作者：Gill; SE; Halene; T; Rajarajan; P; Cha...chromosomeconformationtn5transposasedopaminergicneuronsubstantianigra
摘要：Objective:Monoamine system plays a key role in the pathophysiology of a wide range of neuropsychiatric disorders including Parkinson’s,Huntington’s,drug addiction and many other conditions.Of note,there is a large amount of literatures exploring monoamine pathways in the context of adaptive mechanisms inside the cell nucleus,including changes in gene expression and alterations in chromatin modifications,structure and functions.However,such types of studies were largely focused on forebrain structures that receive input from monoaminergic neurons.Comprehensive genome-scale and cell-type specific mappings is lacking for brainstem areas that consist of monoaminergic neurons,including the substantia nigra(SN)and ventral tegmental area(VTA).This is especially the case for human study,due to the limited access to clinical human brain samples and lack of efficient assay for cell-type specific isolation from postmortem brain tissues after low-temperature or fixative preservations.Here,we introduce novel protocol to comprehensively map the 3D genome and nuclear transcriptome in selective population of dopaminergic neurons isolated from human midbrain.Methods:Mammalian chromatin is highly packed and compartmentalized into Topologically Associated Domains(TADs).Although TAD structures were reported to conserve in between species and cell types,chromatin contacts within TAD were often highly specific for different cell types in brain,and were implicated in the genetic risk of psychiatric disorders.Fragmentationreligation based DNA-DNA proximity(Hi-C)assays were designed to detect chromosomal conformations,which provide an additional layer of epigenomic regulation on top of previous well-studied epigenetic mechanisms including DNA and histone modifications.However,current Hi-C protocols either require millions of cells as input,or only offer limited resolution when applied to low input material,particularly when probing rare cell types from human postmortem brain tissue.Here,we designed Tn5-Hi-C,a modified Hi-C protocol t
注：因版权方要求，不能公开全文，如需全文，请咨询杂志社
