Commercialization of Enzyme Modified Carbon-Fiber Electrodes Paired with Voltammetry for Simultaneous Real-Time Monitoring of Electroactive and Non-Electroactive Species at Discrete Brain Locations

摘要 商业上可用的实时分子监测技术旨在选择性地 在给定的记录地点，一次只能测量一个分子。这是一个问题，因为化学物质 大脑中的信号并不是孤立地工作的；相反，神经传递涉及许多化学物质 同时调节神经回路的物种。对这些问题的定量分析 神经化学信号是制定治疗策略的关键第一步 神经/心理障碍，但对具体的神经化学物质如何 彼此相对波动。Sombers实验室已经确定了使用FAST- 扫描循环伏安法(FSCV)和碳纤维微电极的实时检测 多巴胺波动，同时检测非电活性物种，如葡萄糖 和乳酸盐，在相同的记录地点。微生物传感器具有较高的空间和时间分辨率 比目前可用的技术分辨率高，分析物消耗最少，而且它们可以 轻松集成到现有协议中。在第一阶段，开发了一个7微米的探测器，并 商业化，可以同时实时测量多巴胺和葡萄糖， 活体中的单微米级记录位置。软件套件和FSCV教程 发展起来的。在第二阶段，我们将开发和商业化7微米多巴胺/乳酸， 多巴胺/谷氨酸和5-羟色胺/葡萄糖传感器。我们将开发其他教程来 简化对这项重要技术的访问，我们将继续开发软件和 电子产品。总体而言，这个项目是创新的，因为它通过利用 酶促产生的过氧化氢固有的氧化还原活性以识别靶向非电活性 物种，即使在存在电活性分子的情况下，通常被排除为 干扰物。它意义重大，因为它结合了两个最先进的和很好的特点 智能、直截了当、史无前例的神经化学监测技术 举止。最终，这个项目将为社区提供一个可以使用的传感器套件 研究一系列关键分析物在复杂的生理过程中的相互作用 从基本的内分泌功能到动力。它有望对…产生变革性的影响 神经科学让对大脑功能的不同方面感兴趣的研究人员更好地 了解这些特定的神经化学物质如何在不同的大脑位置共同波动。

ABSTRACT Commercially available, real-time molecular monitoring technologies are designed to selectively measure only one molecule at a time at a given recording site. This is a problem because chemical signals in the brain do not work in isolation; rather, neurotransmission involves many chemical species simultaneously working to modulate neural circuits. Quantitative analysis of these neurochemical signals is a critical first step when developing therapeutic strategies to treat neurological/psychological disorders, but little is known about how specific neurochemicals fluctuate relative to one another. The Sombers Lab has established the feasibility of using fast- scan cyclic voltammetry (FSCV) and carbon-fiber microelectrodes for the real-time detection of dopamine fluctuations while simultaneously detecting non-electroactive species, such as glucose and lactate, at the same recording site. The microbiosensors have higher spatial and temporal resolution than currently available technologies, minimal analyte consumption, and they can be easily integrated into existing protocols. During Phase I, a 7-um probe was developed and commercialized that can simultaneously measure dopamine and glucose in real time, at single micron-scale recording sites in vivo. A software suite and FSCV tutorials were developed. In Phase II, we will develop and commercialize 7-um dopamine/lactate, dopamine/glutamate, and serotonin/glucose sensors. We will develop additional tutorials to simplify access to this important technology, and we will continue to develop software and electronics. Overall, this project is innovative, because it departs from the status quo by utilizing the redox activity inherent to enzymatically generated H2O2 to identify targeted non-electroactive species, even in the presence of electroactive molecules that are typically excluded as interferents. It is significant, because it combines two state-of-the-art and well-characterized technologies for neurochemical monitoring in a clever, straightforward, and unprecedented manner. Ultimately, this project will provide the community with a sensor suite that can be used to study the interplay of a range of critical analytes in complex physiological processes ranging from basic endocrine function to motivation. It promises to have a transformative effect on neuroscience by allowing researchers interested in diverse aspects of brain function to better understand how these specific neurochemicals co-fluctuate in discrete brain locations.