In vivo nuclear capture and molecular profiling identifies Gmeb1 as a transcriptional regulator essential for dopamine neuron function.
In vivo nuclear capture and molecular profiling identifies Gmeb1 as a transcriptional regulator essential for dopamine neuron function.
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体内核捕获和分子分析将 Gmeb1 确定为多巴胺神经元功能必需的转录调节因子。
DOI：
10.1038/s41467-019-10267-0
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发表时间：
2019
影响因子：
16.6
通讯作者：
Zhang,Yi
中科院分区：
文献类型：
--
作者：
Tuesta,LuisM;Djekidel,MohamedN;Chen,Renchao;Lu,Falong;Wang,Wengang;Sabatini,BernardoL;Zhang,Yi
中脑多巴胺（mDA）神经元在奖赏信号传导中起着核心作用，并广泛涉及精神和神经退行性疾病。为了了解mDA神经元如何执行这些功能，了解mDA特异性基因是如何被调节的是很重要的。然而，哺乳动物大脑的细胞异质性对获得这种理解提出了重大挑战。为此，我们开发了一种基于病毒的方法来标记和捕获mDA细胞核，用于转录组（RNA-Seq）和低输入染色质可及性（liDNase-Seq）分析，然后通过预测建模来确定mDA神经元的推定转录调节因子。使用这种方法，我们确定了gmeb1，这是一种转录因子，预计可以调节thanddat的表达，thanddat分别是多巴胺合成和再摄取的关键基因。mDA神经元中gmeb1的敲低导致thanddat的下调，以及严重的运动缺陷。因此，本研究确定了gmeb1是mDA基因表达和功能的主要调控因子，并为鉴定细胞类型特异性转录调控因子提供了一种通用方法。
Midbrain dopamine (mDA) neurons play a central role in reward signaling and are widely implicated in psychiatric and neurodegenerative disorders. To understand how mDA neurons perform these functions, it is important to understand how mDA-specific genes are regulated. However, cellular heterogeneity in the mammalian brain presents a major challenge to obtaining this understanding. To this end, we developed a virus-based approach to label and capture mDA nuclei for transcriptome (RNA-Seq), and low-input chromatin accessibility (liDNase-Seq) profiling, followed by predictive modeling to identify putative transcriptional regulators of mDA neurons. Using this method, we identifiedGmeb1, a transcription factor predicted to regulate expression ofThandDat, genes critical for dopamine synthesis and reuptake, respectively.Gmeb1knockdown in mDA neurons resulted in downregulation ofThandDat, as well as in severe motor deficits. This study thus identifiesGmeb1as a master regulator of mDA gene expression and function, and provides a general method for identifying cell type-specific transcriptional regulators.