Genome Wide Identification of PTF1-J Targets in Dorsal Neural Tube

背神经管 PTF1-J 靶标的全基因组鉴定

• 批准号: 7928766
• 负责人: Jane E Johnson
• 金额: $ 19.43万
• 依托单位: UT SOUTHWESTERN MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-15 至 2011-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7928766
• 关键词: Autistic Disorder; Binding; Binding Sites; Bioinformatics; Biological Assay; Boxing; Cell Line; Cells; Cerebellum; Characteristics; Chromatin; Complex; Consensus; Consensus Sequence; DNA; DNA Sequence; Development; Developmental Process; Disease; Dorsal; E protein; Embryo; Embryonic Development; Epilepsy; Equilibrium; Exploratory/Developmental Grant; Funding; Gene Expression Profile; Gene Targeting; Generations; Genetic Transcription; Goals; Hyperalgesia; In Vitro; Molecular; Molecular Profiling; Nerve Degeneration; Nervous System Physiology; Neural tube; Neuraxis; Neuronal Differentiation; Neurons; Nucleic Acid Regulatory Sequences; Pathway interactions; Phenotype; Protocols documentation; Relative (related person); Reporter; Retina; Signal Transduction; Site; Spinal Cord; Staging; Stem cells; Testing; Vertebrates; Work; chromatin immunoprecipitation; embryo tissue; excitatory neuron; genome wide association study; high risk; in vivo; inhibitory neuron; insight; mutant; nerve stem cell; nervous system disorder; neurodevelopment; notch protein; public health relevance; relating to nervous system; research study; transcription factor

DESCRIPTION (provided by applicant): Ptf1a is required for GABAergic inhibitory neurons in the dorsal spinal cord, cerebellum, and retina. In the absence of Ptf1a, neural progenitor cells fail to generate inhibitory neurons and aberrantly assume an excitatory neuronal phenotype. Ptf1a is a class II bHLH factor that can heterodimerize with class I bHLH factors (E-proteins) to bind E-box DNA consensus sequences in vitro. However, the in vivo function of Ptf1a is in a unique transcription factor complex, called PTF1-J, that contains Ptf1a and an E-protein, plus a third factor Rbpj. This complex binds a bipartite DNA sequence containing the E-box plus a TC- box. Rbpj has a distinct function as the transcriptional effector of Notch signaling. Notch-Rbpj signaling inhibits neuronal differentiation, while Ptf1a-Rbpj functions in neuronal subtype specification. The proposal is to use in vivo chromatin immunoprecipitation combined with massive parallel sequencing platforms (ChIP-seq) to delineate the function of Ptf1a by genome wide identification of its transcription targets. ChIP-seq identified target genes of Ptf1a in vivo will be validated and characterized by combining results from expression profiling of Ptf1a cells in the dorsal neural tube, evaluating changes in expression in Ptf1a mutants, and by testing target regulatory regions in reporter assays in chick neural tube with and without exogenous Ptf1a. Identifying targets of Ptf1a will provide molecular insight into developmental processes controlling the balance of inhibitory and excitatory neurons. PUBLIC HEALTH RELEVANCE: Normal nervous system function requires the correct balance of inhibitory and excitatory neurons. Alterations in this balance are thought to underlie diverse neurological disorders from epilepsy to autism to hyperalgesia. Successful completion of these aims will substantially advance efforts to define transcriptional networks regulating the balance of inhibitory and excitatory neuronal specification. This is a unique opportunity to combine multiple technical advances to begin to tackle the challenging goal of defining the molecular rationale controlling the generation of neuronal diversity.

描述（由申请人提供）：Ptf 1a是背侧脊髓、小脑和视网膜中GABA能抑制性神经元所必需的。在没有Ptf 1a的情况下，神经祖细胞不能产生抑制性神经元，并且异常地呈现兴奋性神经元表型。Ptf 1a是一种II类bHLH因子，可与I类bHLH因子（E蛋白）异源二聚化，以在体外结合E-box DNA共有序列。然而，Ptf 1a的体内功能是在一个独特的转录因子复合物中，称为PTF 1-J，它含有Ptf 1a和E蛋白，加上第三个因子Rbpj。该复合物结合含有E盒加TC盒的二分DNA序列。Rbpj具有作为Notch信号传导的转录效应子的独特功能。Notch-Rbpj信号传导抑制神经元分化，而Ptf 1a-Rbpj在神经元亚型特化中起作用。该提案是使用体内染色质免疫沉淀结合大规模并行测序平台（ChIP-seq），通过全基因组范围内识别其转录靶点来描绘Ptf 1a的功能。将通过结合背神经管中Ptf 1a细胞的表达谱分析结果、评价Ptf 1a突变体中的表达变化以及在有和无外源Ptf 1a的鸡神经管中检测报告基因测定中的靶调控区，验证和表征ChIP-seq体内鉴定的Ptf 1a靶基因。确定Ptf 1a的靶点将为控制抑制性和兴奋性神经元平衡的发育过程提供分子见解。 公共卫生相关性：正常的神经系统功能需要抑制性和兴奋性神经元的正确平衡。这种平衡的改变被认为是从癫痫到自闭症再到痛觉过敏的各种神经系统疾病的基础。这些目标的成功完成将大大推进努力，以确定调节抑制性和兴奋性神经元规范的平衡的转录网络。这是一个独特的机会，联合收割机多种技术进步，开始解决具有挑战性的目标，定义控制神经元多样性的产生的分子原理。