Gene Expression Profiling of the Development Inner Ear

内耳发育的基因表达谱

• 批准号: 6618532
• 负责人: Michael Lovett
• 金额: $ 32.29万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-04-01 至 2007-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6618532
• 关键词: complementary DNA; developmental neurobiology; ectoderm; embryonic stem cell; gene expression; gene expression profiling; gene induction /repression; genetic library; histogenesis; human genetic material tag; in situ hybridization; laboratory mouse; labyrinth; laser capture microdissection; mesoderm; microarray technology; nucleic acid purification; phenotype; polymerase chain reaction; transcription factor

DESCRIPTION (provided by applicant): We shall employ micro-cDNA methods, gene chips and custom oligonucleotide microrarrays to investigate which genes are specifically up or down regulated in the early stages of mammalian inner ear development the progenitors of the inner ear appear as an ectodermal thickening at about day 8 of mouse development gradually this structure invaginates and the otocyst/otic vesicle is formed which by day 15 postcoitum holds most of the complex cellular machinery of the inner ear we intend to conduct a detailed gene chip analysis of this time course in the mouse we shall also employ a custom transcription factor microarray to measure expression changes in these important regulatory molecules early inductive mechanisms will be investigated by comparing gene expression in mesoderm, hind brain and the ectodermal thickening that precedes actual otic placode development in addition to these relatively conventional analyses, we shall explore innovative cdna target depletion strategies designed to measure changes in much lower abundance transcripts our third aim is to develop a custom oligonucleotide microarray of important genes identified using the gene chip data and by interrogation of a large microarray database we shall use this to reanalyze our mouse time courses and for a more limited parallel study of microdissected human embryonic otic structures to confirm the relevance of our observations to human ear development we shall conduct confirmatory assays to validate our observations from expression arrays finally, we intend to apply the knowledge gained from normal otic development to an investigation of expression profiles in mouse embryonic stem cell lines preliminary data indicate that is cells can be initiated into what may be a "otic" pathway we shall investigate the gene expression profiles in these clones of cells and also apply the normal otic expression profiles to "directed differentiation" experiments in the es system we expect that the type of gene expression profiles we shall build for the developing inner ear will highlight candidate genes for hearing loss, that they will pinpoint at least some of the players in a sequential genetic pathway leading to auditory hair cell differentiation and innervation (as just two examples), and that they will prove useful in the intelligent design of differentiation strategies for embryonic stem cells for future replacement therapies.

描述（由申请人提供）：我们应采用微cDNA方法，基因芯片和自定义寡核苷酸微阵列来调查哪些基因在内耳的早期阶段被调节或向下调节，内耳的早期阶段哺乳动物的哺乳动物的祖细胞发育的哺乳动物的祖细胞的内耳的祖细胞看起来像逐渐散发出这种结构的近年三天的外形增长，即逐渐散发出的结构。涂料后持有内耳的大部分复杂的细胞机械，我们打算对鼠标中的这一时间过程进行详细的基因切解分析，我们还将采用自定义的转录因子微阵列来衡量这些重要的调节分子中早期电感性机制中的表达变化，将通过比较这些脑中的基因表达，以使其在中间和脑demente液中进行比较，从而在中度和脑dementer脑中进行比较，从而在中度和脑中的进化，以实现对进一步的良性，以实现对进一步的进度，以实现进一步的进展，以实现进一步的进展，以实现进一步的进展，以实现进一步的进展。传统分析，我们将探索创新的cDNA目标耗竭策略，旨在衡量较低的丰度转录本的变化我们的第三个目标是开发一种自定义的重要基因的自定义寡核苷酸微阵列，该基因使用基因芯片数据识别出来的重要基因，并通过询问我们的大型微阵列数据库，我们将使用此构造的构造，我们将更加限制地研究了鼠标，我们将更多地研究鼠标。为了确认我们观察到的人耳朵发育的相关性，我们应进行验证性测定，以验证我们从表达阵列中的观察结果最终，我们打算将从正常的动态发育中获得的知识应用于对小鼠胚胎干细胞系中表达谱的研究的调查，初步数据表明，细胞可以将其启动到可能是“ OIT的表达”中，我们还将在这些cloone中启动这些closiles，我们还将调查这些closiles的表达。差异化“在ES系统中进行的实验我们期望我们将为发展中的内耳所建立的基因表达谱类型将突出候选基因的听力损失，它们将在顺序的遗传途径中至少确定一些参与者，从而导致听觉毛细胞分化和神经支配（仅仅是两个例子），并且在两个示例中可以证明具有智能设计的启动型细胞，以换取型号的启动策略。