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来了解神经回路 谎言攻击和动机行为。我们最近的进展表明，小鼠的这些行为是有规律的 由大脑血清素能 (5-HT) 神经元的两个分子上不同的小亚群引起：一个亚群，约 3,000 个 神经元，通过表达 D1 多巴胺 (DA) 受体 (Drd1a) 在 5-HT 神经元中被独特定义 基因），另一个约 1,000 个神经元，由 D2 DA 受体（Drd2 基因）控制。这些结果与 启用遗传工具，如强力锋利的楔子，现在可用于“打开”并访问 这些结果行为背后的电路、细胞特性和分子途径。在这里我们 建议以四个目的的形式应用这个“楔子”来回答：什么是细胞和分子特性 这些行为关键型 5-HT 神经元有何独特之处？正如受体表达所表明的，这些 5-HT 是中性的吗？ rons 对 DA（一种通常与大脑奖励系统相关的神经化学物质）有反应吗？ 这些亚型通过什么前脑回路调节攻击性和动机？执行这些参数- 它们在整个生命周期中都会发生变化，也许与人类与年龄相关的冲动、攻击性倾向有关。 和药物滥用？在目标 1 中，我们将确定 Drd1a 和 Drd2 5-HT neu- 的功能特性 通过转录分析（RNA-seq）和电生理记录来识别 ron 亚型。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，适用于神经元亚型排序和mo- 细胞分析和全细胞记录。这些相同的遗传工具不仅提供了进入细胞体的途径 5-HT 神经元亚型，还包括轴突和末梢，从而可以精确识别目标大脑重新 神经元亚型控制下的神经元 - 目标 2 的目标。功能性突触后连接也将是前- 值得探索的是，我们的交叉基因标记工具为经典领域提供了前所未有的解决方案- 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、标记并允许对“攻击相关”突触后神经元进行分子表征 这些电路的下游。在目标 3 中，我们将更深入地探索这些因素所调节的行为方面 两种 5-HT 神经元亚型以及它们的贡献在整个生命周期中是否有所不同。类似的亚型特异性沉默 将采用与基础攻击研究相同的方法，但现在增加了社会行为和 神经功能将受到询问。在目标 4 中，我们将使用药物遗传学 (DREADD) 来瞬时筛选 研究“童年”期间的每个 Drd 5-HT 神经元亚型，询问是否会发生持久的变化，从而导致 正如人类研究所预测的那样，成年后的过度攻击性和社会动机的改变 通过 5-HT 系统、童年压力和成人病理性攻击行为产生的遗传倾向。我们的应用程序- 这些方法在技术和概念上都具有创新性，是发现新的、有潜力的基础 行为选择性、适合年龄的治疗方法。结果迫使我们重新定义 5-HT 系统组织。