Development and Validation of an NPY-sensitive Microelectrode for Measuring NPY Release from Hippocampus

用于测量海马 NPY 释放的 NPY 敏感微电极的开发和验证

• 批准号: 10391927
• 负责人: Lisandro Federico Cunci Perez
• 金额: $ 20.33万
• 依托单位: UNIVERSITY COLLEGE OF TURABO (CAGUAS,PR)
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-21 至 2023-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10391927
• 关键词: Acute; Adsorption; Advanced Development; Affect; Alcoholism; Anti-Anxiety Agents; Anxiety; Anxiety Disorders; Behavior; Binding; Biological Process; Brain; Brain Chemistry; Catecholamines; Cells; Chemicals; Circadian Rhythms; Communities; Detection; Development; Disease; Electric Stimulation; Electrodes; Electrophysiology (science); Environment; Enzyme-Linked Immunosorbent Assay; Feeding behaviors; Frequencies; Future; Goals; Hippocampus (Brain); Joints; Kinetics; Knockout Mice; Knowledge; Learning; Measurement; Measures; Memory; Methods; Microelectrodes; Modification; Monitor; Mus; Neurobiology; Neuropeptides; Neurotransmitters; Obesity; Oxidases; Pain; Pathway interactions; Periodicity; Pharmacology; Physiological; Platinum; Post-Traumatic Stress Disorders; Process; Property; Reaction; Regulation; Research; Research Personnel; Research Proposals; Resolution; Role; Sampling; Scanning; Signal Transduction; Slice; Specificity; Spectrum Analysis; Structure; Surface; Surface Properties; Synaptic Transmission; Techniques; Technology; Testing; Tetracyclines; Time; Tissues; Transgenic Mice; Validation; Wild Type Mouse; addiction; aptamer; base; brain tissue; carbon fiber; chemical property; depressive symptoms; detection limit; electric impedance; experimental study; human disease; improved; in vivo; knock-down; mood regulation; nervous system disorder; neuropeptide Y; neurotransmitter release; novel; novel strategies; optogenetics; overexpression; physical property; response; stress disorder; synaptic function; temporal measurement; tool

This research proposal aims to develop and validate microelectrode that is sensitive to neuropeptide Y (NPY) for measuring the release of NPY from hippocampus. In this way, we will be able to find correlations between NPY levels with anxiety disorders. In order to do this, two electrochemical strategies have been devised to monitor biomolecules in real-time. Non-electroactive molecules in the brain are difficult to measure with high temporal and spatial resolution and neuropeptides have been a challenge. Electrochemical-based techniques are powerful and can be used to measure the physical and chemical properties of the surface and they have been vastly used for the detection of molecules with very low detection limits. The combination of highly selective aptamers with fast scan cyclic voltammetry and continuous electrochemical impedance measurements will provide two novel strategies to understand the presence of NPY in the CA1 region. Different molecules that are potentially released together with NPY will be measured using the developed microelectrodes to prove selectivity. Genetically modified mice will be under and overregulated using tetracyclines to change NPY levels and confirm the measurement using the developed NPY-sensitive microelectrodes. Platinum microelectrodes measuring up to 25 micrometers will provide the appropriate substrate for the adsorption and desorption of molecules as well as the aptamer modification to filter NPY signals from other confounding molecules. Concomitant electrochemical and electrophysiological measurement in CA1 will be done to filter NPY from the different other signals measured. The confirmation of the effects of NPY will be tested recording fEPSPs in response to low-frequency electrical stimulation in the SC and TA pathway. In order to validate NPY levels, ELISA will be used to compare the measurements done with our developed NPY-sensitive microelectrodes in hippocampal extracts. Electrochemical impedance spectroscopy and fast scan cyclic voltammetry have shown to be important techniques that allow the measurement of faradaic as well as non-faradaic currents providing a picture of the electroactive species as well as non-electroactive species that interact with the electrode’s surfaces. The combination of fast scan cyclic voltammetry with electrochemical impedance measurements will empower the research community using microelectrodes for real-time measurement of biomolecules such as neurotransmitters and neuropeptides.

本研究旨在研制和验证对神经肽Y敏感的微电极 (NPY)用于测量海马体中神经肽Y的释放。通过这种方式，我们将能够找到相关性 神经肽Y水平与焦虑症之间的关系为了做到这一点，两种电化学策略已经被研究。 用来实时监测生物分子大脑中的非电活性分子很难测量 具有高的时间和空间分辨率和神经肽一直是一个挑战。基于电化 技术是强大的，可用于测量表面的物理和化学性质， 它们已广泛用于检测具有非常低检测限的分子。的组合 具有快速扫描循环伏安法和连续电化学阻抗的高选择性适体 测量将提供两种新的策略，以了解在CA 1区的存在NPY。 不同的分子，可能与神经肽Y一起释放，将使用开发的测量 微电极来证明选择性。转基因小鼠将被过度调节， 四环素改变NPY水平，并使用开发的NPY敏感的 微电极测量到25微米的铂微电极将提供适当的 用于吸附和解吸分子以及适配体修饰以过滤NPY的基底 来自其他干扰分子的信号伴随电化学和电生理学 将进行CA 1中的测量，以从测量的不同其他信号中过滤NPY。的确认 将测试NPY的作用，记录响应SC中低频电刺激的fEPSP 和TA通路。为了验证NPY水平，将使用ELISA来比较用以下方法进行的测量： 我们开发的海马提取物中的NPY敏感微电极。 电化学阻抗谱和快速扫描循环伏安法已被证明是重要的 允许测量法拉第电流以及非法拉第电流的技术提供了 电活性物质以及与电极表面相互作用的非电活性物质。的 快速扫描循环伏安法与电化学阻抗测量的组合将使 使用微电极实时测量生物分子的研究团体， 神经递质和神经肽。