FACS-array?based cell purification yields a specific transcriptome of striatal medium spiny neurons in a murine Huntington disease model
FACS-array?based cell purification yields a specific transcriptome of striatal medium spiny neurons in a murine Huntington disease model
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DOI：
10.1074/jbc.ra120.012983
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发表时间：
2020-07-17
影响因子：
4.8
通讯作者：
Nukina, Nobuyuki
中科院分区：
文献类型：
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作者：
Miyazaki, Haruko;Yamanaka, Tomoyuki;Nukina, Nobuyuki
亨廷顿病（Huntington disease，HD）是一种由亨廷顿基因CAG重复序列扩增引起的神经退行性疾病。以往的研究结果表明，转录调控异常的关键机制之一，纹状体中型棘神经元（MSN）变性在HD。然而，由于非MSN细胞的污染，HD病因学或病理学中涉及的一些关键基因可能在常见的表达谱测定中被掩盖。为了深入了解MSN特异性基因表达的变化，在presymptosis R6/2小鼠，一种常见的HD小鼠模型，在这里，我们使用转基因荧光蛋白标记的MSN的纯化，通过流式细胞仪分析基因表达与基因微阵列和比较的结果，这？流式细胞仪与用均质化纹状体样本（STR阵列）获得的结果相同。我们确定了数百个差异表达基因（DEG）和MSN特异性DEG的检测，通过比较结果的FACS阵列与STR阵列。获得的基因组包括在MSN和非MSN脑细胞中普遍表达的基因，并与转录调控和DNA损伤反应相关。我们提出，比较基因表达的方法，使用流式细胞仪阵列可能是有用的揭示基因级联影响的MSN在HD发病机制。
Huntington disease (HD) is a neurodegenerative disorder caused by expanded CAG repeats in theHuntingtingene. Results from previous studies have suggested that transcriptional dysregulation is one of the key mechanisms underlying striatal medium spiny neuron (MSN) degeneration in HD. However, some of the critical genes involved in HD etiology or pathology could be masked in a common expression profiling assay because of contamination with non-MSN cells. To gain insight into the MSN-specific gene expression changes in presymptomatic R6/2 mice, a common HD mouse model, here we used a transgenic fluorescent protein marker of MSNs for purification via FACS before profiling gene expression with gene microarrays and compared the results of this ?FACS-array? with those obtained with homogenized striatal samples (STR-array). We identified hundreds of differentially expressed genes (DEGs) and enhanced detection of MSN-specific DEGs by comparing the results of the FACS-array with those of the STR-array. The gene sets obtained included genes ubiquitously expressed in both MSNs and non-MSN cells of the brain and associated with transcriptional regulation and DNA damage responses. We proposed that the comparative gene expression approach using the FACS-array may be useful for uncovering the gene cascades affected in MSNs during HD pathogenesis.