Transcriptomic, physiological, and neurochemical profiling of cortico-limbic projection neurons in monkey anterior cingulate cortex

猴子前扣带皮层皮质边缘投射神经元的转录组学、生理学和神经化学分析

• 批准号: 10371649
• 负责人: Maria Medalla
• 金额: $ 24.75万
• 依托单位: BOSTON UNIVERSITY MEDICAL CAMPUS
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-01-01 至 2023-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10371649
• 关键词: Acute; Adult; Affect; Affective; Amygdaloid structure; Anterior; Area; Arousal; Behavior; Biophysics; Brain; Brain Stem; Cells; Cognition; Cognitive; Communication; Data; Dementia; Electrophysiology (science); Emotional; Emotional Stress; Emotions; Environment; Exhibits; Funding; Gene Expression; Gene Expression Profile; Gene Proteins; Genes; Grant; Harvest; Human; Immunohistochemistry; In Situ; In Vitro; Individual; Inflammation; Injections; Ion Channel; Knowledge; Label; Lateral; Link; Macaca mulatta; Mediating; Methods; Molecular; Molecular Profiling; Monkeys; Mood Disorders; Motor; Mus; Nature; Neurons; Oxidative Stress; Pathway interactions; Pattern; Physiological; Population Projection; Predisposition; Prefrontal Cortex; Primates; Property; Receptor Signaling; Regulation; Rest; Role; Signal Pathway; Signal Transduction; Slice; Sorting - Cell Movement; Stress; Structure; Synapses; Techniques; Therapeutic; Tissues; Tracer; age related; cell injury; cell type; cingulate cortex; cortico-limbic circuits; differential expression; experimental study; gene network; hippocampal pyramidal neuron; neural tract; neurochemistry; neuroimaging; neuronal excitability; neuroregulation; neurotransmission; novel marker; patch clamp; patch sequencing; programs; protein expression; receptor; receptor expression; response; single-cell RNA sequencing; stress related disorder; synaptic function; transcriptome sequencing; transcriptomics; Acute; Adult; Affect; Affective; Amygdaloid structure; Anterior; Area; Arousal; Behavior; Biophysics; Brain; Brain Stem; Cells; Cognition; Cognitive; Communication; Data; Dementia; Electrophysiology (science); Emotional; Emotional Stress; Emotions; Environment; Exhibits; Funding; Gene Expression; Gene Expression Profile; Gene Proteins; Genes; Grant; Harvest; Human; Immunohistochemistry; In Situ; In Vitro; Individual; Inflammation; Injections; Ion Channel; Knowledge; Label; Lateral; Link; Macaca mulatta; Mediating; Methods; Molecular; Molecular Profiling; Monkeys; Mood Disorders; Motor; Mus; Nature; Neurons; Oxidative Stress; Pathway interactions; Pattern; Physiological; Population Projection; Predisposition; Prefrontal Cortex; Primates; Property; Receptor Signaling; Regulation; Rest; Role; Signal Pathway; Signal Transduction; Slice; Sorting - Cell Movement; Stress; Structure; Synapses; Techniques; Therapeutic; Tissues; Tracer; age related; cell injury; cell type; cingulate cortex; cortico-limbic circuits; differential expression; experimental study; gene network; hippocampal pyramidal neuron; neural tract; neurochemistry; neuroimaging; neuronal excitability; neuroregulation; neurotransmission; novel marker; patch clamp; patch sequencing; programs; protein expression; receptor; receptor expression; response; single-cell RNA sequencing; stress related disorder; synaptic function; transcriptome sequencing; transcriptomics

ABSTRACT The anterior cingulate cortex (ACC) is heavily interconnected with the lateral prefrontal cortex (LPFC) and limbic structures such as the amygdala (AMY), forming pathways important for cognitive and affective processing. Human neuroimaging studies have shown that the ACC exhibits differential control of LPFC and AMY to modulate distinct limbic and cognitive functional resting state networks with anti-correlated activation patterns. Ascending brainstem neuromodulatory pathways that control arousal and stress influence the neurochemical environment and activity in these cortico-limbic networks through modulating intrinsic excitability and synaptic signaling in specific areas, layers, and cell types. Understanding the as yet unknown cell type-specific molecular signatures of neuromodulatory signaling pathways -- the receptor expression profiles and downstream gene network targets– in the primate ACC, a key cortico-limbic hub, will allow us to identify novel biomarkers for neurochemical imbalance and cellular damage that confer vulnerability to emotional stress. The recent advancement in single-cell RNA sequencing (scRNAseq) has allowed to uncover molecularly-defined neuronal subclasses in mouse cortex. However, there is a big knowledge gap in our understanding of how layer-specific molecular neuronal identity affects electrophysiological properties and the formation of neuronal interconnections in primates and humans. The overall hypothesis of this proposal is that the lamina-specific ACCAMY and ACCLPFC projection neurons have distinct molecular signatures that underlie their neuromodulatory signaling properties and regulation of excitability and synaptic connections. We propose to determine the gene expression profile of individual lamina-specific ACC cell types and projection neurons, focusing on the differential expression of receptors and signaling pathways related to arousal and stress. We will combine neural tract-tracing with bulk and scRNAseq and in vitro electrophysiological techniques in adult rhesus monkeys (Macaca mulatta) from our currently funded grant (R01MH116008). We will use the cutting-edge Patch-Seq method for transcriptomic profiling of individual electrophysiologically-characterized layer- and target- specific neurons. Differentially expressed genes identified via single cell transcriptomics, will be validated using RNAscope and immunohistochemistry (IHC) analysis, and will be correlated with specific functional properties of distinct ACC cell types. We will specifically illuminate the molecular and physiological identity of ACCAMY and ACCLPFC projection neurons that are implicated in regulating cognition, arousal and emotional stress. Findings from the proposed study will form the basis of a larger program of studies to investigate how these neural signaling properties are linked to neurochemical imbalance and responses to adverse conditions, such as oxidative stress and inflammation. The proposed study will unravel the molecular underpinnings of laminar and functional diversity of ACC circuits mediating affective behavior, which has broad therapeutic implications for understanding susceptibility in stress-related disorders.

抽象的 前扣带回皮层（ACC）与外侧前额叶皮层（LPFC）和缘 杏仁核（Amy）等结构，形成对认知和情感处理重要的途径。 人类神经影像学研究表明，ACC对LPFC和Amy表现出差异控制 调节具有反相关激活模式的独特的边缘和认知功能静止状态网络。 控制唤醒和压力影响神经化学的上升脑干神经调节途径 通过调节内在的激动人心和突触 在特定区域，层和细胞类型中发出信号。了解尚未知道的细胞类型特异性分子 神经调节信号通路的特征 - 受体表达谱和下游基因 网络目标 - 在主要ACC中，一个关键的Cortico-limbic Hub，将使我们能够识别出新颖的生物标志物 神经化学失衡和细胞损害，会议易受情绪压力的脆弱性。最近 单细胞RNA测序（SCRNASEQ）的进步已允许发现分子定义的神经元 小鼠皮质中的子类。但是，我们对特定层的理解有很大的知识差距 分子神经元身份影响电生理特性和形成神经元互连 在小学和人类中。该提议的总体假设是椎板特异性的ACPAMY和 ACCLPFC投影神经元具有不同的分子特征，其神经调节信号传导是基于其神经调节信号的 特性和兴奋和突触连接的调节。我们建议确定基因表达 单个椎板特异性ACC细胞类型和投影神经元的曲线，重点是差异表达 接收器以及与唤醒和压力有关的信号通路。我们将结合神经道追踪与批量 我们 目前资助的赠款（R01MH116008）。我们将使用尖端的补丁序列方法进行转录组 分析单个电生理学特异性神经元和靶神经元的分析。差异 通过单细胞转录组学鉴定的表达基因将使用rnascope和 免疫组织化学（IHC）分析，并将与不同ACC的特定功能特性相关 细胞类型。我们将专门阐明ACCAMY和ACP的分子和物理身份 在调节认知，唤醒和情绪压力中暗示的投射神经元。来自 拟议的研究将构成更大的研究计划的基础，以研究这些神经信号如何 特性与神经化学不平衡和对不利条件的反应有关，例如氧化应激 和炎症。拟议的研究将阐明层流和功能的分子基础 ACC电路的多样性介导情感行为，这对理解具有广泛的治疗意义 与压力有关的疾病的敏感性。