A high-resolution imaging approach to investigate chromatin architecture in complex tissues.
A high-resolution imaging approach to investigate chromatin architecture in complex tissues.
复制标题
DOI：
10.1016/j.cell.2015.09.002
复制
发表时间：
2015-09-24
期刊：
影响因子：
64.5
通讯作者：
Mandel G
中科院分区：
文献类型：
--
作者：
Linhoff MW;Garg SK;Mandel G
我们提出了 ChromATin，一种用于研究复杂组织中染色质组织的定量高分辨率成像方法。该方法结合了通过免疫染色对表观遗传修饰进行分析、通过 FISH 对特定 DNA 序列进行定位，以及使用阵列断层扫描 (AT) 成像对核区室进行高分辨率分离。然后，我们应用这种方法来了解雌性雷特综合症小鼠的哺乳动物大脑中的基因组是如何组织的，这些小鼠是正常细胞和 Mecp2 缺失细胞的镶嵌体。同一区域内的并排比较揭示了野生型神经元中不同的异染色质区域，这些区域在 Mecp2-null 细胞核中发生了改变。突变神经元表现出染色质压缩增加，并且 H4K20me3 组蛋白修饰显着重新分布到着丝粒周围异染色质（通常由 MeCP2 占据的区域）。这些事件并非在每种神经元细胞类型中都观察到，这凸显了 ChromATin 作为一种强大的原位方法，可用于检查复杂组织中细胞类型特异性染色质结构的差异。
We present ChromATin, a quantitative high-resolution imaging approach for investigating chromatin organization in complex tissues. This method combines analysis of epigenetic modifications by immunostaining, localization of specific DNA sequences by FISH, and high-resolution segregation of nuclear compartments using array tomography (AT) imaging. We then apply this approach to understand how the genome is organized in the mammalian brain using female Rett Syndrome mice, which are a mosaic of normal and Mecp2-null cells. Side-by-side comparisons within the same field reveal distinct heterochromatin territories in wild type neurons that are altered in Mecp2-null nuclei. Mutant neurons exhibit increased chromatin compaction and a striking redistribution of the H4K20me3 histone modification into pericentromeric heterochromatin, a territory occupied normally by MeCP2. These events are not observed in every neuronal cell type, highlighting ChromATin as a powerful in situ method for examining cell type-specific differences in chromatin architecture in complex tissues.