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: 8628623
• 负责人: Susan M. Dymecki
• 金额: $ 63.76万
• 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/8628623
• 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; 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; improved; in vivo; innovation; male; motivated behavior; nerve supply; neural circuit; neurochemistry; neuronal cell body; new technology; novel; postsynaptic; public health relevance; receptor; receptor expression; response; social; therapeutic target; tool; transcription factor; transcriptome sequencing

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 regu- lated 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 neu- rons 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 parame- ters change across the life span, perhaps bearing on human age-related propensity for impulsivity, aggres- sion, and substance abuse? In Aim 1, we will identify functional properties of the Drd1a and Drd2 5-HT neu- ron 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 mo- lecular 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 re- gions under neuron-subtype control - the goal of Aim 2. Functional postsynaptic connections will also be ex- plored, 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 si- lence 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 ap- proaches 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.

拟议的研究带来了尖端的遗传工具，以了解理解不足的神经回路 说谎的侵略和动机行为。我们最近的进步表明，这些小鼠的行为是对的 由两个分子不同的小脑血清素能（5-HT）神经元的小节盖结构：一个子集，〜3,000 神经元，通过D1多巴胺（DA）受体的表达在5-HT神经元之间唯一定义（DRD1A） D2 DA受体（DRD2基因），Gene）和另一个神经元。这些结果与 现在可以使用启用遗传工具，例如强大的锋利楔形，并可以“​​打破打开”并访问 电路，细胞性质和分子途径，这些后果行为为基础。我们在这里 提出以四个目的的形式应用此“楔形”，以回答：什么是细胞和分子特性 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，这是这些5-ht neu- RON对DA的反应 - 一种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？做这些选言 - 在整个寿命中的变化，也许与人类年龄相关的冲动性倾向，敏捷性，aggres- sion和药物滥用？在AIM 1中，我们将确定drd1a和drd2 5-ht neu-的功能特性 RON亚型通过转录分析（RNA-SEQ）和电生理记录。这项工作已启用 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类和mo- lecular分析，用于全细胞记录。这些相同的遗传工具不仅可以访问 5-HT神经元亚型，也适用于轴突和末端，从而可以精确地鉴定靶脑的重复 在神经元辅助控制下的gions - 目标2的目标。功能性突触后连接也将被视为 充满了，我们的交叉遗传标记工具将前所未有的分辨率授予经典道 追踪技术以及涉及跨突触示踪剂的尖端病毒方法。因此，目标2 将定义，标记并允许“侵略性抗衡”突触后神经元的分子表征 在这些电路中下游。在AIM 3中，我们将更深入地探索这些行为方面。 两个5-HT神经元的亚型，如果它们的贡献在整个寿命中有所不同。类似的亚型特异性沉默 方法将与基础侵略研究一样采用，但现在其他社会行为和 神经功能将被查询。在AIM 4中，我们将使用药物遗传学（Dreadds）瞬时 在“童年”期间，每个drd 5-ht神经元亚型的透镜，询问是否发生持久的变化 正如人类研究所预测的那样 通过5-HT系统，童年应力和成人病理攻击的遗传易感性。我们的ap- 在技​​术上和概念上都是创新的，并且是发现新的，潜在的基础 行为选择性，适合年龄的治疗学。结果迫使5-HT系统组织的重新定义。