而言，RNA聚合酶II的空间偏好在正常细胞和永生化细胞中得以保留，而PML核体表现出空间偏好的变化，从避免正常细胞中的中心到表现出对永生化细胞中的中心的偏好。此外，我们发现，SC 35剪接斑点被排除在核边界和本地化的整个核质和非转化WI-38细胞的染色质间空间。因此，这种新的方法能够揭示大规模扰动对空间结构和偏好的影响，这在单细胞成像中并不明显。
The nuclei of higher eukaryotic cells display compartmentalization and certain nuclear compartments have been shown to follow a degree of spatial organization. To date, the study of nuclear organization has often involved simple quantitative procedures that struggle with both the irregularity of the nuclear boundary and the problem of handling replicate images. Such studies typically focus on inter-object distance, rather than spatial location within the nucleus. The concern of this paper is the spatial preference of nuclear compartments, for which we have developed statistical tools to quantitatively study and explore nuclear organization. These tools combine replicate images to generate ‘aggregate maps' which represent the spatial preferences of nuclear compartments. We present two examples of different compartments in mammalian fibroblasts (WI-38 and MRC-5) that demonstrate new knowledge of spatial preference within the cell nucleus. Specifically, the spatial preference of RNA polymerase II is preserved across normal and immortalized cells, whereas PML nuclear bodies exhibit a change in spatial preference from avoiding the centre in normal cells to exhibiting a preference for the centre in immortalized cells. In addition, we show that SC35 splicing speckles are excluded from the nuclear boundary and localize throughout the nucleoplasm and in the interchromatin space in non-transformed WI-38 cells. This new methodology is thus able to reveal the effect of large-scale perturbation on spatial architecture and preferences that would not be obvious from single cell imaging.
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DOI：
10.1038/nrg3454
发表时间：
2013-06
期刊：
Nature reviews. Genetics
影响因子：
--
作者：
通讯作者：
--
影响因子：
56.9
作者：
Kaiser, Trish E.;Intine, Robert V.;Dundr, Miroslav
通讯作者：
Dundr, Miroslav
影响因子：
3.7
作者：
Niedojadlo, Janusz;Perret-Vivancos, Cecile;Fakan, Stanislav
通讯作者：
Fakan, Stanislav
影响因子：
5.6
作者：
SISKEN, JE;BONNER, SV;GRASCH, SD
通讯作者：
GRASCH, SD
影响因子：
4
作者：
Nunez, Esperanza;Fu, Xiang-Dong;Rosenfeld, Michael G.
通讯作者：
Rosenfeld, Michael G.