Cell Type Specific Genomic and Functional Dissection of Fear-Off Amygdala Pathways

恐惧杏仁核通路的细胞类型特异性基因组和功能解剖

基本信息

批准号：
10748055
负责人：
VADIM BOLSHAKOV
金额：
$ 3.99万
依托单位：
MCLEAN HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-01-01 至 2026-05-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10748055
关键词：
Address Adolescent Adult Aggressive behavior Amygdaloid structure Behavior Behavioral Bite Brain CRH gene Calcium Cell Nucleus Cell physiology Cells Chronic stress Conditioned Stimulus Cues Development Disease Disinhibition Dissection Extinction Fiber Fluorescence Fright Functional disorder Genetic Genomics Hippocampus Home Image Immunohistochemistry Individual Learning Maintenance Medial Mediating Memory Modeling Molecular Mus National Institute of Mental Health Neural Pathways Neurons Operative Surgical Procedures Pathologic Pathway interactions Patients Pattern Photometry Physiological Population Post-Traumatic Stress Disorders Prefrontal Cortex Preparation Proteins Protocols documentation RNA Reporter Research Domain Criteria Research Priority Rodent Role Saline Social Controls Social Interaction Stimulus Strategic Planning Stress Synapses Territoriality Testing Therapeutic Time Tissues Trauma United States National Institutes of Health Work anxiety-related disorders cell type clinical diagnosis conditioned fear designer receptors exclusively activated by designer drugs effective therapy experience experimental study fear memory fighting learning extinction lens male memory consolidation memory recall memory retention mouse model neural circuit novel paired stimuli pharmacologic rational design receptor recruit response social social defeat social stress tool trauma exposure violence exposure

项目摘要

Fear-related disorders such as Post-Traumatic Stress Disorder (PTSD) are often characterized by an inability to inhibit and extinguish fear memories leading to pathological expression of fear-related behaviors. For progress to occur with targeted rationally-designed therapeutic approaches, a greater understanding of the neural circuitry mediating fear inhibition and extinction is needed. This proposal utilizes cutting-edge, cell- type specific approaches targeting circuits that control amygdala fear inhibition and extinction, via medial prefrontal cortex (mPFC) and ventral hippocampus (vHPC) cell-type specific neural pathways, to align with NIMH research priorities by cutting across RDoC domains in the NIH strategic plan for identifying the pathophysiology of fear-related disorders. It is critical that we understand the role of specific cell types projecting to the amygdala supporting fear inhibition and fear extinction learning. This Competitive Renewal expands our prior work dissecting function of cell-type-specific mechanisms in the amygdala that differentially mediate fear and extinction. In addition to other neuronal subtypes, our prior work dissected roles of the CRF and Thy1-specific neuronal populations within the mouse Basolateral Amygdala (BLA) nuclei, demonstrating distinct molecular and physiological functions underlying fear and extinction pathways. Here we aim to extend this work using a variety of currently available intersectional circuit dissection tools, to understand the role of medial prefrontal cortex (mPFC) and ventral hippocampus (vHPC) projections in regulating amygdala CRF and Thy1 populations. We predict that this approach will identify novel pharmacological targets for fear inhibition and extinction, pursuing new pathways for fear-related anxiety disorders. Our central hypothesis is that the specific pathways within the mPFC and vHPC to BLA circuits, involving projections to fear-controlling amygdala CRF- and Thy1-positive cells, contribute to the mechanisms of fear retention. Targeting these specific pathways will provide greater understanding of fear inhibitory control. This hypothesis will be tested through the following Specific Aims: 1) Explore the role of mPFC and vHPC projections to CRF and Thy1 cells in amygdala in control of fear and extinction. 2) Identify activity patterns in mPFC and vHPC neurons projecting to fear-controlling cells in amygdala using GCaMP miniscope and fiber photometry. 3) Explore synaptic and network-level mechanisms of repeated stress-triggered fear renewal, focusing on mPFC and vHPC projections to amygdala CRF and Thy1 positive neurons, respectively. 4) Perform cell type specific RNA profiling of amygdala-projecting mPFC and vHPC neurons (both CRH/Thy1 targeted cells and engram activity dependent cells) with and without chronic stress. The identification of novel targets will advance our understanding of circuitry underlying fear behaviors and will provide unique avenues for therapeutics.

与恐惧相关的疾病，例如创伤后应激障碍 (PTSD)，其特征通常是无法抑制和消除恐惧记忆，导致恐惧相关行为的病态表达。为了通过合理设计的有针对性的治疗方法取得进展，需要更深入地了解需要介导恐惧抑制和消退的神经回路。该提案利用了尖端的细胞技术针对控制杏仁核恐惧抑制和消退的电路的类型特定方法，通过内侧前额皮质 (mPFC) 和腹侧海马 (vHPC) 细胞类型特定的神经通路，以与 NIMH 的研究重点是通过跨越 NIH 战略计划中的 RDoC 领域来确定恐惧相关疾病的病理生理学。至关重要的是，我们了解投射到杏仁核的特定细胞类型支持恐惧的作用抑制和恐惧消退学习。此次竞争性更新扩展了我们之前的工作剖析功能杏仁核中差异介导恐惧和灭绝的细胞类型特异性机制。此外其他神经元亚型，我们之前的工作剖析了 CRF 和 Thy1 特异性神经元群体的作用在小鼠基底外侧杏仁核（BLA）核内，表现出独特的分子和生理学恐惧和灭绝途径的潜在功能。在这里，我们的目标是使用各种方法来扩展这项工作目前可用的交叉电路解剖工具，以了解内侧前额叶皮层的作用 (mPFC) 和腹侧海马 (vHPC) 预测在调节杏仁核 CRF 和 Thy1 群体中的作用。我们预测这种方法将确定抑制恐惧的新药理学靶点灭绝，寻找与恐惧相关的焦虑症的新途径。我们的中心假设是 mPFC 和 vHPC 内通往 BLA 回路的特定通路，涉及对恐惧控制的预测杏仁核 CRF 和 Thy1 阳性细胞有助于恐惧保留机制。瞄准这些特定的途径将有助于更好地理解恐惧抑制控制。这个假设将是通过以下具体目标进行测试： 1) 探索 mPFC 和 vHPC 预测对 CRF 和杏仁核中的 Thy1 细胞控制着恐惧和灭绝。 2) 识别 mPFC 和 vHPC 的活动模式使用 GCaMP 微型显微镜和光纤光度测定法将神经元投射到杏仁核中的恐惧控制细胞。 3）探索重复压力引发的恐惧更新的突触和网络水平机制，重点关注 mPFC 和 vHPC 分别投射到杏仁核 CRF 和 Thy1 阳性神经元。 4) 进行细胞类型杏仁核投射 mPFC 和 vHPC 神经元（CRH/Thy1 靶向细胞和印迹活动依赖细胞）有或没有慢性压力。新目标的识别将增进我们对恐惧行为背后的电路的理解，并将提供独特的途径疗法。