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

基本信息

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
项目类别：
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

项目摘要

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.

该研究建议旨在开发和验证对神经肽y敏感的微电极（NPY）用于测量NPY从海马释放。这样，我们将能够找到相关性在NPY水平之间患有焦虑症。为此，已经有两种电化学策略设计用于实时监测生物分子。大脑中的非电动活性分子很难测量由于临时和空间分辨率高，神经肽一直是一个挑战。基于电化学技术功能强大，可用于测量表面的物理和化学特性它们已被广泛用于检测具有非常低检测极限的分子。结合具有快速扫描循环伏安法和连续电化学阻抗的高度选择性适体测量将提供两种新型策略，以了解NPY在CA1地区的存在。可能与NPY一起释放的不同分子将使用开发的微电极证明选择性。转基因的小鼠将在使用并过度调节四环素以改变NPY水平并使用开发的NPY敏感性确认测量微电极。铂微电极测量多达25微米将提供适当的底物增加分子的吸附和解吸以及APATMER修饰以过滤NPY 来自其他混杂分子的信号。伴随的电化学和电生理学将在CA1中进行测量，以从测量的其他其他信号中过滤NPY。确认将测试NPY的效果，以记录FEPSP，以响应SC中的低频电模拟。和TA路径。为了验证NPY水平，ELISA将用于比较完成的测量我们在海马提取物中开发的NPY敏感微电极。电化学阻抗光谱和快速扫描循环伏安法已证明很重要允许测量法拉达语以及非瓦拉达型电流的技术，可提供与电极表面相互作用的电活性物种以及非电活性物种。快速扫描循环伏安法与电化学阻抗测量的结合将增强研究社区使用微电极进行实时测量生物分子（例如神经递质和神经肽。