期刊
GENES
卷 13, 期 2, 页码 -出版社
MDPI
DOI: 10.3390/genes13020306
关键词
DNA methylation; methylome; DRD2; medium spiny neuron; nucleus accumbens; mouse
资金
- National Institutes of Health [DP1 DA046587, R01 DA046720]
- FSU Legacy Fellowship
DNA methylation plays important roles in cellular processes, but traditional methods have limitations in studying specific cell types. This study applied two new methods to analyze the methylome of a specific type of neuron in the mouse brain, and found high-quality data. Additionally, the study identified cell type-specific regulatory regions and compared methylomes of different cell types.
DNA methylation plays essential roles in various cellular processes. Next-generation sequencing has enabled us to study the functional implication of DNA methylation across the whole genome. However, this approach usually requires a substantial amount of genomic DNA, which limits its application to defined cell types within a discrete brain region. Here, we applied two separate protocols, Accel-NGS Methyl-Seq (AM-seq) and Enzymatic Methyl-seq (EM-seq), to profile the methylome of D2 dopamine receptor-expressing medium spiny neurons (D2-MSNs) in mouse nucleus accumbens (NAc). Using 40 ng DNA extracted from FACS-isolated D2-MSNs, we found that both methods yielded comparably high-quality methylome data. Additionally, we identified numerous unmethylated regions (UMRs) as cell type-specific regulatory regions. By comparing the NAc D2-MSN methylome with the published methylomes of mouse prefrontal cortex excitatory neurons and neural progenitor cells (NPCs), we identified numerous differentially methylated CpG and non-CpG regions. Our study not only presents a comparison of these two low-input DNA whole genome methylation profiling protocols, but also provides a resource of DNA methylome of mouse accumbal D2-MSNs, a neuron type that has critical roles in addiction and other neuropsychiatric disorders.
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