The Role of Astrocytes in Cortical Interneuron Development

星形胶质细胞在皮质中间神经元发育中的作用

• 批准号: 8209297
• 负责人: Karen Muller Smith
• 金额: $ 3.86万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-01-06 至 2012-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8209297
• 关键词: Address; Affinity Chromatography; Area; Astrocytes; Attention deficit hyperactivity disorder; Autistic Disorder; Behavior; Behavior Control; Behavioral; Behavioral inhibition; Binding; Bipolar Depression; Brain; Candidate Disease Gene; Cell Maturation; Cell Transplantation; Cells; Cerebral cortex; Child; Childhood; Coculture Techniques; Cognition; Control Animal; Core Facility; Cultured Cells; Data Analyses; Development; Development Plans; Developmental Biology; Disease; Doctor of Philosophy; Dorsal; Educational workshop; Electroporation; Embryo; Environment; Event; Exhibits; FGFR1 gene; Fibroblast Growth Factor; Fibroblast Growth Factor Receptor 1; Fire - disasters; Functional disorder; Future; Gene Expression; Genes; Genetic; Gilles de la Tourette syndrome; Goals; Hand; Health; Hyperactive behavior; In Vitro; Interneurons; Journals; Learning; Light; Medial; Mediating; Mental Depression; Mental disorders; Mentors; Mentorship; Messenger RNA; Methodology; Methods; Microarray Analysis; Molecular; Molecular Genetics; Mus; Mutant Strains Mice; Mutation; Neurobiology; Neurodevelopmental Disorder; Neuroglia; Neurons; Neurosciences; Output; Parvalbumins; Pathway interactions; Pattern; Performance; Perinatal; Phenotype; Pilot Projects; Property; Proteins; Psychiatry; RNA Interference; Research; Research Design; Research Proposals; Research Training; Resources; Reverse Transcriptase Polymerase Chain Reaction; Ribosomes; Role; Schizophrenia; Series; Signal Transduction; Somatostatin; Synapses; System; Tamoxifen; Techniques; Telencephalon; Testing; Time; Training; Training Programs; Transcript; Transgenes; Transgenic Mice; Transgenic Organisms; Translating; Transplantation; Validation; Western Blotting; Work; behavior test; brain cell; calretinin; career; career development; cell cortex; cell type; critical period; developmental neurobiology; excitatory neuron; frontal lobe; in vivo; inhibitory neuron; innovation; insight; knowledge base; mouse model; mutant; neuronal circuitry; neuropsychiatry; neurotrophic factor; novel; post-doctoral training; postnatal; prevent; progenitor; relating to nervous system; research and development; research study; statistics; symposium; tissue culture; vector

DESCRIPTION (provided by applicant): The loss or dysfunction of GABAergic inhibitory interneurons, particularly parvalbumin (PV) expressing interneurons, is implicated in severe psychiatric disorders including schizophrenia, bipolar depression, and Tourette syndrome. PV+ interneurons regulate excitatory neuron output and are important for coordinating brain rhythms so that excitatory neurons can fire in synchrony- properties deemed to be critical for their function in cognition and behavioral control. This proposal describes the career development plan and research aims that Karen M|ller Smith, Ph.D. will achieve during her mentored career development training. The immediate goal of this proposal is to prepare Dr. Smith for an independent research career by providing her with theoretical and knowledge-based training in the neurobiology of psychiatric disorders through formal course work in neuroscience, neurodevelopmental disorders and statistics, and through mentored research training in primary tissue culture, cell transplantation, behavioral analysis of transgenic mice, gene expression microarray experiments using the translating ribosome affinity purification (TRAP) method, and target validation experiments using in vivo electroporation of silencing RNA (siRNA) constructs. The long-term goal is to gain an understanding of the developmental events contributing to the maturation and survival of cortical interneurons during the critical period of their synaptic integration and cell maturation in the postnatal brain. Dr. Smith obtained her Ph.D. in genetics by performing candidate gene analysis of dopaminergic system genes in Attention Deficit Hyperactivity Disorder (ADHD). During her postdoctoral training at the Yale Child Study Center (YCSC), Dr. Smith participated in the T32 Neurobiology of Childhood Neuropsychiatric Disorders training program and has gained expertise in developmental neurobiology, anatomical analysis and behavioral characterization of transgenic mice while performing several studies to elucidate the role of fibroblast growth factor (Fgf) signaling in cortical development and behavior. Dr. Smith will continue her studies at YCSC, under the primary mentorship of Dr. Flora Vaccarino, with the assistance of a distinguished group of scientific advisors consisting of experts in psychiatry, neurobiology, and molecular genetics, who will provide guidance with her research design, performance of experiments, and data analysis. The diverse research environment at Yale offers numerous opportunities to learn from internationally recognized experts in the fields of psychiatry, neurobiology, and developmental biology and to attend seminars hosted by the various departments in neurobiology and psychiatric research. Dr. Smith will take advantage of these resources in order to attend courses, seminar series, journal clubs, and hands on training at core facilities. Dr. Smith will enhance these activities by attending scientific conferences and an intense workshop course in neuroscience methodologies. In the research proposal, Dr. Smith will utilize mice lacking the fibroblast growth factor receptor 1 (Fgfr1) gene, which exhibit a decrease in PV+ cortical inhibitory interneurons that is correlated with hyperactive behavior, in order to gain a better understanding of how the neuronal circuitry involved in behavioral inhibition is established. The loss of interneurons in Fgfr1 mutants occurs postnatally, when Fgfr1 is expressed in glial cells of the cortex, particularly astrocytes. Dr. Smith will investigate whether the astrocytes of Fgfr1 mutants are less capable of supporting the survival and maturation of cortical interneurons by in vitro co-culture, and by cell transplantation of interneurons into control and Fgfr1 mutant cerebral cortices. She will generate astrocyte specific mutations of Fgfr1 to test the hypothesis that Fgfr1 signaling in astrocytes is essential for proper maturation of inhibitory interneurons. Dr. Smith will utilize TRAP and microarray analysis to identify pathways that are disrupted by Fgfr1 mutations in astrocytes, and will test the effects of candidate genes upon interneuron maturation by in vivo electroporation of silencing RNAs. These studies will allow Dr. Smith to determine the role of Fgfr1 in establishing the proper proportion of PV+ interneurons in the cortex, and will shed light into the postnatal development of PV+ interneurons, a problem with direct relevance to schizophrenia and bipolar depression. Results from this research will inform future studies aimed at promoting the health and maturation cortical interneurons.

描述（由申请人提供）：gaba能抑制性中间神经元的缺失或功能障碍，特别是表达小白蛋白（PV）的中间神经元，与包括精神分裂症、双相抑郁症和妥瑞特综合征在内的严重精神疾病有关。PV+中间神经元调节兴奋性神经元的输出，对协调大脑节律很重要，因此兴奋性神经元可以同步放电-这些特性被认为对它们在认知和行为控制中的功能至关重要。本计划书描述了Karen M bbbbler Smith博士在导师指导下的职业发展培训期间的职业发展计划和研究目标。该提案的直接目标是通过神经科学、神经发育障碍和统计学方面的正式课程，以及在原代组织培养、细胞移植、转基因小鼠行为分析方面的指导研究培训，为史密斯博士提供精神疾病神经生物学方面的理论和知识培训，为她的独立研究生涯做好准备。使用翻译核糖体亲和纯化（TRAP）方法进行基因表达微阵列实验，以及使用体内电穿孔沉默RNA （siRNA）构建物进行靶标验证实验。长期目标是了解大脑皮层中间神经元在突触整合和细胞成熟的关键时期的成熟和存活的发育事件。Smith博士通过对注意缺陷多动障碍（ADHD）的多巴胺能系统基因进行候选基因分析获得了遗传学博士学位。在耶鲁儿童研究中心（YCSC）接受博士后培训期间，Smith博士参加了T32儿童神经精神疾病神经生物学培训项目，并获得了发育神经生物学、解剖分析和转基因小鼠行为表征方面的专业知识，同时进行了几项研究，阐明了成纤维细胞生长因子（Fgf）信号传导在皮层发育和行为中的作用。Smith博士将在YCSC继续她的研究，在Flora Vaccarino博士的主要指导下，在由精神病学，神经生物学和分子遗传学专家组成的杰出科学顾问小组的帮助下，他们将为她的研究设计，实验表现和数据分析提供指导。耶鲁大学多样化的研究环境提供了许多机会，可以向精神病学、神经生物学和发育生物学领域的国际知名专家学习，并参加由神经生物学和精神病学研究的各个部门主办的研讨会。史密斯博士将利用这些资源参加课程、系列研讨会、期刊俱乐部和核心设施的实践培训。史密斯博士将通过参加科学会议和神经科学方法论的密集研讨会课程来加强这些活动。在研究计划中，Smith博士将利用缺乏成纤维细胞生长因子受体1 （Fgfr1）基因的小鼠，这种小鼠表现出PV+皮质抑制性中间神经元的减少，这与过度活跃的行为有关，以便更好地了解参与行为抑制的神经元回路是如何建立的。Fgfr1突变体的中间神经元的丢失发生在出生后，当Fgfr1在皮层的胶质细胞，特别是星形胶质细胞中表达时。Smith博士将通过体外共培养和中间神经元细胞移植到对照和Fgfr1突变的大脑皮层中，研究Fgfr1突变体的星形胶质细胞是否支持皮层中间神经元存活和成熟的能力较差。她将产生星形胶质细胞特异性的Fgfr1突变，以验证星形胶质细胞中的Fgfr1信号对于抑制性中间神经元的适当成熟至关重要的假设。Smith博士将利用TRAP和微阵列分析来识别星形胶质细胞中被Fgfr1突变破坏的通路，并将通过沉默rna的体内电穿孔来测试候选基因对中间神经元成熟的影响。这些研究将使Smith博士能够确定Fgfr1在建立皮层中PV+中间神经元的适当比例中的作用，并将阐明PV+中间神经元的出生后发育，这是一个与精神分裂症和双相抑郁症直接相关的问题。本研究结果将为进一步研究促进皮层中间神经元的健康和成熟提供参考。