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

• 批准号: 9920213
• 负责人: VADIM BOLSHAKOV
• 金额: $ 66.37万
• 依托单位: MCLEAN HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-07-01 至 2021-07-08
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9920213
• 关键词: Affect; Afferent Neurons; Amygdaloid structure; Behavior; Behavior Therapy; Behavioral; Behavioral inhibition; Biological Assay; Cell Nucleus; Cells; Cre-LoxP; Data; Disease; Dissection; Electrophysiology (science); Event; Exposure to; Extinction (Psychology); FOXP2 gene; Fluorescence-Activated Cell Sorting; Fright; Functional disorder; Genomics; Intercalated Cell; Intervention; Lateral; Measures; Medial; Mediating; Memory; Messenger RNA; MicroRNAs; Molecular; Mus; National Institute of Mental Health; Nature; Neuronal Plasticity; Neurons; Opsin; Outcome; Output; Pathologic; Pathway interactions; Pharmacology; Physiological; Population; Porifera; Post-Traumatic Stress Disorders; Prefrontal Cortex; Process; Regulation; Research Domain Criteria; Research Priority; Rhodopsin; Role; Signal Transduction; Stimulus; Strategic Planning; Synapses; Synaptic Transmission; Synaptic plasticity; Target Populations; Testing; Therapeutic; Training; Trauma; United States National Institutes of Health; Viral; Viral Vector; anxiety-related disorders; cell type; combinatorial; conditioned fear; design; designer receptors exclusively activated by designer drugs; experimental study; fear memory; functional outcomes; in vivo; inducible gene expression; knock-down; learning extinction; neural circuit; notch protein; novel; optogenetics; transcriptome sequencing

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 the amygdala to align with NIMH research priorities by cutting across many of the RDoC 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 within the amygdala supporting fear inhibition and fear extinction learning. It is known that the Basolateral Amygdala (BLA) modulates fear expression via projections to the medial (CeM) and lateral division (CeL) of the central amygdala, in part through modulation via the Intercalated Cell Nuclei of the Amygdala (ITC), which receive inputs from medial prefrontal cortex (mPFC) and act as an inhibitory gate to the CeL. We will target the Thy1, FoxP2, and PKCsubpopulations within the BLA, ITC, CeL, respectively, that are proposed to be associated with fear inhibition and extinction within the amygdala – here termed the ‘Fear-Off’ populations. Through a variety of cell-type specific approaches, we will determine the events underlying the inhibition of fear within three ‘Fear-Off’ cell types in the amygdala. Experiments will functionally and molecularly identify populations of neurons within the BLA, ITC, and CeL that participate in these processes. Our central hypothesis is that the Fear-Off pathway within the BLA-ITC-CeL circuit acts as an mPFC-regulated inhibitory gate facilitating inhibition of fear during extinction training and retention, in part through miRNA regulation of plasticity. Targeting multiple aspects of the Fear-Off pathway will expand our understanding of fear inhibition. These hypotheses will be tested through the following Specific Aims: Aim 1) mPFC Regulation of Amygdala Fear-Off Cells: To determine the differential afferent regulation from mPFC projections to BLA, ITC, and CeA ‘Fear-Off’ neurons through direct manipulation of afferent neurons. Aim 2) Intra-Amygdala Regulation of Fear-Off Cells: To determine the precise role of BLA, ITC, and CeA Fear-Off neurons in regulating fear output circuitry of the amygdala; and Aim 3) miRNA Regulation of Fear-Off Cells: To determine the role of miRNAs on behavior, neuronal function and plasticity within BLA, ITC, and CeA Fear-Off neurons through direct manipulation of miRNA function. We use a combinatorial approach to functionally and molecularly characterize specific target populations of amygdala neurons, which will elucidate important microcircuitry within the amygdala governing fear extinction. The identification of novel targets will advance our understanding of circuitry underlying fear behaviors and will provide unique avenues for therapeutics.

诸如创伤后应激障碍（PTSD）等与恐惧有关的疾病通常以 无法抑制和消除恐惧记忆，从而导致与恐惧相关行为的病理表达。 为了通过有针对性的合理设计的治疗方法进行进展，对 需要神经回路介导恐惧抑制和扩展。该提议利用尖端的细胞 针对针对杏仁核以与NIMH研究优先级保持一致的特定特定方法，通过切割许多 NIH战略计划中的RDOC识别与恐惧有关的疾病的病理生理。 至关重要的是，我们了解杏仁核中特定细胞类型的作用，支持恐惧抑制 并害怕扩展学习。众所周知，基底外侧杏仁核（BLA）通过 中央杏仁核的内侧（CEM）和外侧分裂（CEL）的项目，部分通过调节 通过杏仁核（ITC）的插入细胞核，该细胞核从培养基前额叶皮层接收输入 （MPFC）并充当通往CEL的抑制门。我们将针对Thy1，FoxP2和PKC 在BLA中，分别提议与恐惧抑制和扩展有关 在杏仁核中 - 这里称为“恐惧”人群。 通过多种细胞类型的特定方法，我们将确定抑制的事件 在杏仁核中的三种“恐惧”细胞类型中担心。实验将在功能和分子上识别 参与这些过程的BLA，ITC和CEL中的神经元种群。我们的中心 假设是BLA-ITC-CEL电路中的恐惧途径充当MPFC调节 抑制门在扩展训练和保留期间促进恐惧的抑制，部分通过 miRNA可塑性调节。针对恐惧途径的多个方面将扩大我们的 了解恐惧抑制。 这些假设将通过以下特定目的进行检验：目标1）MPFC调节 杏仁核恐惧关闭细胞：确定从MPFC项目到BLA的差异传入调节， 通过直接操纵传入神经元的直接操纵，ITC和CEA“恐惧”神经元。目标2）杏仁核内 调节恐惧的细胞：确定BLA，ITC和CEA恐惧神经元的精确作用 调节杏仁核的恐惧输出电路；目标3）恐惧细胞的miRNA调节：确定 miRNA在BLA，ITC和CEA恐惧神经元内的行为，神经元功能和可塑性中的作用 通过直接操纵miRNA功能。我们使用组合方法在功能和 分子表征杏仁核神经元的特定目标种群，这将阐明重要 杏仁核内部的微电路，用于恐惧延伸。新目标的识别将推进 我们对恐惧行为的基本电路的理解，并将为治疗提供独特的途径。