ATAC-ing dopaminergic cell identity with single-cell resolution

多巴胺神经元是大脑中奖赏通路的关键组成部分。历史上，多巴胺神经元 被认为是一个相对同质的人口介导的强化信号的关联， 食物、水和生殖与感官刺激相一致。然而，最近的研究表明， 神经元形成具有不同生理特征、神经投射和生物功能的亚群。 这些不同的亚群在成瘾周期的进展中也有不同的作用，从使用到使用。 滥用、禁欲和复发。这项应用的目标是设计新的遗传工具， 精确操纵多巴胺回路的不同亚群，特异性低至单对， 神经元为此，我们将首先应用一种新技术，即单细胞ATAC-seq，来确定所有开放的 果蝇中约250个多巴胺神经元中每一个的染色质/可接近的DNA增强子元件 个脑袋第二，我们将确定哪些开放增强子片段或其组合将独特地 识别单个多巴胺细胞/细胞类型。第三，基于这一分析，我们将设计许多新的 基因工具，并测试它们的体内功效和特异性。虽然这些目标是线性的， 相互依存，赠款整体应用单细胞ATAC-seq的新技术，以“打破新的 地面”和“加快发现的步伐，推进成瘾研究”。该提案基本上测试了 这种无偏见的全基因组方法可以用来产生新的工具， 精准干预因为该方法本身可以扩展，应用于任何细胞类型，并转化为 哺乳动物，这项申请完全符合尖端基础研究奖的精神 （CEBRA）机制，将“支持高风险、高影响的研究”（PAR-18-437）。

Dopamine neurons are a critical component of reward pathways in the brain. Historically, dopamine neurons were considered a relatively homogeneous population mediating association of reinforcement signals from food, water, and reproduction with coinciding sensory stimuli. However, recent studies have shown that these neurons form subpopulations with distinct physiological profiles, neural projections, and biological functions. These distinct subpopulations also have different roles in the progression of the addiction cycle, from use to abuse, abstinence and relapse. The goal of this application is to engineer novel genetic tools that will allow precise manipulation of distinct subpopulations of dopamine circuits, with specificity down to single pairs of neurons. To this end we will first apply a new technique, single-cell ATAC-seq, to determine all open chromatin/accessible DNA enhancer elements of every one of the ~250 dopamine neurons in the Drosophila brain. Second, we will determine which open enhancer fragments, or combinations thereof, will uniquely identify single dopamine cells/cell-types. And third, based on this analysis, we will engineer numerous new genetic tools and test them for their in vivo efficacy and specificity. While these Aims are linear and fully interdependent, the grant overall applies the very new technique of single-cell ATAC-seq to “break new ground” and “accelerate the pace of discoveries to advance addiction research”. The proposal essentially tests the hypothesis that this unbiased, genome-wide approach can be harnessed to generate new tools for precision intervention. Because the approach itself can be scaled, applied to any cell type, and translated to mammals, this application is fully consistent with the spirit of the Cutting-Edge Basic Research Awards (CEBRA) mechanism, which will “support high-risk, high impact research” (PAR-18-437).