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Papers of the Week


Papers: 11 Feb 2023 - 17 Feb 2023

RESEARCH TYPE:
Basic Science, Methodology


Animal Studies, Molecular/Cellular, Pharmacology/Drug Development


2023 Feb 03


bioRxiv


36778492

An electrochemical approach for rapid, sensitive, and selective detection of dynorphin.

Authors

Conway SM, Kuo CC, Gardiner W, Wu RN, Thang LV, Gereau GB, Cirrito JR, Yuede CM, McCall JG, Al-Hasani R

Abstract

The endogenous opioid peptide systems are critical for analgesia, reward processing, and affect, but research on their release dynamics and function has been challenging. Here, we have developed microimmunoelectrodes (MIEs) for the electrochemical detection of opioid peptides using square-wave voltammetry. Briefly, a voltage is applied to the electrode to cause oxidation of the tyrosine residue on the opioid peptide of interest, which is detected as current. To provide selectivity to these voltammetric measurements, the carbon fiber surface of the MIE is coated with an antiserum selective to the opioid peptide of interest. To test the sensitivity of the MIEs, electrodes are immersed in solutions containing different concentrations of opioid peptides, and peak oxidative current is measured. We show that dynorphin antiserum-coated electrodes are sensitive to increasing concentrations of dynorphin in the attomolar range. To confirm selectivity, we also measured the oxidative current from exposure to tyrosine and other opioid peptides in solution. Our data show that dynorphin antiserum-coated MIEs are sensitive and selective for dynorphin with little to no oxidative current observed in met-enkephalin and tyrosine solutions. Additionally, we demonstrate the utility of these MIEs in an brain slice preparation using bath application of dynorphin as well as optogenetic activation of dynorphin release. Future work aims to use MIEs for real-time, rapid detection of endogenous opioid peptide release in awake, behaving animals.