Transcriptional regulation of hair-cell progenitors in the zebrafish lateral line

斑马鱼侧线毛细胞祖细胞的转录调控

• 批准号: 9305702
• 负责人: Aaron Steiner
• 金额: $ 37.85万
• 依托单位: PACE UNIVERSITY NEW YORK
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-04-01 至 2021-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9305702
• 关键词: Ablation; American; Amphibia; Animals; Anterior; Automobile Driving; Back; Biological Assay; Biological Models; Biology; Birds; Candidate Disease Gene; Cell Death; Cell Lineage; Cell Proliferation; Cells; Chemicals; Cochlear Implants; Comprehension; Data Set; Deterioration; Development; Down-Regulation; Ear; Event; FAT gene; Family; Family member; Fatty acid glycerol esters; Fishes; Flow Cytometry; Fostering; Future; Gene Expression Profiling; Gene Family; Gene Transfer Techniques; Genealogy; Genes; Goals; Hair Cells; Hearing; Hearing Aids; Human; Individual; Injection of therapeutic agent; Integral Membrane Protein; Label; Labyrinth; Life; Mammals; Mediating; Messenger RNA; Microarray Analysis; Molecular; Natural regeneration; Organ; Organ Size; Pathway interactions; Pattern; Population; Positioning Attribute; Procedures; Process; Public Health; Recovery; Reptiles; Research; Role; Sensory; Signal Transduction; Stem cells; System; Testing; Time; Transcript; Transcriptional Regulation; Transgenes; Transgenic Organisms; United States; Vertebrates; Water; Zebrafish; alternative treatment; base; cancer cell; experimental study; gain of function; hair cell regeneration; hearing impairment; interest; knock-down; lateral line; loss of function; member; mutant; neuromast; ototoxicity; overexpression; paralogous gene; progenitor; reconstitution; regenerative; regenerative therapy; response; sensory system; therapy development; tool

Project Summary Hearing loss is a potentially debilitating condition that afflicts to varying extents more than 30 million individuals in the United States. Commonly caused by destruction of the mechanosensory hair cells in the inner ear, most hearing loss in humans is permanent because these cells are not naturally replenished. Unlike those in the human ear, hair cells of non-mammalian vertebrates including fishes, amphibians, and reptiles including birds can regenerate throughout life. Despite extensive research, the molecular and cellular bases of this difference are not yet well understood. Because a more complete comprehension of hair-cell regeneration in non-mammalian model systems will foster the development of regenerative therapies for hearing loss, our current research focuses on elucidating the mechanism of hair-cell regeneration in one such system, the zebrafish lateral line. The lateral line comprises an array of superficially located hair cell-containing organs called neuromasts that detect water displacement. We have devised procedures for transgenically labeling and isolating the putative progenitors—called mantle cells—that are thought to give rise to hair cells in neuromasts during regeneration. Microarray analysis has revealed numerous genes that are highly and specifically expressed in mantle cells, as well as genes whose expression in these cells is modulated in response to ototoxic insult. The resulting data set places us in a unique position to determine which molecular pathways activate or repress the proliferation of progenitors and the subsequent replacement of hair cells. Among the top candidate genes for controlling progenitor proliferation are those encoding the transmembrane protein Tspan1 and the protocadherins Fat1a and Fat1b, which are highly enriched in mantle cells and are down- regulated during regeneration. We propose to use loss-of-function (antisense-mediated knockdown and existing mutant lines) as well as gain-of-function (mRNA-mediated overexpression and misexpression by means of an inducible transgenic system) to test the functions of Tspan1, Fat1a, and Fat1b in hair-cell development and regeneration. Through these studies we hope to uncover previously unappreciated molecular mechanisms governing hair-cell regeneration, which may contribute to the future development of therapies for hearing loss.

项目摘要 听力损失是一种潜在的使人衰弱的状况，影响了超过3000万 在美国的个人。通常是由机械学毛细胞破坏引起的 内耳，人类的大多数听力损失都是永久的，因为这些细胞不是自然的 更换。与人耳中的那些不同，非哺乳动物脊椎动物的毛细胞在内 两栖动物和包括鸟类在内的爬行动物可以在一生中再生。尽管进行了广泛的研究， 该差异的分子和细胞碱基尚不清楚。因为更完整 对非哺乳动物模型系统中发型细胞再生的理解将促进发展 关于听力损失的再生疗法，我们目前的研究重点是阐明 在一个这样的系统中，发胶再生是斑马鱼的侧线。横向线包含一个数组 含毛细胞细胞的器官，称为神经瘤，可检测水位的位移。 我们已经设计了翻译标记和隔离假定祖细胞的程序 - 地幔细胞 - 被认为会在再生过程中引起神经瘤的毛细胞。微阵列 分析揭示了许多在地幔细胞中高度和特异性表达的基因 作为对耳毒性侮辱的基因，其在这些细胞中的表达受到调节。结果数据 将我们设置为独特的位置，以确定哪种分子途径激活或反映 祖细胞的增殖和随后的毛细胞的替代。在顶级候选人中 用于控制祖细胞增殖的基因是编码跨膜蛋白TSPAN1的基因 以及高度富含地幔细胞的protastadherins fat1a和fat1b 在再生过程中受到调节。我们建议使用功能丧失（反义介导的敲低 和现有的突变线）以及功能获得（mRNA介导的过表达和 通过诱导的转基因系统的misexpression测试TSPAN1，FAT1A和 发胶发育和再生中的FAT1B。通过这些研究，我们希望以前发现 主管收发再生的未批准的分子机制，这可能有助于 未来开发听力损失的疗法。

项目成果

Confocal Microscope-Based Laser Ablation and Regeneration Assay in Zebrafish Interneuromast Cells.

• DOI: 10.3791/60966
• 发表时间: 2020-05-20
• 期刊: JOVE-JOURNAL OF VISUALIZED EXPERIMENTS
• 影响因子: 1.2
• 作者: Volpe, Bryan A.;Fotino, Teresa H.;Steiner, Aaron B.
• 通讯作者: Steiner, Aaron B.