Neuron-Glia Interactions in the Cochlea

耳蜗中神经元-神经胶质细胞的相互作用

• 批准号: 10611512
• 负责人: Lisa Goodrich
• 金额: $ 52.46万
• 依托单位: HARVARD MEDICAL SCHOOL
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-05-01 至 2027-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10611512
• 关键词: Ablation; Acute; Affect; Age; Auditory; Axon; Behavior; Brain; Cells; Central Nervous System; Cochlea; Connective Tissue; Cues; Cytoskeleton; Data; Detection; Development; Diphtheria Toxin; Disease; Embryo; Embryonic Development; Endowment; Event; Family; Freezing; GATA2 transcription factor; GATA3 gene; Gene Expression; Genes; Genetic; Glial Differentiation; Glioblastoma; Goals; Hair Cells; Hearing; Human; Image; In Situ Hybridization; Individual; Ions; Knockout Mice; Knowledge; Label; Labyrinth; Learning; Mediating; Molecular; Molecular Genetics; Morphology; Mouse Strains; Movement; Mus; Mutant Strains Mice; Mutate; Mutation; Natural regeneration; Neural Crest; Neural Crest Cell; Neurites; Neuroglia; Neurons; Noise; Organ of Corti; Pattern; Peripheral; Peripheral Nervous System; Phenotype; Play; Population; Process; Prognosis; Property; Publishing; Radial; Reporting; Role; Schwann Cells; Sensorineural Hearing Loss; Shapes; Signal Transduction; Source; Supporting Cell; Syndrome; System; Testing; Toxin; Viral; Visualization; cell type; cochlear development; conditional knockout; experience; experimental study; follow-up; genetic approach; glial cell development; hearing impairment; hereditary hearing loss; in vivo; insight; migration; mutant; myelination; nerve supply; neuron development; neuron loss; neuronal cell body; neuronal survival; otoconia; permissiveness; prevent; progenitor; programs; progressive hearing loss; repaired; single-cell RNA sequencing; somatosensory; sound; spiral ganglion; stem-like cell; time use; tissue fixing; transcription factor; transmission process; Ablation; Acute; Affect; Age; Auditory; Axon; Behavior; Brain; Cells; Central Nervous System; Cochlea; Connective Tissue; Cues; Cytoskeleton; Data; Detection; Development; Diphtheria Toxin; Disease; Embryo; Embryonic Development; Endowment; Event; Family; Freezing; GATA2 transcription factor; GATA3 gene; Gene Expression; Genes; Genetic; Glial Differentiation; Glioblastoma; Goals; Hair Cells; Hearing; Human; Image; In Situ Hybridization; Individual; Ions; Knockout Mice; Knowledge; Label; Labyrinth; Learning; Mediating; Molecular; Molecular Genetics; Morphology; Mouse Strains; Movement; Mus; Mutant Strains Mice; Mutate; Mutation; Natural regeneration; Neural Crest; Neural Crest Cell; Neurites; Neuroglia; Neurons; Noise; Organ of Corti; Pattern; Peripheral; Peripheral Nervous System; Phenotype; Play; Population; Process; Prognosis; Property; Publishing; Radial; Reporting; Role; Schwann Cells; Sensorineural Hearing Loss; Shapes; Signal Transduction; Source; Supporting Cell; Syndrome; System; Testing; Toxin; Viral; Visualization; cell type; cochlear development; conditional knockout; experience; experimental study; follow-up; genetic approach; glial cell development; hearing impairment; hereditary hearing loss; in vivo; insight; migration; mutant; myelination; nerve supply; neuron development; neuron loss; neuronal cell body; neuronal survival; otoconia; permissiveness; prevent; progenitor; programs; progressive hearing loss; repaired; single-cell RNA sequencing; somatosensory; sound; spiral ganglion; stem-like cell; time use; tissue fixing; transcription factor; transmission process

Project Summary In this project, we will investigate the cellular and molecular events that allow cochlear glia to shape and sustain auditory circuits for a lifetime of hearing. Like the other glia of the peripheral nervous system, cochlear glia develop from the neural crest, a highly migratory and plastic population of progenitors that produce neurons and connective tissue as well as satellite glia and Schwann cells. Despite their common origin with other peripheral glia, the glia that ultimately populate the cochlea take on some unique properties that are essential for the sense of hearing, such as the ability to myelinate spiral ganglion neuron (SGNs) cell bodies. In addition, cochlear glia provide crucial trophic support for the highly active SGNs and also clear away excess ions and transmitter that could otherwise be damaging. Finally, whereas glial precursors generally migrate along axons, in the cochlea, the glial precursors migrate ahead of the SGN neurites and appear to facilitate efficient formation of orderly radial bundles along the tonotopic axis. Our overall goal is to learn how cochlear glia acquire these properties. Based on data from our lab and others, we hypothesize that the transcription factors Gata2 and Gata3 act separately in developing glia and SGNs to coordinate interactions needed for cochlear wiring and function. In support of this idea, we found that Gata2 is expressed by cochlear glia but not by glia of the somatosensory system. Additionally, previous analysis of Gata2 mutant mice revealed hearing deficits and SGN loss that could be due to unrecognized effects on the glia. On the other hand, we observed that Gata3 mutant SGNs extend their neurites as if they are unable to interact with the glia, resulting in disorderly wiring that mimics what occurs when glia are depleted from the developing cochlea. We will follow up on these observations by using molecular genetic approaches in the mouse to characterize glial organization and its effects on SGN neurite outgrowth (Aim 1), to determine the role of Gata2 in cochlear glia development (Aim 2), and to compare GATA-dependent programs of gene expression in SGNs and glia (Aim 3). For Aim 1, we will use genetic and viral approaches to visualize glia and disrupt their ability to interact with developing SGN neurites, as assessed in fixed tissue and by time-lapse imaging. For Aim 2, we will delete Gata2 from cochlear glia and assess effects on cochlear wiring, on glial differentiation and function, on SGN survival, and on auditory function, as assessed by recording ABRs and DPOAEs. For Aim 3, we will perform scRNA-sequencing of embryonic Gata2 and Gata3 mutant cochleas, both to identify downstream genes that may mediate neuron-glia interactions and to learn how neurons and glia are affected by each other during development. Together, these studies will advance our knowledge of cochlear glia development and function, with direct implications for hearing loss, including that associated with Gata2 (Emberger Syndrome) and Gata3 (HDR Syndrome) mutations in humans.

项目摘要 在这个项目中，我们将研究允许人工耳蜗塑造和维持的细胞和分子事件 听觉电路一生的听力。就像外周神经系统的其他神经胶质一样 从神经rest发展，神经rest是产生神经元和 结缔组织以及卫星胶质细胞和雪旺细胞。尽管它们与其他外围有共同的起源 神经胶质，最终填充耳蜗具有一些独特特性的神经胶质 听力，例如能够髓鞘的螺旋神经神经元（SGNS）细胞体的能力。另外，人工耳蜗 为高度活性的SGN提供至关重要的营养支持，并清除多余的离子和发射器 否则可能会造成破坏。最后，而神经胶质前体通常沿着轴突迁移，而在耳蜗中，但 神经胶质前体在SGN神经突迁移，似乎有助于有序地形成有序的径向 沿吨位轴捆绑。我们的总体目标是了解人工耳蜗如何获得这些特性。基于 关于我们实验室和其他人的数据，我们假设转录因子GATA2和GATA3分别起作用 在开发神经胶质和SGN以协调耳蜗接线和功能所需的相互作用。支持 这个想法，我们发现GATA2是由人工耳蜗表达的，而不是通过体感系统的神经胶质表示。 此外，对GATA2突变小鼠的先前分析表明，听力缺陷和SGN损失可能应得 无法识别对神经胶质的影响。另一方面，我们观察到GATA3突变体SGN延伸 神经突变好像无法与神经胶质相互作用，从而导致无序的接线，以模仿发生 神经胶质是从发育中的耳蜗中耗尽的。我们将通过使用分子来跟进这些观察结果 小鼠中的遗传方法表征神经胶质组织及其对SGN神经突生长的影响 （AIM 1），确定GATA2在人工耳蜗发育中的作用（AIM 2），并比较GATA依赖性 SGNS和GLIA中的基因表达程序（AIM 3）。对于AIM 1，我们将使用遗传和病毒方法来 可视化神经胶质并破坏其与发育中的SGN神经突相互作用的能力，如固定组织和 通过延时成像。对于AIM 2，我们将从人工耳蜗中删除GATA2，并评估对耳蜗接线的影响， 关于通过记录ABR评估的胶质分化和功能，SGN存活和听觉功能 和dpoaes。对于AIM 3，我们将执行胚胎GATA2和GATA3突变体耳蜗的SCRNA测序， 两者都可以识别可能介导神经元相互作用的下游基因，并了解神经元和神经胶质 在开发过程中彼此影响。这些研究将共同​​提高我们对人工耳蜗的了解 GliA的开发和功能，对听力损失有直接影响，包括与GATA2相关的听力损失 （Emberger综合征）和GATA3（HDR综合征）突变。