Biosensors for determination of multiple neurotransmitters in vertebrate retina

项目概要：本项目的长期目标是开发一种范式转换的神经传感技术 直接，同时监测多种神经递质的活动，以了解大脑功能。 视网膜被选为我们的模型系统，由于其易于访问和完善的神经生理学 以及迫切需要这种工具来了解神经递质在各种眼科疾病中的作用， 糖尿病视网膜病变视网膜感光细胞（视杆细胞和视锥细胞）将光转化为电信号。的 电信号通过双极细胞传输到视网膜的输出神经元神经节细胞，然后 到大脑。通过这一途径的信号传递是由无长突细胞调节的， 中间神经元有多种类型的无长突细胞，但都能合成和释放神经调质，如 如多巴胺（DA）、γ-氨基丁酸（GABA）和乙酰胆碱（ACh）。特别是多巴胺能 无长突细胞（DAC）共同释放GABA和DA，其在调节视网膜光敏感性中起关键作用 眼睛的发育。星爆无长突细胞共同释放GABA和ACh，启动运动方向 的视觉系统。从历史上看，视网膜神经元和无长突细胞释放神经递质， 间接研究，通过电生理学方法和/或氧化还原检测，使用电分析 使用碳纤维微电极的技术。然而，细胞中的电活动并不总是匹配 神经递质从细胞中释放出来氧化还原方法仅适用于相对少量的分析物 我们构建了一种新型的生物传感器，其采用互补的电化学， 压电传感器，我们的初步结果表明，它可以区分氧化还原和非氧化还原 活跃的神经递质这个R21项目的目标是开发一种小型化的多模态生物传感器， 在真实的时间内以高空间和时间分辨率同时测量多种神经递质，标记- 和无试剂，具有两个目的：1.小型化多模态的设计、制造和表征 电化学（E）和压电传感器（薄膜体声波谐振器（FBAR）（即E-FBAR）） 2：通过监测多巴胺、GABA和ACh验证神经传感探头 在正常和糖尿病视网膜神经元中。本项目的成功完成将证明无试剂， 无标记和实时同时检测氧化还原活性和非氧化还原活性神经递质， 视网膜具有高灵敏度和选择性的多方面信息。这样一个工具将是非常宝贵的研究 旨在了解视网膜神经退行性疾病的原因和机制， 如糖尿病性视网膜病，以及测试用于治疗这些疾病的治疗剂。这本小说 这项技术也可以用来监测大脑中其他重要的神经递质， 了解大脑的功能。我们的团队经验丰富，技术精湛，多学科的专业知识 在电化学和声学生物传感器、微器件和微传感器设计和制造以及视觉 神经科学，以开发和验证拟议的神经传感技术。

Project Summary: The long-term goal of this project is to develop a paradigm-shifting neurosensing technology for direct, simultaneous monitoring of the activity of multiple neurotransmitters for understanding brain function. The retina is selected as our model system due to its easy accessibility and well-established neurophysiology and the urgent needs in such tool to understand the roles of neurotransmitters in various eye diseases such as diabetic retinopathy. Retinal photosensitive cells (rods and cones) convert light into an electrical signal. The electrical signal is transmitted through bipolar cells to ganglion cells, the output neurons of the retina, and then to the brain. Signal transmission through this pathway is modulated by amacrine cells, which are retinal interneurons. There are multiple types of amacrine cells, but all synthesize and release neuromodulators such as dopamine (DA), gamma-aminobutyric acid (GABA) and acetylcholine (ACh). Specifically, dopaminergic amacrine cells (DACs) co-release GABA and DA, which play a critical role in modulating retinal light sensitivity and eye development. Starburst amacrine cells co-release GABA and ACh, which initiates the motion direction of the visual system. Historically, the release of neurotransmitters from retinal neurons and amacrine cells has been studied indirectly, through electrophysiological methods and/or redox detection using electroanalytical techniques employing carbon fiber microelectrodes. However, electrical activity in a cell does not always match the release of neurotransmitter from the cell. Redox methods only work for a relatively small number of analytes such as DA. We have constructed a novel biosensor that employs complementary electrochemical and piezoelectric sensors, and our preliminary results show that it can differentiate between redox and non-redox active neurotransmitters. The objective of this R21 project is to develop a miniaturized multimodal biosensor to measure multiple neurotransmitters simultaneously with high spatial and temporal resolution in real time, label- and reagent-free with two Aims: 1. Design, fabrication, and characterization of a miniaturized multimodal electrochemical (E) and piezoelectric sensor (thin film bulk acoustic resonator (FBAR) (i.e. E-FBAR) neurosensing probe; and 2: Validation of the neurosensing probe through monitoring dopamine, GABA, and ACh in living normal and diabetic retinal neurons. Successful completion of this project will certify a reagent-free, label-free and real-time simultaneously detection of both redox active and non-redox active neurotransmitters in retina with multifaceted information in high sensitivity and selectivity. Such a tool will be invaluable to research aimed at understanding the causes and mechanisms responsible for retinal neurodegenerative diseases such as diabetic retinopathy, and also to test therapeutic agents for the treatment of such diseases. This novel technology could also be adapted to monitor other important neurotransmitters in the brain, increasing our understanding of brain functions. Our well- established, highly skilled, multidisciplinary team has the expertise in electrochemical and acoustic biosensors, microdevice and microsensor design and fabrication, and visual neuroscience to develop and validate the proposed neurosensing technology.