Transcriptional Regulatory Control of Neuronal Cell Type Specification

神经元细胞类型规范的转录调控

• 批准号: 7406319
• 负责人: Helen Lai
• 金额: $ 5.15万
• 依托单位: UT SOUTHWESTERN MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-12-15 至 2010-12-14
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7406319
• 关键词: Alzheimer' s Disease; BHLH Protein; Binding Sites; Bioinformatics; Biological Assay; Brain; Cell Lineage; Cells; Cerebellum; Childhood Brain Neoplasm; Code; Consensus; DNA Sequence; Data Set; Development; Dorsal; Experimental Designs; Genetic; Goals; Helix-Turn-Helix Motifs; In Situ Hybridization; Interneurons; Knockout Mice; Knowledge; Laboratories; Lead; Malignant Neoplasms; Methods; Molecular; Mus; Mutation; Nerve; Neural tube; Neurodegenerative Disorders; Neurons; Nucleic Acid Regulatory Sequences; Parkinson Disease; Population; Protein Overexpression; Reporter; Reporter Genes; Research; Specific qualifier value; Spinal cord injury; System; Testing; To specify; Transcript; Transgenic Mice; Work; Yang; cell type; chromatin immunoprecipitation; feeding; granule cell; mutant; research study; therapeutic target; transcription factor

DESCRIPTION (provided by applicant): The goal of the proposed research is to understand the regulatory mechanisms that govern neuronal cell type specification through the basic helix-loop-helix (bHLH) transcription factor, Atoh1. How d!1 neurons in the dorsal neural tube and granule cells in the cerebellum are encoded by Atoh1 remains undefined due to the lack of known Atoh1 targets. This study will focus on identifying transcription factors that are Atoh1 targets and uncover the Atoh1 cis-regulatory code that specifies neuronal cell types. The long term goal is to understand the molecular mechanisms of how neuronal bHLH transcription factors regulate neuronal cell type specification possibly leading to the identification of therapeutic targets for neurodegenerative diseases, brain and spinal cord injuries, and neuronal cell-derived cancers. The specific aims are:1. Identify Atoh1 targets and determine functionally important regulatory regions of these targets. Microarray experiments have identified transcripts enriched in Atoh1-lineage cells. Downstream targets of Atoh1 will be identified from a candidate list of transcription factors and their regulatory regions tested for proper expression of a reporter gene in chick and transgenic mouse assays. Direct targets of Atoh will be identified by chromatin immunoprecipitation (ChIP) approaches.2. Determine a cis-regulatory code by which Atoh1 specifies neurons. Regulatory regions of direct Atoh1 targets will be probed for an Atoh1 consensus binding site and regulatory regions of Atoh1 targets will be examined for common transcription factor binding sites or DNA sequence motifs using bioinformatics analyses. Candidate regulatory codes will be functionally tested by mutation in the chick or transgenic mouse reporter assays. Regulatory codes will be used to further identify targets of Atoh1 through iterative cycles of Aims 1 and 2. I am trying to understand how neurons form at their most basic level. My research may lead to discoveries in understanding how we can therapeutically restore working nerves in people with brain and spinal cord injuries, neurodegenerative diseases such as Alzheimer's or Parkinson's disease, or pediatric brain tumors.

描述（由申请人提供）：拟议研究的目标是了解通过碱性螺旋-环-螺旋（bHLH）转录因子Atoh 1控制神经元细胞类型特化的调控机制。怎么会这样！1背神经管中的神经元和小脑中的颗粒细胞由Atoh 1编码，由于缺乏已知的Atoh 1靶点，因此尚未确定。这项研究将集中在确定Atoh 1的目标转录因子，并揭示Atoh 1顺式调节代码，指定神经元细胞类型。长期目标是了解神经元bHLH转录因子如何调节神经元细胞类型特化的分子机制，可能导致神经退行性疾病、脑和脊髓损伤以及神经元细胞源性癌症的治疗靶点的鉴定。具体目标是：1.确定Atoh 1目标，并确定这些目标的功能重要的调控区域。微阵列实验已经鉴定了Atoh 1谱系细胞中富集的转录物。Atoh 1的下游靶标将从候选转录因子列表中鉴定，并在鸡和转基因小鼠试验中检测其调节区是否正确表达报告基因。Atoh的直接靶点将通过染色质免疫沉淀（ChIP）方法鉴定.确定Atoh 1指定神经元的顺式调节密码。使用生物信息学分析，探测直接Atoh 1靶标的调控区中的Atoh 1共有结合位点，并检查Atoh 1靶标的调控区中的常见转录因子结合位点或DNA序列基序。将通过鸡或转基因小鼠报告基因试验中的突变对候选调控代码进行功能检测。监管代码将用于通过目标1和2的迭代循环进一步识别Atoh 1的靶标。 我试图了解神经元在最基本的水平上是如何形成的。我的研究可能会导致发现，了解我们如何在治疗上恢复大脑和脊髓损伤，神经退行性疾病，如阿尔茨海默氏症或帕金森氏症，或小儿脑肿瘤患者的工作神经。