Identifying new sensors for in vivo cochlear imaging

翻译后摘要：周围听觉系统是深埋在颞骨与耳蜗 被骨头包围着，压力很大，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 手术方法，允许进入耳蜗，我们已经开发了化学机械 该方法可以在不丧失听力功能的情况下用耳蜗创建成像窗口。我们有 发展到我们需要调查现有的和新的基因编码传感器的地步， 来监控细胞功能我们与一位基因编码传感器专家合作， Michael Lin开发ASAP4靶向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 神经元在体外和体内，以确定最佳的传感器和最佳的手段，目标传感器， 索马和突触。我们进一步表征了钙传感器，如GCaMP 6s和f， 内毛细胞和外毛细胞以CRE活化形式存在， 用于体内成像的毛细胞。我们还将表征体外毛细胞中的电压传感器， 如果这些传感器证明比GCaMP更好，则转移到体内。这种准备沿着 传感器表征将开启耳蜗内多细胞功能成像的新时代。

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.