Gene Expression Programs in Maturing Dopamine Neurons: Temporal Regulation, Subcellular Localization, and Alteration by Amphetamine

成熟多巴胺神经元中的基因表达程序：时间调节、亚细胞定位和安非他明的改变

• 批准号: 10310429
• 负责人: Benjamin Hobson
• 金额: $ 5.18万
• 依托单位: COLUMBIA UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-01-15 至 2023-07-14
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10310429
• 关键词: Academic Medical Centers; Address; Adolescence; Adolescent; Adolescent Development; Adult; Affect; Afferent Neurons; Affinity Chromatography; Algorithms; Amphetamines; Award; Axon; Behavioral; Biochemical; Biology; Brain; Candidate Disease Gene; Cell Death; Cells; Cellular biology; Clinical; Clinical Skills; Cognitive deficits; Complex; Corpus striatum structure; DCC gene; Data; Development; Developmental Gene; Developmental Gene Expression Regulation; Developmental Process; Disease; Dopamine; Dorsal; Drug Addiction; Drug Exposure; Drug usage; Drug user; Embryo; Exposure to; Fellowship; Forebrain Development; Functional disorder; Gene Expression; Gene Expression Profile; Genetic Transcription; Genetic Translation; Goals; Growth Cones; Health; Knowledge; Link; Logic; Mediating; Mentorship; Messenger RNA; Methodology; Midbrain structure; Mus; NTN1 gene; Neurons; Neurosciences; Nucleus Accumbens; Pattern; Pharmacology; Phosphorus 32; Physicians; Physiology; Positioning Attribute; Prefrontal Cortex; Process; Prosencephalon; Protein Biosynthesis; Proteins; Regulation; Regulator Genes; Research; RiboTag; Ribosomes; Risk; Role; Sampling; Scientist; Shapes; Signal Transduction; Slice; Social outcome; Structure; Substance Use Disorder; Systems Biology; Technical Expertise; Testing; Training; Transgenic Mice; Translating; Translational Regulation; Translations; addiction; adolescent drug use; adverse outcome; axon growth; axon guidance; career; cell type; differential expression; dopaminergic neuron; drug of abuse; early adolescence; early onset drug use; experience; extracellular; genome-wide; genome-wide analysis; in vivo; interest; knock-down; mRNA sequencing; nerve supply; netrin receptor; neuron development; neuron loss; neuronal cell body; neuropsychiatric disorder; neuropsychiatry; neurotransmission; new technology; next generation sequencing; novel; novel sequencing technology; postnatal; postnatal development; pre-doctoral; presynaptic; prevent; programs; psychostimulant; ranpirnase; response; retinal neuron; ribosome profiling; skills; small hairpin RNA; synaptogenesis; translatome

Early initiation of drug use is associated with long-term poor social outcomes and higher rates of substance use disorders in adulthood. Altered dopamine (DA) neurotransmission in forebrain circuits is a hallmark of drug addiction and DA inputs to these circuits continue to mature into adolescence. Recent evidence suggests that adolescent drug exposure alters DA axonal maturation and disrupts proper forebrain development, resulting in persistent behavioral alterations that may increase the risk of drug addiction. In order to prevent long-term adverse outcomes related to adolescent drug use, we must understand the mechanisms by which drugs of abuse alter normal development of DA neurons. Postnatal development of DA neurons is complex: a small number of DA neurons must survive two rounds of cell death and mature their axonal innervation in expansive forebrain structures, suggesting tight regulation of developmental gene expression programs. These DA neuron developmental programs may be particularly susceptible to amphetamine, a psychostimulant drug of abuse that causes lasting changes in forebrain DA innervation when administered during early adolescence. This proposal will develop a precise understanding of gene expression in DA neurons across postnatal development and will analyze how early adolescent amphetamine exposure alters normal gene expression patterns. A combination of neuroanatomical, biochemical, next-generation sequencing, and systems biology approaches will be applied to a transgenic mouse that enables DA neuron-specific, genome-wide analysis of translating mRNAs. Aim 1 will identify changes in gene expression associated with 1) key transitional timepoints related to axonal growth and cell death in early postnatal development and 2) response to adolescent amphetamine exposure and prolonged axonal maturation during later postnatal development. Cell type-specific expression data will be used to elucidate and validate master regulator genes controlling DA neuron maturation. Aim 2 will explore the role of subcellular mRNA localization and protein synthesis in maturing DA neurons across the same conditions studied in Aim 1 (i.e., across postnatal development and following amphetamine exposure). The proposed research will enhance our understanding of DA neuronal cell biology and will have far reaching implications for the neurodevelopmental origins of drug addiction. This pre- doctoral fellowship will support rigorous scientific training of the applicant at Columbia University Medical Center. An interdisciplinary mentorship team with unique expertise will provide training in both neuroscience and systems biology, enhancing the applicant’s technical skills and professional development. The applicant will also undergo extensive clinical training, refining general clinical skills and providing field-specific exposure relevant to the applicant’s interest in neuropsychiatry. The activities proposed under this award will provide the applicant with the skills required to develop a successful career as an independent physician-scientist.

早期开始使用药物与长期不良的社会后果和较高的物质使用率有关 成年后的使用障碍。前脑回路中多巴胺（DA）神经传递的改变是药物作用的标志。 成瘾和DA输入这些电路继续成熟到青春期。最近的证据表明 青少年药物暴露改变了DA轴突成熟，破坏了正常的前脑发育，导致 持续的行为改变可能会增加药物成瘾的风险。为了防止长期 与青少年药物使用有关的不良后果，我们必须了解 滥用会改变DA神经元的正常发育。DA神经元的出生后发育是复杂的： 许多DA神经元必须在两轮细胞死亡后存活下来，并在扩张的轴突神经支配中成熟。 前脑结构，表明发育基因表达程序的严格调节。这些DA 神经元发育程序可能特别容易受到安非他明的影响，安非他明是一种精神兴奋剂， 在青春期早期滥用药物，导致前脑DA神经支配的持久变化。 这一建议将发展一个精确的了解基因表达的DA神经元在出生后 并将分析早期青少年安非他明暴露如何改变正常的基因表达 模式.神经解剖学、生物化学、下一代测序和系统生物学的结合 方法将应用于转基因小鼠，使DA神经元特异性，全基因组分析， 翻译mRNA。目的1将确定与以下相关的基因表达的变化：1）关键的过渡 与出生后早期发育中轴突生长和细胞死亡相关的时间点，以及2）对 青少年安非他明暴露和后期出生后发育期间轴突成熟延长。细胞 型特异性表达数据将用于阐明和验证控制DA的主调节基因 神经元成熟目的2：探讨亚细胞mRNA定位和蛋白质合成在细胞凋亡中的作用。 在Aim 1中研究的相同条件下使DA神经元成熟（即，在产后发育中， 安非他明暴露后）。该研究将加深我们对DA能神经细胞的认识 这将对药物成瘾的神经发育起源产生深远的影响。这个预- 博士奖学金将支持申请人在哥伦比亚大学医学院接受严格的科学培训 中心一个具有独特专业知识的跨学科导师团队将提供神经科学 和系统生物学，提高申请人的技术技能和专业发展。申请人 还将接受广泛的临床培训，提高一般临床技能，并提供特定领域的接触 与申请人对神经精神病学的兴趣有关。该奖项下的活动将提供 申请人具有作为独立的物理学家-科学家发展成功职业所需的技能。