Neural defects in zebrafish auditory/vestibular mutants

斑马鱼听觉/前庭突变体的神经缺陷

基本信息

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
项目类别：
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

项目摘要

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央前庭疾病。此外，听力损失或耳鸣可能具有中心起源。尽管中心盛行患者听觉/前庭系统的缺陷，我们对中枢功能障碍的理解缺乏脊椎动物中的分子或细胞水平。在这里，我们建议描述小说具有中央听觉/前庭缺陷的斑马鱼突变体类别研究的研究领域。该提议重点介绍了两个突变体：Raumschiff和Starliner，从化学诱变筛选中分离出来，以进行听力和平衡缺陷。与我们先前表征的突变体不同，Raumschiff和Starliner突变体正常前庭诱导的眼动运动，尽管存在明显的平衡缺陷，而游泳或休息。另外，两个突变体中都存在听力缺陷。我们有在两个基因中鉴定出突变：在Starliner突变体中，分裂末端（SPEN）基因具有A 废话突变和raumschiff中，我们确定了液泡蛋白的错义突变排序4A（VPS4A）。突变体和兄弟姐妹转录本的rnaseq anaylsis表明两个突变体在后脑和中脑区域对基因表达的震撼人心发育和大脑形态是正常的。这些结果表明缺陷是本质上的功能，可能涉及电路水平或突触变化。获得更好的了解中心缺陷，我们将利用整个鱼表达的成像相关的转基因标记，并使用新开发的方法用于细胞的脑部成像对听觉和前庭刺激的反应。这些实验将集中于区域或细胞基因表达不正常的类型最普遍。我们的学习将增强我们对涉及中心功能的基因和调节网络的理解。