Identifying new sensors for in vivo cochlear imaging

识别用于体内耳蜗成像的新传感器

• 批准号: 10433182
• 负责人: Anthony J Ricci
• 金额: $ 23.94万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-05-02 至 2024-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10433182
• 关键词: Adult; Apical; Auditory; Auditory system; Biological; Calcium; Calibration; Cell physiology; Cells; Chemicals; Cochlea; DNA Sequence Alteration; Environment; Family; Fiber; Functional Imaging; Generations; Goals; Hair Cells; Image; In Vitro; Individual; Inner Hair Cells; Interferometry; Investigation; Kinetics; Label; Labyrinth; Lasers; Maps; Measurement; Mechanical Stimulation; Mechanics; Methods; Molecular; Molecular Biology; Molecular Genetics; Monitor; Mus; Neurons; Operating System; Operative Surgical Procedures; Optical Coherence Tomography; Optics; Outer Hair Cells; Output; Pattern; Peripheral; Physiologic Monitoring; Preparation; Presynaptic Terminals; Property; Resolution; Resources; Signal Transduction; Specificity; Surveys; Synapses; Synapsins; Synaptic Potentials; System; Technology; Temporal bone structure; Time; Variant; Viral; Work; analytical tool; base; bone; cell type; cellular imaging; deafness; experimental study; ganglion cell; hearing impairment; in vivo; in vivo imaging; in vivo monitoring; neuronal cell body; novel; pressure; presynaptic; promoter; response; sensor; sound; spiral ganglion; technology development; tool; two-photon; vibration; voltage; voltage clamp

Abstract: The peripheral auditory system is buried deep within the temporal bone with a cochlea surrounded by bone, pressurized and exquisitely sensitive to mechanical stimulation. These properties have made in vivo imaging of multicellular function difficult. We have developed a surgical approach that allows access to the cochlea and we have developed a chemo-mechanical approach to create imaging windows with the cochlea without loss of hearing function. We have progressed to the point where we need to survey existing and novel genetically encoded sensors to monitor cell function. We have partnered with an expert in genetically encoded sensors, Dr. Michael Lin to develop strategies to target hair cells and spiral ganglia neurons with ASAP4 voltage sensors. The goal of this proposal is to characterize sensor properties in spiral ganglia neurons in vitro and in vivo to identify the best sensor and the best means to target the sensor to soma and synapses. We further are characterizing calcium sensors like GCaMP 6s and f in both inner and outer hair cells as these exist in cre activated forms that can be selectively activated in hair cells for in vivo imaging. We will also characterize voltage sensors in hair cells in vitro and move to in vivo if these sensors prove better than the GCaMPs. This preparation along with the sensor characterization will open a new era in multicellular functional imaging within the cochlea.

摘要： 外周听觉系统深埋在颞骨内，有耳蜗。 被骨头包围，加压并且对机械刺激非常敏感。这些 这些特性使得多细胞功能的体内成像变得困难。我们开发了一个 允许进入耳蜗的手术方法，我们开发了一种化学机械方法 一种利用耳蜗创建成像窗口而不丧失听力功能的方法。我们有 进展到我们需要调查现有和新型基因编码传感器的地步 监测细胞功能。我们与基因编码传感器专家 Dr. 合作。 Michael Lin 将利用 ASAP4 开发针对毛细胞和螺旋神经节神经元的策略 电压传感器。该提案的目标是表征螺旋神经节中的传感器特性 体外和体内神经元以确定最佳传感器以及将传感器定位到的最佳方法 体细胞和突触。我们进一步对 GCaMP 6s 和 f 等钙传感器进行了表征 内毛细胞和外毛细胞以 cre 激活形式存在，可以选择性地激活 用于体内成像的毛细胞。我们还将在体外表征毛细胞中的电压传感器， 如果这些传感器被证明比 GCaMP 更好，则转向体内。此准备工作连同 传感器表征将开启耳蜗内多细胞功能成像的新时代。