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用于检测神经元抑制的反向活动标记（IAM） （广义定义为神经元活动的减少）。立即早期基因的转录 （IEG）如 c-Fos 和 Arc 已被最广泛地用于将神经元活动转化为稳定的、 可追踪的组织学标签，允许结构和功能询问。现有活动定位 通过直接检测 IEG 或工程 IEG 启动子的方法，针对 检测神经活动的持续增加。然而，它们通常不太有效 标记神经元活动的抑制。因此，为了更好地理解双向大脑 活动，重要的是有一组新的标记来标记神经元活动的减少 与传统的 IEG 相反，我们在这里将其称为反向活动标记。 我们的目标是开发基于蛋白质翻译后修饰 (PTM) 的 IAM，众所周知 快速、双向且可追踪。我们假设如果我们能够逆向识别 PTM 通过无偏筛选与神经元激活相关，这些变化可以发展为 IAM 报告行为动物的神经抑制。我们建立了独创的光遗传学- 蛋白质组学筛选平台，从中我们发现丙酮酸的磷酸化 脱氢酶 E1 亚基 Alpha 1 或 pPDH 与神经元活动呈负相关。我们的中央 假设是为了检验pPDH是否可以作为第一个反映神经活动抑制的IAM 体外和体内。该方法开发的目标是将 IAM 与全脑清算相结合， 光片成像和多重标记，使细胞 ID 兼容工具能够进行无偏分析 全脑抑制。我们组建了一支具有公认专业知识的研究团队 活动依赖性工具开发、电路图谱、电生理学和蛋白质组学以及 行为。这些新颖工具的开发和传播将为我们带来新的视角。 了解大脑的回路动力学。

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。

• 发表时间: 2023-03-14
• 作者: Yang, Dong;Wang, Yu;Qi, Tianbo;Zhang, Xi;Shen, Leyao;Ma, Jingrui;Pang, Zhengyuan;Lal, Neeraj K;McClatchy, Daniel B;Wang, Kristina;Xie, Yi;Polli, Filip;Maximov, Anton;Augustine, Vineet;Cline, Hollis T;Yates, John R;Ye, Li
• 通讯作者: Ye, Li