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Abstract：
The nervous system may include more than 100 residue-specific posttranslational modifications of histones forming the nucleosome core that are often regulated in cell-type-specific manner. On a genome-wide scale, some of the histone posttranslational modification landscapes show significant overlap with the genetic risk architecture for several psychiatric disorders, fueling PsychENCODE and other large-scale efforts to comprehensively map neuronal and nonneuronal epigenomes in hundreds of specimens. However, practical guidelines for efficient generation of histone chromatin immunoprecipitation followed by deep sequencing (ChIP-seq) datasets from postmortem brains are needed.
Protocols and quality controls are given for the following: 1) extraction, purification, and NeuN neuronal marker immunotagging of nuclei from adult human cerebral cortex; 2) fluorescence-activated nuclei sorting; 3) preparation of chromatin by micrococcal nuclease digest; 4) ChIP for open chromatin-associated histone methylation and acetylation; and 5) generation and sequencing of ChIP-seq libraries.
We present a ChIP-seq pipeline for epigenome mapping in the neuronal and nonneuronal nuclei from the postmortem brain. This includes a stepwise system of quality controls and user-friendly data presentation platforms.
Our practical guidelines will be useful for projects aimed at histone posttranslational modification mapping in chromatin extracted from hundreds of postmortem brain samples in cell-type-specific manner.
Protocols and quality controls are given for the following: 1) extraction, purification, and NeuN neuronal marker immunotagging of nuclei from adult human cerebral cortex; 2) fluorescence-activated nuclei sorting; 3) preparation of chromatin by micrococcal nuclease digest; 4) ChIP for open chromatin-associated histone methylation and acetylation; and 5) generation and sequencing of ChIP-seq libraries.
We present a ChIP-seq pipeline for epigenome mapping in the neuronal and nonneuronal nuclei from the postmortem brain. This includes a stepwise system of quality controls and user-friendly data presentation platforms.
Our practical guidelines will be useful for projects aimed at histone posttranslational modification mapping in chromatin extracted from hundreds of postmortem brain samples in cell-type-specific manner.
摘要：
神经系统可以包括形成核小体核心的组蛋白的超过100个残基特异性翻译后修饰,其通常以细胞类型特异性方式被调节。在全基因组范围内，一些组蛋白翻译后修饰景观显示与几种精神疾病的遗传风险结构显著重叠，推动PsychENCODE和其他大规模的努力，以全面绘制数百个标本中的神经元和非神经元表观基因组。然而,需要有效生成组蛋白染色质免疫沉淀,然后从死后大脑进行深度测序(ChIP-seq)数据集的实用指南.
给出了以下方案和质量控制：1)提取，净化,和NeuN神经元标记对成年人大脑皮层的核进行免疫标记；2）荧光激活的核分选；3）通过微球菌核酸酶消化制备染色质；4）ChIP用于开放染色质相关组蛋白甲基化和乙酰化；5）ChIP-seq文库的生成和测序。
我们提出了一个ChIP-seq管道，用于死后大脑的神经元和非神经元核中的表观基因组作图。这包括逐步的质量控制系统和用户友好的数据呈现平台。
我们的实用指南对于以细胞类型特异性方式从数百个死后大脑样本中提取的染色质中的组蛋白翻译后修饰作图的项目将是有用的。
给出了以下方案和质量控制：1)提取，净化,和NeuN神经元标记对成年人大脑皮层的核进行免疫标记；2）荧光激活的核分选；3）通过微球菌核酸酶消化制备染色质；4）ChIP用于开放染色质相关组蛋白甲基化和乙酰化；5）ChIP-seq文库的生成和测序。
我们提出了一个ChIP-seq管道，用于死后大脑的神经元和非神经元核中的表观基因组作图。这包括逐步的质量控制系统和用户友好的数据呈现平台。
我们的实用指南对于以细胞类型特异性方式从数百个死后大脑样本中提取的染色质中的组蛋白翻译后修饰作图的项目将是有用的。
