NIDA IRP Technology Resource Initiative Featured Paper

A figure from this article. Image copyright bioRxiv.

Development of a genetically encoded sensor for probing endogenous nociceptin opioid peptide release

Published in bioRxiv.

Authors

Xuehan Zhou, Carrie Stine, Patricia Oliveira Prada, Debora Fusca, Kevin Assoumou, Jan Dernic, Musadiq A Bhat, Ananya S Achanta, Joseph C Johnson, Amanda Loren Pasqualini, Sanjana Jadhav, Corinna A Bauder, Lukas Steuernagel, Luca Ravotto, Dietmar Benke, Bruno Weber, Azra Suko, Richard D Palmiter, Miriam Stoeber, Peter Kloppenburg, Jens C Brüning, Michael R Bruchas, Tommaso Patriarchi

Paper presented by Dr. Agnieszka Sulima and selected by the NIDA TDI Paper of the Month Committee

Publication Brief Description

The nociceptin/orphanin FQ (N/OFQ) opioid peptide and its receptor (NOPR) are critical regulators of motivation, stress, feeding, and sleep, and growing evidence identifies the NOP receptor as a promising target for the treatment of addiction, insomnia, and depression. However, studying the dynamics of the N/OFQ-NOPR peptide system has been challenging due to limitations in existing detection methods.

The research groups led by Michael R. Bruchas and Tommaso Patriarchi developed NOPLight by engineering a human NOPR-based fluorescent sensor with high ligand selectivity, subsecond activation kinetics, and minimal interference with cellular signaling. NOPLight was validated in HEK293T cells, neurons, brain slices, and freely moving mice, demonstrating its ability to detect both exogenous NOPR agonists and endogenous N/OFQ release during behaviors like reward-seeking, aversion, and feeding. This sensor provides a powerful tool for investigating neuropeptide signaling and its role in neural circuits and behavior.

Zhou, Xuehan; Stine, Carrie; Prada, Patricia Oliveira; Fusca, Debora; Assoumou, Kevin; Dernic, Jan; Bhat, Musadiq A; Achanta, Ananya S; Johnson, Joseph C; Pasqualini, Amanda Loren; Jadhav, Sanjana; Bauder, Corinna A; Steuernagel, Lukas; Ravotto, Luca; Benke, Dietmar; Weber, Bruno; Suko, Azra; Palmiter, Richard D; Stoeber, Miriam; Kloppenburg, Peter; Brüning, Jens C; Bruchas, Michael R; Patriarchi, Tommaso

Development of a genetically encoded sensor for probing endogenous nociceptin opioid peptide release Journal Article

In: bioRxiv, 2024, ISSN: 2692-8205.

Abstract | Links

@article{pmid37292957,

title = {Development of a genetically encoded sensor for probing endogenous nociceptin opioid peptide release},

author = {Xuehan Zhou and Carrie Stine and Patricia Oliveira Prada and Debora Fusca and Kevin Assoumou and Jan Dernic and Musadiq A Bhat and Ananya S Achanta and Joseph C Johnson and Amanda Loren Pasqualini and Sanjana Jadhav and Corinna A Bauder and Lukas Steuernagel and Luca Ravotto and Dietmar Benke and Bruno Weber and Azra Suko and Richard D Palmiter and Miriam Stoeber and Peter Kloppenburg and Jens C Brüning and Michael R Bruchas and Tommaso Patriarchi},

url = {https://pubmed.ncbi.nlm.nih.gov/37292957/},

doi = {10.1101/2023.05.26.542102},

issn = {2692-8205},

year  = {2024},

date = {2024-05-01},

urldate = {2024-05-01},

journal = {bioRxiv},

abstract = {Nociceptin/orphanin-FQ (N/OFQ) is a recently appreciated critical opioid peptide with key regulatory functions in several central behavioral processes including motivation, stress, feeding, and sleep. The functional relevance of N/OFQ action in the mammalian brain remains unclear due to a lack of high-resolution approaches to detect this neuropeptide with appropriate spatial and temporal resolution. Here we develop and characterize NOPLight, a genetically encoded sensor that sensitively reports changes in endogenous N/OFQ release. We characterized the affinity, pharmacological profile, spectral properties, kinetics, ligand selectivity, and potential interaction with intracellular signal transducers of NOPLight . Its functionality was established in acute brain slices by exogeneous N/OFQ application and chemogenetic induction of endogenous N/OFQ release from PNOC neurons.  studies with fibre photometry enabled direct recording of NOPLight binding to exogenous N/OFQ receptor ligands, as well as detection of endogenous N/OFQ release within the paranigral ventral tegmental area (pnVTA) during natural behaviors and chemogenetic activation of PNOC neurons. In summary, we show here that NOPLight can be used to detect N/OFQ opioid peptide signal dynamics in tissue and freely behaving animals.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}