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: 7541457
• 负责人: Helen Lai
• 金额: $ 5.36万
• 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/7541457
• 关键词: Alzheimer' s Disease; BHLH Protein; Binding Sites; Bioinformatics; Biological Assay; 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; Reporter; Reporter Genes; Research; Specific qualifier value; System; Testing; To specify; Transcript; Transgenic Mice; Work; Yang; cell type; chromatin immunoprecipitation; feeding; granule cell; mutant; overexpression; research study; spinal cord and brain injury; 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基本螺旋-环-螺旋（bHLH）转录因子 Atoh1 控制神经元细胞类型规范的调节机制。由于缺乏已知的 Atoh1 靶标，Atoh1 编码背神经管中的 d!1 神经元和小脑中的颗粒细胞的方式仍不清楚。这项研究将重点鉴定 Atoh1 靶标转录因子，并揭示指定神经元细胞类型的 Atoh1 顺式调控代码。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规范的分子机制，可能有助于确定神经退行性疾病、脑和脊髓损伤以及神经元细胞衍生癌症的治疗靶点。具体目标是： 1．确定 Atoh1 靶点并确定这些靶点的功能上重要的调控区域。微阵列实验已经鉴定出 Atoh1 谱系细胞中富含的转录本。 Atoh1 的下游靶标将从转录因子候选列表中确定，并在小鸡和转基因小鼠试验中测试报告基因的正确表达，并测试其调控区域。 Atoh的直接靶标将通过染色质免疫沉淀(ChIP)方法来鉴定。2．确定 Atoh1 指定神经元的顺式调节代码。将探测 Atoh1 直接靶标的调控区域中的 Atoh1 共有结合位点，并使用生物信息学分析检查 Atoh1 靶标的调控区域中的常见转录因子结合位点或 DNA 序列基序。候选监管代码将通过小鸡突变或转基因小鼠报告基因检测进行功能测试。监管代码将用于通过目标 1 和 2 的迭代循环进一步确定 Atoh1 的目标。 我试图了解神经元在最基本的层面上是如何形成的。我的研究可能会帮助我们了解如何通过治疗方法恢复患有大脑和脊髓损伤、阿尔茨海默氏症或帕金森氏症等神经退行性疾病或儿童脑肿瘤患者的工作神经。