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: 10542445
• 负责人: Maria Medalla
• 金额: $ 20.63万
• 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 2024-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10542445
• 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; Regulation; Rest; Role; Signal Pathway; Signal Transduction; Slice; Sorting; Stress; Structure; Synapses; Techniques; Therapeutic; Tissues; Tracer; Visualization; With laterality; 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; transcriptomic profiling; 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 前扣带皮层 (ACC) 与外侧前额叶皮层 (LPFC) 和边缘系统紧密相连 杏仁核（AMY）等结构，形成对认知和情感处理重要的通路。 人类神经影像学研究表明 ACC 对 LPFC 和 AMY 表现出差异控制 通过反相关激活模式调节不同的边缘和认知功能静息状态网络。 控制唤醒和压力的上行脑干神经调节途径影响神经化学物质 通过调节内在兴奋性和突触来调节这些皮质边缘网络中的环境和活动 特定区域、层和细胞类型中的信号传导。了解迄今未知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——在灵长类动物 ACC 中，一个关键的皮质边缘中心，将使我们能够识别新的生物标志物 神经化学失衡和细胞损伤导致容易受到情绪压力。最近的 单细胞 RNA 测序 (scRNAseq) 的进步使得能够揭示分子定义的神经元 小鼠皮质中的亚类。然而，我们对特定于层的方式的理解存在很大的知识差距 分子神经元身份影响电生理特性和神经元互连的形成 在灵长类动物和人类中。该提案的总体假设是层板特异性 ACCAMY 和 ACCLPFC 投射神经元具有独特的分子特征，这些分子特征是其神经调节信号传导的基础 兴奋性和突触连接的特性和调节。我们建议确定基因表达 个体层特异性 ACC 细胞类型和投射神经元的概况，重点关注差异表达 与唤醒和压力相关的受体和信号通路。我们将把神经束追踪与批量结合起来 和 scRNAseq 以及体外电生理技术在成年恒河猴 (Macaca mulatta) 中的应用 目前资助的赠款（R01MH116008）。我们将使用最先进的 Patch-Seq 方法进行转录组学研究 对单个电生理学特征层和目标特异性神经元进行分析。差异化 通过单细胞转录组学鉴定的表达基因将使用 RNAscope 进行验证 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。我们将具体阐明 ACCAMY 和 ACCLPFC 的分子和生理特性 与调节认知、唤醒和情绪压力有关的投射神经元。调查结果来自 拟议的研究将构成一个更大的研究计划的基础，以调查这些神经信号传导如何 特性与神经化学失衡和对不利条件（例如氧化应激）的反应有关 和炎症。拟议的研究将揭示层状和功能性的分子基础 调节情感行为的 ACC 回路的多样性，对于理解具有广泛的治疗意义 与压力相关的疾病的易感性。