The Molecular Mechanisms Underlying Basolateral Amygdala Neuron Development

基底外侧杏仁核神经元发育的分子机制

• 批准号: 10044197
• 负责人: Rothem Kovner
• 金额: $ 6.79万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-12-01 至 2023-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10044197
• 关键词: Adult; Amygdaloid structure; Behavior; Biological Assay; Brain region; Cell Nucleus; Cells; ChIP-seq; Complex; Cues; Data; Data Set; Development; EMSA; Electrophoretic Mobility Shift Assay; Emotional; Emotions; Etiology; Event; Functional disorder; Gene Expression Profile; Genetic; Genetic Models; Genetic Transcription; Genomics; Genotype; Glutamates; Human; In Situ Hybridization; Interneurons; Knockout Mice; Laboratories; Lateral; Learning; Light; Luciferases; Memory; Molecular; Motivation; Mus; Negative Valence; Neurodevelopmental Disorder; Neurons; Nuclear RNA; Output; Participant; Play; Population; Positive Valence; Prefrontal Cortex; Process; Regulator Genes; Rewards; Role; Small Nuclear RNA; Social Behavior; Source; Stimulus; Stream; Structure; Techniques; Testing; Time; Validation; Work; cell type; chromatin immunoprecipitation; conditional knockout; excitatory neuron; experimental study; member; migration; mouse genetics; neuron development; neuropsychiatric disorder; prenatal; programs; response; transcription factor; transcriptome sequencing; transcriptomics

The amygdala is a formation of interconnected nuclei essential for encoding the emotional and motivational components of behavior, learning, and memory. Perturbations of amygdala development are thought to contribute to many neuropsychiatric disorders, but neither the timing of nor the precise cellular substrates subject to these perturbations are well understood. Consequently, assessing the molecular identities and developmental programs governing the amygdala is essential for understanding the etiology of neuropsychiatric disease. Here, I propose to decipher the gene regulatory networks necessary for the development of the basolateral amygdala complex (BLA), a group of nuclei comprised largely of glutamatergic cells that serve as the primary functional input to the amygdala. Studies from our laboratory have identified transcription factors that play critical roles in the development of glutamatergic neurons, similar to those that populate the amygdala. In this proposal I will focus on one transcription factor, characterizing its role in the migration and survival of different types of BLA glutamatergic neurons and identify its downstream effectors. To do so, I will use a conditional knockout mouse and conduct single nuclear RNA sequencing (snRNA-Seq), chromatin immunoprecipitation followed by sequencing (ChIP-seq), and validation experiments of candidate downstream effectors. Cellular, molecular, and genomic techniques are well established in the Sestan laboratory, and this expertise will allow me to both elucidate the molecular processes governing BLA glutamatergic projection neuron specification and identity and aid in deciphering the relevant developmental context underlying numerous neuropsychiatric disorders.

杏仁核是由相互连接的核团组成的，对编码行为、学习和记忆的情绪和动机成分至关重要。杏仁核发育的扰动被认为是许多神经精神障碍的原因，但这些扰动的时间和精确的细胞底物都不是很清楚。因此，评估控制杏仁核的分子特性和发育程序对于了解神经精神疾病的病因学至关重要。在这里，我建议破译杏仁基底外侧复合体(BLA)发育所需的基因调控网络，BLA是一组核团，主要由谷氨酸能细胞组成，充当杏仁核的主要功能输入。我们实验室的研究已经确定了在谷氨酸能神经元的发育过程中发挥关键作用的转录因子，类似于杏仁核中的转录因子。在这个提案中，我将重点关注一个转录因子，描述它在不同类型的BLA谷氨酸能神经元的迁移和存活中的作用，并确定它的下游效应器。为此，我将使用一只有条件的基因敲除小鼠，进行单核RNA测序(SnRNA-Seq)、染色质免疫沉淀后测序(ChIP-SEQ)以及候选下游效应器的验证实验。细胞、分子和基因组技术在Sestan实验室建立得很好，这种专业知识将使我能够阐明支配BLA谷氨酸能投射神经元规格和身份的分子过程，并帮助破译许多神经精神疾病背后的相关发育背景。