Characterizing hair cell regeneration in the larval zebrafish inner ear

斑马鱼幼虫内耳毛细胞再生的特征

• 批准号: 10604748
• 负责人: Marielle Beaulieu
• 金额: $ 4.68万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-16 至 2024-09-15
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10604748
• 关键词: Ablation; Amphibia; Animals; Auditory; Biological Models; Birds; Cell Nucleus; Cell Proliferation; Cell division; Cells; Cellular Morphology; Cessation of life; Clustered Regularly Interspaced Short Palindromic Repeats; Cochlea; Data; Data Set; Development; Ear; Embryonic Development; Fishes; Fluorescent in Situ Hybridization; Functional disorder; Future; Genetic; Genetic Techniques; Genetic Transcription; Growth; Hair Cells; Human; Image; Knowledge; Label; Labyrinth; Lead; Life; Location; Mammals; Mediating; Methods; Morphology; Natural regeneration; Neonatal; Neuroglia; Organ; Organism; Population; Proliferating; Regenerative capacity; Role; Sensory; Sensory Hair; Study models; Support Groups; Supporting Cell; System; Transgenic Organisms; Vertebrates; Work; Zebrafish; cell growth; cranium; deafness; experimental study; hair cell regeneration; in vivo; in vivo Model; in vivo imaging; in vivo regeneration; lateral line; molecular marker; mutant; novel; organ growth; organ regeneration; regeneration potential; regenerative; sensory system; single cell analysis; single-cell RNA sequencing; stem cells; tool; transdifferentiation; water flow; zebrafish development

PROJECT SUMMARY Death of sensory hair cells in the inner ear is a common cause of auditory and vestibular dysfunction in humans. This damage is permanent due to the inability of mammals to effectively regenerate inner ear sensory patches beyond the neonatal stage. In contrast, non-mammalian vertebrate species such as birds and fish can regenerate hair cells throughout life. This discrepancy in regeneration capacity between mammals and non- mammalian vertebrates is not well understood. In species that can regenerate hair cells, the nonsensory support cells that surround hair cells can act as hair cell progenitors. In vivo studies of inner ear hair cell regeneration and support cells are limited by the inner ear’s location within the bony skull of mammals and birds. As a result, the zebrafish lateral line, an external sensory system for detecting changes in water flow, has been developed as an alternative model for studying support cells and hair cell regeneration in living animals. However, the degree to which the mechanisms of regeneration in the lateral line are similar to those of the ear is unclear. The zebrafish inner ear is an understudied system that has the potential to bridge the gap between mammalian inner ear and lateral line studies. This project examines hair cell regeneration in the zebrafish inner ear during the larval stage, at which point the sensory organs become fully functional and remain accessible for in vivo imaging. We aim to identify hair cell precursors in the zebrafish inner ear and determine when and how they give rise to new hair cells during regeneration. In the inner ear of birds and mammals, support cells regenerate hair cells by proliferating and differentiating or by direct transdifferentiation. In the zebrafish lateral line, only the former mechanism is used. In Aim 1 of this proposal, we will use a novel method of hair cell ablation to determine the timecourse of regeneration and whether cell proliferation, transdifferentiation, or both are used in the zebrafish inner ear. Hair cells in the lateral line are regenerated primarily by a spatially and functionally distinct group of support cells. Whether a similar support cell subtype exists in the zebrafish inner ear is unknown. In Aim 2, we will validate putative inner ear support cell subtypes identified through single-cell RNA-seq analysis. We will then generate transgenic lines labeling support cell subtypes and use fate mapping to identify hair cell precursors. Together these experiments will characterize the timing and mechanism of hair cell regeneration and the diversity of support cell subtypes in the zebrafish inner ear. This work will help to establish the zebrafish inner ear as an outstanding in vivo model system for future in-depth studies of support cells and their roles in conserved mechanisms of regeneration. Expanding our knowledge of the mechanisms of inner ear hair cell regeneration in animals such as the zebrafish will ultimately lead to the advancement of treatments for auditory and vestibular decline in humans.

项目摘要 内耳感觉毛细胞的死亡是听觉和前庭功能障碍的常见原因， 人类这种损伤是永久性的，因为哺乳动物无法有效地再生内耳感觉。 超过新生儿阶段的斑块。相比之下，非哺乳类脊椎动物物种，如鸟类和鱼类， 再生毛细胞在整个生命。哺乳动物和非哺乳动物在再生能力上的差异 哺乳类脊椎动物还不太了解。在能够再生毛细胞的物种中， 毛细胞周围的支持细胞可以作为毛细胞祖细胞。内耳毛细胞的体内研究 再生和支持细胞受到内耳在哺乳动物骨头骨内的位置的限制， 鸟因此，斑马鱼的侧线，一种检测水流变化的外部感觉系统， 已被开发为研究支持细胞和毛细胞再生的替代模型， 动物然而，在何种程度上的再生机制在侧线是相似的， 耳朵不清楚。斑马鱼的内耳是一个未充分研究的系统，有潜力弥合差距 哺乳动物内耳和侧线研究之间的联系。这个项目研究毛细胞再生在 斑马鱼内耳在幼虫阶段，在这一点上的感觉器官成为功能齐全， 保持可用于体内成像。我们的目标是识别斑马鱼内耳中的毛细胞前体， 决定它们在再生过程中何时以及如何产生新的毛细胞。在鸟类的内耳中， 在哺乳动物中，支持细胞通过增殖和分化或通过直接转分化再生毛细胞。 在斑马鱼侧线中，仅使用前一种机制。在本提案的目标1中，我们将使用一种新的 毛细胞消融的方法，以确定再生的时程和细胞是否增殖， 转分化或两者都用于斑马鱼内耳。侧线的毛细胞再生 主要由空间上和功能上不同的支持细胞群所控制。是否有类似的支持细胞亚型 存在于斑马鱼的内耳是未知的。在目标2中，我们将验证假定的内耳支持细胞亚型 通过单细胞RNA-seq分析鉴定。然后，我们将产生标记支持细胞的转基因系， 亚型，并使用命运映射来识别毛细胞前体。这些实验将共同描述 斑马鱼毛细胞再生的时间和机制以及支持细胞亚型的多样性 内耳这项工作将有助于建立斑马鱼内耳作为一个优秀的体内模型系统， 支持细胞及其在保守再生机制中的作用的未来深入研究。扩大 我们对动物（如斑马鱼）内耳毛细胞再生机制的了解， 最终导致人类听觉和前庭衰退治疗的进步。