Identifying new sensors for in vivo cochlear imaging

摘要：外周听觉系统通过耳蜗深埋在颞骨内。 被骨头包围，受压，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。我们已经开发出一种 手术入路可以接触到耳蜗骨，我们已经开发出一种化学机械 在不损失听力功能的情况下用耳蜗造像窗的方法。我们有 发展到需要调查现有的和新的基因编码传感器的程度 以监测细胞功能。我们与基因编码传感器方面的专家进行了合作。 Michael Lin将开发使用ASAP4靶向毛细胞和螺旋神经节神经元的策略 电压传感器。这项提议的目标是描述螺旋神经节的感受器特性。 在体外和活体神经元中确定最佳传感器和最佳靶向传感器的手段 胞体和突触。我们进一步表征了钙传感器，如GCaMP 6s和f在这两个 内毛细胞和外毛细胞，因为它们以cre激活的形式存在，可以选择性地在 用于活体成像的毛细胞。我们还将在体外鉴定毛细胞中的电压传感器，并 如果这些传感器被证明比GCaMP更好，就转移到体内。这一准备工作以及 传感器特性将开启耳蜗内多细胞功能成像的新纪元。

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.