Mouse vestibular regeneration and function

小鼠前庭再生和功能

基本信息

批准号：
10528434
负责人：
Alan Gi-Lun Cheng
金额：
$ 58.73万
依托单位：
STANFORD UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-12-01 至 2023-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10528434
关键词：
Ablation Acceleration Adult Age Algorithmic Analysis Animals Bioinformatics Candidate Disease Gene Cell Survival Cell physiology Cells Chickens Cochlea Data Data Analyses Dimensions Disease Dizziness Electric Capacitance Electrophysiology (science)Epithelium Equilibrium Evoked Potentials Functional disorder Gene Expression Genes Genetic Goals Hair Cells Histologic Histology Human Impairment In Situ Hybridization Label Legal patent Mammals Maps Measurement Measures Mechanoreceptor Cell Methods Morphology Mus Natural regeneration Organ Physiology Property Recovery Recovery of Function Research Role Sensory Sensory Hair Supporting Cell Synapses Techniques Technology Testing Time Transgenic Mice Transgenic Organisms Utricle structure Vertigo Vestibular Hair Cells Work Zebrafish candidate identification cell regeneration design effective therapy experimental study gain of function gene discovery gene expression database genetic signature hair cell regeneration improved in vivo insight loss of function mechanotransduction mouse model nerve supply novel organ regeneration overexpression partial recovery prevent regeneration model repaired response restoration single-cell RNA sequencing spatiotemporal stem cells transcription factor transcriptome vesicular release

项目摘要

Abstract: Sensory hair cells are required for balance function. Vestibular hair cell degeneration causes balance dysfunction/hypofunction manifested as dizziness and vertigo. While the mammalian cochlea lacks the ability to regenerate lost hair cells, a limited degree of spontaneous regeneration occurs in the utricle, a vestibular organ detecting linear acceleration. Recent studies using fate-mapping techniques have pinpointed supporting cells as precursors of regenerated hair cells. However, it is not clear whether regenerated hair cells are fully functional and if organ function recovers. In preliminary experiments we have characterized hair cell degeneration and regeneration in the mature mouse utricle and also a loss followed by recovery of vestibular evoked potentials (VsEP) in vivo. The first aim of this proposal is to determine if increasing hair cell regeneration improves the recovery of vestibular function. Specifically, regenerated hair cells labeled via fate-mapping are probed via histology and electrophysiology to assess bundle morphology, mechanosensitvity, basolateral currents, and synaptic properties including vesicle release. In parallel, VsEP responses are measured and compared to histologic and electrophysiological measures. Next, by overexpressing Atoh1 via a transgenic approach, we will study the histology and electrophysiology of Atoh1-overexpressing hair cells and also the overall VsEP responses. In the second aim, we will determine if Atoh1 deletion prevents hair cell regeneration and the recovery of VsEP responses. In parallel, fate-mapped, surviving hair cells will be examined for possible repair via histology and electrophysiology. To gain an unbiased insight into the genetic signature of hair cell progenitors and surviving hair cells, the third aim is designed to examine the damaged mature mouse utricle using single cell RNA sequencing technologies. Here the first goal is to discover the genetic landscape of hair cell progenitors and surviving hair cells in the damaged utricle. Secondly, we will examine the gene expression of the undamaged and damaged utricle after Atoh1 overexpression. Lastly, we will use bioinformatic approaches to delineate the trajectory of the spontaneous and Atoh1-enhaced supporting cell-hair cell transition and validate this histologically. In summary, we will apply state-of-the art technologies (vestibular physiology, hair cell physiology, single cell RNA-seq, bioinformatic strategies) to study vestibular hair cell regeneration in transgenic mouse models. We have assembled a team of experts who have worked together to collect promising preliminary data. At the end of this 5-year proposal, we will have 1) determined the relationship between hair cell regeneration and functional recovery and 2) revealed and temporally ordered novel genes during mammalian hair cell regeneration.

抽象的：平衡功能需要感觉毛细胞。前庭毛细胞变性会导致平衡功能障碍/功能障碍表现为头晕和眩晕。而哺乳动物的耳蜗缺乏能力再生毛细胞的再生，有限程度的自发再生发生在乌尔特里斯特，前庭器官检测线性加速度。使用命运映射技术的最新研究已将支持细胞指定为再生毛细胞的前体。但是，尚不清楚再生的毛细胞是否完全功能如果器官功能恢复。在初步实验中，我们表征了毛细胞变性和成熟的小鼠尿囊的再生，也损失，然后恢复前庭诱发电位（VSE）在体内。该提案的第一个目的是确定增加毛细胞再生是否有所改善恢复前庭功能。具体而言，通过命运映射标记的再生毛细胞通过组织学和电生理学，以评估束形态学，机械感应，基底外侧电流和突触特性，包括囊泡释放。同时，测量了VSEP响应，并将其与组织学和电生理测度。接下来，通过通过转基因方法过表达ATOH1，我们将研究ATOH1过表达毛细胞的组织学和电生理学以及总体VSEP 回答。在第二个目标中，我们将确定ATOH1缺失是否阻止了毛细胞再生和恢复 VSEP响应。同时，将检查命运图，存活的毛细胞，以通过组织学修复可能修复和电生理学。为了对毛细胞祖细胞的遗传特征和幸存毛细胞是第三个目标，旨在使用单细胞RNA检查受损的成熟小鼠尿。测序技术。在这里，第一个目标是发现毛细胞祖细胞和在受损的尿液中存活的毛细胞。其次，我们将检查未损坏的基因表达并在AtoH1过表达后损坏的尿液。最后，我们将使用生物信息学方法来描述自发和ATOH1启用的轨迹支撑细胞毛细胞过渡并验证这一点组织学上。总而言之，我们将采用最先进的技术（前庭生理学，毛细胞生理学，单细胞RNA-seq，生物信息学策略）研究转基因小鼠中的前庭毛细胞再生型号。我们组建了一个专家团队，他们共同努力收集有希望的初步数据。在这个5年提案的结尾，我们将有1）确定毛细胞再生之间的关系和功能恢复和2）在哺乳动物毛细胞期间揭示和暂时有序的新基因再生。