Brain-wide mapping of neuronal inhibition by novel inverse activity markers

通过新型反向活动标记物绘制全脑神经元抑制图谱

• 批准号: 10639977
• 负责人: Li Ye
• 金额: $ 301.35万
• 依托单位: SCRIPPS RESEARCH INSTITUTE, THE
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-05-02 至 2026-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10639977
• 关键词: Agreement; Animals; Antibodies; Behavior; Benchmarking; Biochemistry; Brain; Brain Mapping; Brain region; Cells; Citric Acid Cycle; Collaborations; Communities; Data; Detection; Development; Disease; Electrophysiology (science); Engineering; Enzymes; FOS gene; Fluorescence; Future; General Anesthesia; Genetic Transcription; Goals; Histological Labelings; Hypothalamic structure; Image; Immediate-Early Genes; Immunofluorescence Immunologic; Label; Lateral; Light; Mainstreaming; Maps; Methods; Molecular Target; Neural Inhibition; Neurons; Neurosciences; Oxidoreductase; Phosphorylation; Population; Post-Translational Protein Processing; Proteomics; Pyruvate; Pyruvate Dehydrogenase E1; Reporting; Research Personnel; Side; Specificity; Stains; System; Testing; Translating; Visual; activity marker; cell type; experience; high throughput screening; in vitro activity; in vivo; method development; nervous system disorder; neural; novel; novel marker; optogenetics; phosphoproteomics; promoter; screening; sensor; success; temporal measurement; tool; tool development

Abstract This project aims to develop the first Inverse Activity Marker (IAM) for detecting neuronal inhibition (broadly defined as the decrease of neuronal activities). The transcription of immediate early genes (IEGs) like c-Fos and Arc has been the most widely used for translating neuronal activity into stable, trackable histological labels to allow structural and functional interrogations. Existing activity targeting methods, either through direct detection of IEGs or engineered IEG promoters, are optimized for detecting the sustained increase of neural activity. However, they are generally less effective for labeling the inhibition of neuronal activity. Therefore, to better understand the bi-directional brain activities, it is important to have a set of new markers to label the decrease of neuronal activity opposite to the conventional IEGs, which we propose here as the Inverse Activity Marker. We aim to develop IAMs based on protein post-translational modifications (PTMs), which are known to be rapid, bi-directional, and trackable. We hypothesize that if we can identify PTMs inversely correlated with neuronal activation through unbiased screens, these changes could be developed into IAMs to report neural inhibition in behaving animals. We established an original optogenetic- proteomics screening platform, from which we discovered that the phosphorylation of pyruvate dehydrogenase E1 subunit Alpha 1 or pPDH inversely correlated with neuronal activity. Our central hypothesis is to test whether pPDH can serve as the first IAM to reflect the inhibition of neural activity in vitro and in vivo. The method development goal is to integrate IAMs with whole-brain clearing, lightsheet imaging, and multiplexed labeling to enable a cell-ID compatible tool for unbiased profiling of brain-wide inhibition. We assembled a team of investigators with well-recognized expertise in activity-dependent tool development, circuit mapping, electrophysiology, and proteomics and behaviors. The development and dissemination of these novel tools will bring new perspectives to understanding the circuit dynamics of the brain.

抽象的 该项目旨在开发第一个反活动标记（IAM）来检测神经元抑制 （广义定义为神经元活性的减少）。直接早期基因的转录 （IEG）像C-Fos和ARC一样，是将神经元活性转化为稳定的最广泛使用的 可跟踪的组织学标签，允许结构和功能性询问。现有的活动定位 通过直接检测IEG或工程IEG启动子的方法，方法是优化的 检测神经活动的持续增加。但是，它们通常对 标记神经元活性的抑制作用。因此，更好地了解双向大脑 活动，重要的是要有一组新标记来标记降低神经元活动 与传统的IEG相反，我们在这里建议将其作为反活动标记。 我们旨在基于蛋白质后翻译（PTM）开发IAM，这是已知的 快速，双向且可追踪。我们假设，如果我们可以否认PTM 与神经元激活通过公正的屏幕相关，可以将这些变化发展为 IAMS报告行为动物的神经抑制作用。我们建立了原始的光遗传学 - 蛋白质组学筛选平台，我们从中发现丙酮酸的磷酸化 脱氢酶E1亚基1或PPDH与神经元活性成反比。我们的中心 假设是测试PPDH是否可以作为反映神经活动抑制的第一个IAM 体外和体内。方法开发目标是将IAM与全脑清除融化， 灯显示成像和多重标记，以实现一个兼容的细胞ID工具，以无偏分析 大脑抑制作用。我们组建了一组调查员，拥有公认的专业知识 活动依赖性工具开发，电路映射，电生理学和蛋白质组学以及 行为。这些新工具的发展和传播将为 了解大脑的电路动力学。

项目成果

Phosphorylation of pyruvate dehydrogenase marks the inhibition of in vivo neuronal activity.

丙酮酸脱氢酶的磷酸化标志着体内神经元活动的抑制。

• DOI: 10.1101/2023.03.13.532494
• 发表时间: 2023
• 期刊: bioRxiv : the preprint server for biology
• 作者: Yang,Dong;Wang,Yu;Qi,Tianbo;Zhang,Xi;Shen,Leyao;Ma,Jingrui;Pang,Zhengyuan;Lal,NeerajK;McClatchy,DanielB;Wang,Kristina;Xie,Yi;Polli,Filip;Maximov,Anton;Augustine,Vineet;Cline,HollisT;Yates,JohnR;Ye,Li
• 通讯作者: Ye,Li