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Modulation of sleep behavior in zebrafish larvae by pharmacological targeting of the orexin receptor

New pharmacological approaches that target orexin receptors (OXRs) are being developed to treat sleep disorders such as insomnia and narcolepsy, with fewer side effects than existing treatments. Orexins are neuropeptides that exert excitatory effects on postsynaptic neurons via the OXRs, and are imp...

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Autores principales: Pardon, Marie, Claes, Pieter, Druwé, Sarah, Martini, Murielle, Siekierska, Aleksandra, Menet, Christel, de Witte, Peter A. M., Copmans, Daniëlle
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9632972/
https://www.ncbi.nlm.nih.gov/pubmed/36339591
http://dx.doi.org/10.3389/fphar.2022.1012622
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author Pardon, Marie
Claes, Pieter
Druwé, Sarah
Martini, Murielle
Siekierska, Aleksandra
Menet, Christel
de Witte, Peter A. M.
Copmans, Daniëlle
author_facet Pardon, Marie
Claes, Pieter
Druwé, Sarah
Martini, Murielle
Siekierska, Aleksandra
Menet, Christel
de Witte, Peter A. M.
Copmans, Daniëlle
author_sort Pardon, Marie
collection PubMed
description New pharmacological approaches that target orexin receptors (OXRs) are being developed to treat sleep disorders such as insomnia and narcolepsy, with fewer side effects than existing treatments. Orexins are neuropeptides that exert excitatory effects on postsynaptic neurons via the OXRs, and are important in regulating sleep/wake states. To date, there are three FDA-approved dual orexin receptor antagonists for the treatment of insomnia, and several small molecule oral OX2R (OXR type 2) agonists are in the pipeline for addressing the orexin deficiency in narcolepsy. To find new hypnotics and psychostimulants, rodents have been the model of choice, but they are costly and have substantially different sleep patterns to humans. As an alternative model, zebrafish larvae that like humans are diurnal and show peak daytime activity and rest at night offer several potential advantages including the ability for high throughput screening. To pharmacologically validate the use of a zebrafish model in the discovery of new compounds, we aimed in this study to evaluate the functionality of a set of known small molecule OX2R agonists and antagonists on human and zebrafish OXRs and to probe their effects on the behavior of zebrafish larvae. To this end, we developed an in vitro IP-One Homogeneous Time Resolved Fluorescence (HTRF) immunoassay, and in vivo locomotor assays that record the locomotor activity of zebrafish larvae under physiological light conditions as well as under dark-light triggers. We demonstrate that the functional IP-One test is a good predictor of biological activity in vivo. Moreover, the behavioral data show that a high-throughput assay that records the locomotor activity of zebrafish throughout the evening, night and morning is able to distinguish between OXR agonists and antagonists active on the zebrafish OXR. Conversely, a locomotor assay with alternating 30 min dark-light transitions throughout the day is not able to distinguish between the two sets of compounds, indicating the importance of circadian rhythm to their pharmacological activity. Overall, the results show that a functional IP-one test in combination with a behavioral assay using zebrafish is well-suited as a discovery platform to find novel compounds that target OXRs for the treatment of sleep disorders.
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spelling pubmed-96329722022-11-04 Modulation of sleep behavior in zebrafish larvae by pharmacological targeting of the orexin receptor Pardon, Marie Claes, Pieter Druwé, Sarah Martini, Murielle Siekierska, Aleksandra Menet, Christel de Witte, Peter A. M. Copmans, Daniëlle Front Pharmacol Pharmacology New pharmacological approaches that target orexin receptors (OXRs) are being developed to treat sleep disorders such as insomnia and narcolepsy, with fewer side effects than existing treatments. Orexins are neuropeptides that exert excitatory effects on postsynaptic neurons via the OXRs, and are important in regulating sleep/wake states. To date, there are three FDA-approved dual orexin receptor antagonists for the treatment of insomnia, and several small molecule oral OX2R (OXR type 2) agonists are in the pipeline for addressing the orexin deficiency in narcolepsy. To find new hypnotics and psychostimulants, rodents have been the model of choice, but they are costly and have substantially different sleep patterns to humans. As an alternative model, zebrafish larvae that like humans are diurnal and show peak daytime activity and rest at night offer several potential advantages including the ability for high throughput screening. To pharmacologically validate the use of a zebrafish model in the discovery of new compounds, we aimed in this study to evaluate the functionality of a set of known small molecule OX2R agonists and antagonists on human and zebrafish OXRs and to probe their effects on the behavior of zebrafish larvae. To this end, we developed an in vitro IP-One Homogeneous Time Resolved Fluorescence (HTRF) immunoassay, and in vivo locomotor assays that record the locomotor activity of zebrafish larvae under physiological light conditions as well as under dark-light triggers. We demonstrate that the functional IP-One test is a good predictor of biological activity in vivo. Moreover, the behavioral data show that a high-throughput assay that records the locomotor activity of zebrafish throughout the evening, night and morning is able to distinguish between OXR agonists and antagonists active on the zebrafish OXR. Conversely, a locomotor assay with alternating 30 min dark-light transitions throughout the day is not able to distinguish between the two sets of compounds, indicating the importance of circadian rhythm to their pharmacological activity. Overall, the results show that a functional IP-one test in combination with a behavioral assay using zebrafish is well-suited as a discovery platform to find novel compounds that target OXRs for the treatment of sleep disorders. Frontiers Media S.A. 2022-10-10 /pmc/articles/PMC9632972/ /pubmed/36339591 http://dx.doi.org/10.3389/fphar.2022.1012622 Text en Copyright © 2022 Pardon, Claes, Druwé, Martini, Siekierska, Menet, de Witte and Copmans. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Pharmacology
Pardon, Marie
Claes, Pieter
Druwé, Sarah
Martini, Murielle
Siekierska, Aleksandra
Menet, Christel
de Witte, Peter A. M.
Copmans, Daniëlle
Modulation of sleep behavior in zebrafish larvae by pharmacological targeting of the orexin receptor
title Modulation of sleep behavior in zebrafish larvae by pharmacological targeting of the orexin receptor
title_full Modulation of sleep behavior in zebrafish larvae by pharmacological targeting of the orexin receptor
title_fullStr Modulation of sleep behavior in zebrafish larvae by pharmacological targeting of the orexin receptor
title_full_unstemmed Modulation of sleep behavior in zebrafish larvae by pharmacological targeting of the orexin receptor
title_short Modulation of sleep behavior in zebrafish larvae by pharmacological targeting of the orexin receptor
title_sort modulation of sleep behavior in zebrafish larvae by pharmacological targeting of the orexin receptor
topic Pharmacology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9632972/
https://www.ncbi.nlm.nih.gov/pubmed/36339591
http://dx.doi.org/10.3389/fphar.2022.1012622
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