Developmental gene networks of 5HT neurons in addiction, aggression, and anxiety

成瘾、攻击性和焦虑中 5HT 神经元的发育基因网络

• 批准号: 8836993
• 负责人: Susan M. Dymecki
• 金额: $ 59.27万
• 依托单位: HARVARD MEDICAL SCHOOL
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-04-15 至 2019-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8836993
• 关键词: Action Potentials; Adolescence; Adult; Adverse effects; Affect; Affective; Age; Aggressive behavior; Alleles; Anxiety; Axon; Behavior; Behavioral; Biological Assay; Brain; Brain region; Cells; Childhood; Cognitive; Development; Developmental Gene; Disease; Dopamine; Dopamine D1 Receptor; Dopamine D2 Receptor; Electrophysiology (science); Emotional; Functional disorder; Gene Expression Profile; Genes; Genetic; Genetic Predisposition to Disease; Goals; Grant; Health; Human; Impulsivity; Individual; Ion Channel; Label; Link; Longevity; Maps; Methods; Molecular; Molecular Profiling; Mood Disorders; Motivation; Mus; Nervous System Physiology; Neurobiology; Neurons; Pathway interactions; Pattern; Pharmacogenetics; Phenotype; Presynaptic Terminals; Property; Prosencephalon; RNA; Research; Resolution; Rewards; Sensorimotor functions; Serotonergic System; Serotonin; Shapes; Slice; Social Behavior; Social Interaction; Sorting - Cell Movement; Stress; Substance abuse problem; Synapses; System; Techniques; Testing; Therapeutic; Tracer; Transgenes; Ursidae Family; Viral; Whole-Cell Recordings; Work; addiction; age related; base; behavior test; brain cell; cell type; critical period; differential expression; emotional behavior; improved; in vivo; innovation; male; motivated behavior; nerve supply; neural circuit; neurochemistry; neuronal cell body; new technology; novel; postsynaptic; postsynaptic neurons; receptor; receptor expression; response; social; therapeutic target; tool; transcription factor; transcriptome sequencing

DESCRIPTION (provided by applicant): The proposed research brings cutting-edge genetic tools to bear on understanding the neural circuitry under- lying aggression and motivated behavior. Our recent progress reveals that these behaviors in mice are regulated by two molecularly distinct, small subsets of brain serotonergic (5-HT) neurons: one subset, ~3,000 neurons, uniquely defined among 5-HT neurons by expression of the D1 dopamine (DA) receptor (Drd1a gene), and the other, ~1,000 neurons, by the D2 DA receptor (Drd2 gene). These results combined with the enabling genetic tools, like a powerfully sharpened wedge, can now be used to "break open" and access the circuitry, cellular properties, and molecular pathways underlying these consequential behaviors. Here we propose applying this "wedge" in the form of four aims to answer: What cellular and molecular properties are unique to these behavior-critical 5-HT neurons? As suggested by receptor expression, are these 5-HT neurons responsive to DA - a neurochemical commonly associated with the reward system of the brain? Through what forebrain circuitry do these subtypes modulate aggression and motivation? Do these parameters change across the life span, perhaps bearing on human age-related propensity for impulsivity, aggression, and substance abuse? In Aim 1, we will identify functional properties of the Drd1a and Drd2 5-HT neuron subtypes by transcriptional profiling (RNA-seq) and electrophysiological recording. This work is enabled through novel genetic tools for neuron subtype-specific marking, suitable for neuron subtype sorting and molecular profiling, and for whole-cell recording. These same genetic tools not only offer access to the soma of a 5-HT neuron subtype, but also to axons and terminals, thus allowing precise identification of target brain regions under neuron-subtype control - the goal of Aim 2. Functional postsynaptic connections will also be explored, with our intersectional genetic marking tools conferring unprecedented resolution to classic tract- tracing techniques as well as to cutting-edge viral approaches that involve trans-synaptic tracers. Thus, Aim 2 will define, label, and allow for molecular characterization of "aggression-relevant" postsynaptic neurons downstream in these circuits. In Aim 3, we will explore more deeply the behavioral facets modulated by these two 5-HT neuron subtypes and if their contributions vary across life span. Similar subtype-specific silencing methods will be employed as in the foundational aggression studies, but now additional social behaviors and neurological functions will be queried. In Aim 4, we will use pharmacogenetics (DREADDs) to transiently silence each Drd 5-HT neuron subtype during "childhood," asking if lasting changes occur that predispose to hyperaggression and altered social motivation in adulthood, as predicted by human studies that associate genetic predisposition via the 5-HT system, childhood stress, and adult pathological aggression. Our approaches are technically and conceptually innovative, and are foundational for discovering new, potentially behavior-selective, age-suitable therapeutics. Results compel a redefinition of 5-HT system organization.

描述（由申请人提供）：拟议的研究带来了尖端的遗传工具，以了解理解神经电路的侵略和动机行为。 Our recent progress reveals that these behaviors in mice are regulated by two molecularly distinct, small subsets of brain serotonergic (5-HT) neurons: one subset, ~3,000 neurons, uniquely defined among 5-HT neurons by expression of the D1 dopamine (DA) receptor (Drd1a gene), and the other, ~1,000 neurons, by the D2 DA receptor (Drd2 gene).这些结果结合了启用遗传工具，例如强大的锐化楔形物，现在可以用来“打破打开”并访问这些结果行为的基础上的电路，蜂窝特性和分子途径。在这里，我们建议以四个目的的形式应用这种“楔形”：哪些细胞和分子特性是这些关键行为5-HT神经元独有的？正如受体表达所暗示的那样，这些5 -HT神经元是否反应DA，一种通常与大脑奖励系统相关的神经化学？这些子类型通过什么前脑电路调节攻击和动机？这些参数在整个生命周期中是否会发生变化，也许与人类年龄相关的冲动，侵略和滥用药物有关？在AIM 1中，我们将通过转录分析（RNA-SEQ）和电生理记录来鉴定DRD1A和DRD2 5-HT神经元亚型的功能性能。这项工作是通过用于神经元亚型特异性标记的新型遗传工具，适用于神经元亚型分选和分子分析以及用于全细胞记录的。这些相同的遗传工具不仅可以访问5-HT神经元亚型的SOMA，而且还可以访问轴突和终端，从而可以精确地识别在神经元补充控制下的目标大脑区域 - AIM 2的目标 - 功能性突触后连接也将通过我们的交易所的遗传分配工具来探索，并构成固定的工具，并定期进行连接的工具，并构成了连接的效果。涉及跨突触示踪剂的尖端病毒方法。因此，AIM 2将定义，标记并允许在这些电路中向下游的“侵略性”的突触后神经元进行分子表征。在AIM 3中，我们将更深入地探索这两个5-HT神经元亚型调节的行为方面，以及它们的贡献在整个寿命中是否有所不同。在基础攻击研究中，将采用类似的亚型特异性沉默方法，但是现在将查询其他社会行为和神经功能。在AIM 4中，我们将使用药物遗传学（Dreadds）在“童年时期”中瞬时沉默每个DRD 5-HT神经元亚型，询问是否发生了持久的变化，是否发生了易感性，并在成年中的社会动机变化，这是通过通过5-HT System System System System Systems Systems Systems Systems，Chilldy Systance and Child and Chirent and Child andication and Child and toction和成人病态侵略的人类研究所预测的。我们的方法在技术上和概念上都是创新的，并且是发现新的，潜在的行为选择性，适合年龄的治疗方法的基础。结果迫使5-HT系统组织的重新定义。