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

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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.