Neural Probe for high spatial and temporal resolution detection of cocaine and su

描述（由申请人提供）：本提案的具体目的是测试开发一种持久的、可植入的探针的可行性，用于快速测量大脑中的多种神经化学物质。目前，神经科学研究仅限于三种测量体内神经化学物质浓度的技术；微透析可以在相对较长的时间内（5-20分钟）获得平均浓度，基于酶的生物传感器可以在相对较大的空间区域内每秒检测一种神经化学物质（500 μ m长度的电极），碳纤维微电极可以通过快速扫描循环伏安法（FSCV）检测多巴胺。需要一种新的工具来快速检测细胞水平上具有空间分辨率的多种神经化学物质的浓度。这样的工具将允许神经科学研究人员对疾病状态和行为背后的机制提出新的问题，比如药物消耗。提出的神经探针通过每4秒以50米的空间分辨率检测两种神经化学物质来满足这一需求。提出的探针将检测可卡因和物质P，一种隐含在可卡因成瘾中的神经肽（Kampman, 2010）。2008年，有报告指出，140万美国人符合滥用或依赖可卡因的标准，这与暴力和监禁有关（NIDA 2010; Nyamathi et al., 2012）。SB微系统公司已经开发出一种专有的MEMS工艺，用于制造可植入的多位点神经探针，用于研究大鼠脑。我们现有的探针具有比现有酶基电极提高10倍空间分辨率所需的特征尺寸。提出的探针将建立在我们现有的平台上，通过分子功能化探针位点表面，用于检测特定的神经化学物质。多种神经化学物质的检测将通过不同的神经化学特异性检测分子到相邻的探针位点来实现。我们的第一阶段提案将通过以下方式确定这些探针商业化的可行性；1)通过调整适配体分子修饰、固定化技术和电信号检测来改进功能化探针的制造，以达到最佳的灵敏度和时间响应；2)开发基于检测的恒电位器电路。接下来，我们将3)功能化探针以检测可卡因和P物质适配体的多种分析物，4)将探针植入大鼠体内进行体内数据收集。第一阶段可行性研究的成功与否将取决于该探针能否准确检测出生理上相关的可卡因和P物质浓度，这些探针在体内稳定2天。在第二阶段，我们计划开发更多的适体，可以应用于我们的探针检测2种以上的神经递质。我们将使用稳健设计的原则，把我们的原型变成商业产品。所附的支持信表明，我们可能能够将第一阶段的成功原型出售给神经科学研究人员。

DESCRIPTION (provided by applicant): The specific aim of this proposal is to test the feasibility of developing a long-lasting, implantable probe for rapid measurement of multiple neurochemicals in the brain. Currently, neuroscience research is limited to three techniques for measuring the concentrations of neurochemicals in vivo; microdialysis to obtain average concentrations over a relatively long time period (5-20 minutes), enzyme-based biosensors to detect a single neurochemical every second over a relatively large spatial area (500 ¿m length electrode), and carbon-fiber microelectrodes to detect dopamine with fast scan cyclic voltammetry (FSCV). A new tool is required for rapid detection of concentrations of multiple neurochemicals with spatial resolution on the cellular level. Such a tool would allow neuroscience researchers to ask new questions about the mechanisms behind disease states and behaviors, such as drug consumption. The proposed neural probe fulfills this need by detecting two neurochemicals every 4 seconds with 50 ¿m spatial resolution. The proposed probe will detect cocaine and substance P, a neuropeptide implied in cocaine addiction (Kampman, 2010). In 2008, reports stated that 1.4 million Americans meet the criteria for abuse or dependence on cocaine, which is associated with violence and incarceration (NIDA 2010; Nyamathi et al., 2012). SB Microsystems has already developed a proprietary MEMS process for fabricating implantable, multi-site neural probes for studying the rat brain. Our existing probes have the feature size necessary for a 10-fold spatial resolution improvement over the available enzyme-based electrodes. The proposed probe will build on our existing platform by functionalizing the probe site surfaces with molecules for the detection of specific neurochemicals. Detection of multiple neurochemicals will be achieved by patterning different neurochemical- specific detection molecules onto adjacent probe sites. Our Phase I proposal will determine feasibility for commercialization of these probes by; 1) improving functionalized probe fabrication by adjusting aptamer molecule modifications, immobilization technique, and electrical signal detection to achieve the best possible sensitivity and time response and 2) developing a Potentiostat circuit based for detection. Next, we will 3) functionalize the probe to detect multiple analytes with the cocaine and substance P aptamers and 4) implant probes into rats for in vivo data collection. Success in this Phase I feasibility study will be determined by te accurate detection of physiologically relevant concentrations of cocaine and substance P by probes that are stable in vivo for 2 days. In Phase II, we plan to develop more aptamers that can be applied to our probes for the detection of more than 2 neurotransmitters. We will use principles of robust design to turn our prototype into a commercial product. The attached letters of support indicate that we may be able to sell a successful prototype from Phase I to neuroscience researchers.