Neural defects in zebrafish auditory/vestibular mutants

斑马鱼听觉/前庭突变体的神经缺陷

• 批准号: 10412441
• 负责人: Teresa A Nicolson
• 金额: $ 47.42万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-01 至 2022-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10412441
• 关键词: Acoustics; Affect; Afferent Neurons; Anatomy; Animals; Area; Auditory; Auditory system; Behavioral; Biogenesis; Biological Process; Biology; Brain; Calcium; Chemicals; Cues; Data; Data Set; Defect; Development; Disease; Dizziness; Down-Regulation; ENG gene; Epigenetic Process; Equilibrium; Eye Movements; Failure; Fishes; Functional disorder; Gene Expression; Gene Expression Profile; Gene Expression Regulation; Gene Targeting; Generations; Genes; Genetic; Genetic Transcription; German population; Goals; Hair Cells; Hearing; Human; Image; Imaging Device; Impairment; In Situ Hybridization; Label; Labyrinth; Larva; Lesion; Link; Location; Methods; Midbrain structure; Missense Mutation; Molecular; Molecular Analysis; Mutagenesis; Mutation; Nature; Neuraxis; Neurologic; Neurons; Nonsense Mutation; Optics; Patients; Peripheral; Phenotype; Physiological; Physiology; Population; Posture; Prevalence; Proteins; Proxy; Reflex action; Research; Resolution; Rest; Siblings; Signal Transduction; Stimulus; Swimming; Synapses; System; Testing; Tinnitus; Transcript; Untranslated RNA; Up-Regulation; Vacuolar Protein Sorting; Vertebrates; Vertigo; Zebrafish; attenuation; base; behavioral phenotyping; behavioral response; brain morphology; cell type; cellular imaging; clinical heterogeneity; exosome; experimental study; forward genetics; genetic approach; hearing impairment; hearing screening; hindbrain; insight; macula; marker transgenes; mechanotransduction; mutant; novel; relating to nervous system; response; transcription factor; transcriptome sequencing; transcriptomics

PROJECT SUMMARY Approximately one quarter of patients with vertigo or dizziness have central vestibular disorders. In addition, hearing loss or tinnitus can have central origins. Despite the prevalence of central deficits in the auditory/vestibular system in patients, our understanding of central dysfunction at the molecular or cellular level in vertebrates is lacking. Here, we propose to characterize a novel class of zebrafish mutants that have central auditory/vestibular deficits to gain insights into this understudied area of research. This proposal focuses on two mutants: raumschiff and starliner, which were isolated from chemical mutagenesis screens for hearing and balance defects. Unlike our previously characterized mutants, raumschiff and starliner mutants have normal vestibular induced eye movements despite presenting with an obvious balance defect while swimming or at rest. In addition, a defect in hearing is present in both mutants. We have identified mutations in two genes: in starliner mutants, the split ends (spen) gene harbors a nonsense mutation and in raumschiff we identified a missense mutation in vacuolar protein sorting 4a (vps4a). RNAseq anaylsis of mutant and sibling transcripts indicate that both mutants have striking misregulation of gene expression in the hindbrain and midbrain regions, yet their development and gross brain morphology is normal. These results suggest that the defects are functional in nature and may involve circuit level or synaptic changes. To gain a better understanding of the central defects, we will take advantage of imaging whole fish expressing relevant transgene markers and use newly developed methods for brain-wide imaging of cellular responses to auditory and vestibular stimuli. These experiments will focus on the regions or cell types where misregulation of gene expression is most prevalent. Collectively our studies will enhance our understanding of the genes and regulatory networks involved in central function.

项目摘要 大约四分之一的眩晕或头晕患者患有中枢性前庭障碍。 此外，听力损失或耳鸣可以有中央起源。尽管中央 缺陷的听觉/前庭系统的患者，我们的理解中枢功能障碍， 在脊椎动物中缺乏分子或细胞水平。在这里，我们建议将一部小说 一类具有中枢听觉/前庭缺陷的斑马鱼突变体， 未被充分研究的领域。这个提议集中在两个突变体上：raumschiff和starliner， 其是从针对听力和平衡缺陷的化学诱变筛选中分离的。 不像我们以前描述的突变体，raumschiff和starliner突变体有正常的 尽管存在明显的平衡缺陷，但前庭诱导的眼球运动， 游泳或休息。此外，两种突变体都存在听力缺陷。我们有 在两个基因中发现了突变：在starliner突变体中，分裂末端（spen）基因携带一个 在raumschiff中，我们发现了一个空泡蛋白的错义突变， 排序4a（vps 4a）。对突变体和同胞转录物的RNAseq分析表明，两种突变体均 在后脑和中脑区域有显著的基因表达失调， 发育和大体脑形态正常。这些结果表明， 本质上是功能性的，并且可能涉及电路水平或突触变化。更好地 了解中心缺陷，我们将利用成像全鱼表达 相关的转基因标记，并使用新开发的方法进行细胞的全脑成像， 对听觉和前庭刺激的反应。这些实验将集中在区域或细胞 基因表达失调最为普遍的类型。我们的研究将 增强我们对参与中枢功能的基因和调控网络的理解。