Reversible silencing of endogenous receptors in intact brain tissue using 2-photon pharmacology

The physiological activity of proteins is often studied with loss-of-function genetic approaches, but the corresponding phenotypes develop slowly and can be confounding. Photopharmacology allows direct, fast, and reversible control of endogenous protein activity, with spatiotemporal resolution set b...

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Autores: Pittolo, Silvia, Lee, Hyojung, Lladó, Anna, Tosi, Sébastien, Bosch, Miquel, Bardia, Lídia, Gómez-Santacana, Xavier, Llebaria, Amadeu, Soriano, Eduardo, Colombelli, Julien, Poskanzer, Kira E., Perea, Gertrudis, Gorostiza, Pau
Tipo de documento: artigo
Estado:Versão publicada
Data de publicação:2019
País:España
Recursos:Consejo Superior de Investigaciones Científicas (CSIC)
Repositório:DIGITAL.CSIC. Repositorio Institucional del CSIC
OAI Identifier:oai:digital.csic.es:10261/200686
Acesso em linha:http://hdl.handle.net/10261/200686
Access Level:Acceso aberto
Palavra-chave:2-photon pharmacology
Functional silencing
Pharmacological selectivity
Photoactivation
Photopharmacology
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spelling Reversible silencing of endogenous receptors in intact brain tissue using 2-photon pharmacologyPittolo, SilviaLee, HyojungLladó, AnnaTosi, SébastienBosch, MiquelBardia, LídiaGómez-Santacana, XavierLlebaria, AmadeuSoriano, EduardoColombelli, JulienPoskanzer, Kira E.Perea, GertrudisGorostiza, Pau2-photon pharmacologyFunctional silencingPharmacological selectivityPhotoactivationPhotopharmacologyThe physiological activity of proteins is often studied with loss-of-function genetic approaches, but the corresponding phenotypes develop slowly and can be confounding. Photopharmacology allows direct, fast, and reversible control of endogenous protein activity, with spatiotemporal resolution set by the illumination method. Here, we combine a photoswitchable allosteric modulator (alloswitch) and 2-photon excitation using pulsed near-infrared lasers to reversibly silence metabotropic glutamate 5 (mGlu5) receptor activity in intact brain tissue. Endogenous receptors can be photoactivated in neurons and astrocytes with pharmacological selectivity and with an axial resolution between 5 and 10 μm. Thus, 2-photon pharmacology using alloswitch allows investigating mGlu5-dependent processes in wild-type animals, including synaptic formation and plasticity, and signaling pathways from intracellular organelles. © 2019 National Academy of Sciences. All rights reserved.ACKNOWLEDGMENTS. We thank Jordi Hernando (Autonomous University of Barcelona) for useful discussions on 2-photon excitation; Pere Català (Utrecht University) for help with GCaMP; Francisco Ciruela (University of Barcelona) for mGlu5-eYFP plasmid; Erin Schuman and Stephan Junek (Max Planck Institute for Brain Research, Frankfurt) for preliminary 2-photon excitation experiments; and Ashraf Muhaisen (University of Barcelona) for help with slicing. This research received funding from European Union Research and Innovation Programme Horizon 2020 [Human Brain Project SGA2 Grant Agreement 785907 (WaveScalES)], European Research ERA-Net SynBio programme (Modulightor project), and financial support from Agency for Management of University and Research Grants/Generalitat de Catalunya (CERCA Programme; 2017-SGR-1442 project), Fonds Européen de Développement Économique et Régional (FEDER) funds, Ministry of Economy and Competitiveness (MINECO)/FEDER (Grant CTQ2016-80066-R), and the Fundaluce foundation. S.P. was supported by an FI fellowship from the Agency for Management of University and Research Grants/Generalitat de Catalunya (2014FI_B2 00160). H.L. was supported by an Institute for Bioengineering of Catalonia Severo Ochoa International PhD Programme fellowship from MINECO. M.B. was supported by a H2020-MSCA-IF Reintegration Grant. K.E.P. receives support from NIH/National Institute of Neurological Disorders and Stroke Grant R01NS099254 and NSF Biophotonics Grant 1604544. E.S. receives support from MINECO (Grant SAF2016-7426).Peer reviewedNational Academy of Sciences (U.S.)European CommissionMinisterio de Economía y Competitividad (España)Llebaría, Amadeu [0000-0002-8200-4827]Consejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72]202020202019info:eu-repo/semantics/articlehttp://purl.org/coar/resource_type/c_6501Publisher's versioninfo:eu-repo/semantics/publishedVersionhttp://hdl.handle.net/10261/200686reponame:DIGITAL.CSIC. Repositorio Institucional del CSICinstname:Consejo Superior de Investigaciones Científicas (CSIC)Inglés#PLACEHOLDER_PARENT_METADATA_VALUE##PLACEHOLDER_PARENT_METADATA_VALUE##PLACEHOLDER_PARENT_METADATA_VALUE#info:eu-repo/grantAgreement/EC/H2020/785907info:eu-repo/grantAgreement/MINECO/Plan Estatal de Investigación Científica y Técnica y de Innovación 2013-2016/CTQ2016-80066-Rinfo:eu-repo/grantAgreement/MINECO/Plan Estatal de Investigación Científica y Técnica y de Innovación 2013-2016/SAF2016-7426-Rhttps://doi.org/10.1073/pnas.1900430116Síinfo:eu-repo/semantics/openAccessoai:digital.csic.es:10261/2006862026-05-22T06:33:51Z
dc.title.none.fl_str_mv Reversible silencing of endogenous receptors in intact brain tissue using 2-photon pharmacology
title Reversible silencing of endogenous receptors in intact brain tissue using 2-photon pharmacology
spellingShingle Reversible silencing of endogenous receptors in intact brain tissue using 2-photon pharmacology
Pittolo, Silvia
2-photon pharmacology
Functional silencing
Pharmacological selectivity
Photoactivation
Photopharmacology
title_short Reversible silencing of endogenous receptors in intact brain tissue using 2-photon pharmacology
title_full Reversible silencing of endogenous receptors in intact brain tissue using 2-photon pharmacology
title_fullStr Reversible silencing of endogenous receptors in intact brain tissue using 2-photon pharmacology
title_full_unstemmed Reversible silencing of endogenous receptors in intact brain tissue using 2-photon pharmacology
title_sort Reversible silencing of endogenous receptors in intact brain tissue using 2-photon pharmacology
dc.creator.none.fl_str_mv Pittolo, Silvia
Lee, Hyojung
Lladó, Anna
Tosi, Sébastien
Bosch, Miquel
Bardia, Lídia
Gómez-Santacana, Xavier
Llebaria, Amadeu
Soriano, Eduardo
Colombelli, Julien
Poskanzer, Kira E.
Perea, Gertrudis
Gorostiza, Pau
author Pittolo, Silvia
author_facet Pittolo, Silvia
Lee, Hyojung
Lladó, Anna
Tosi, Sébastien
Bosch, Miquel
Bardia, Lídia
Gómez-Santacana, Xavier
Llebaria, Amadeu
Soriano, Eduardo
Colombelli, Julien
Poskanzer, Kira E.
Perea, Gertrudis
Gorostiza, Pau
author_role author
author2 Lee, Hyojung
Lladó, Anna
Tosi, Sébastien
Bosch, Miquel
Bardia, Lídia
Gómez-Santacana, Xavier
Llebaria, Amadeu
Soriano, Eduardo
Colombelli, Julien
Poskanzer, Kira E.
Perea, Gertrudis
Gorostiza, Pau
author2_role author
author
author
author
author
author
author
author
author
author
author
author
dc.contributor.none.fl_str_mv European Commission
Ministerio de Economía y Competitividad (España)
Llebaría, Amadeu [0000-0002-8200-4827]
Consejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72]
dc.subject.none.fl_str_mv 2-photon pharmacology
Functional silencing
Pharmacological selectivity
Photoactivation
Photopharmacology
topic 2-photon pharmacology
Functional silencing
Pharmacological selectivity
Photoactivation
Photopharmacology
description The physiological activity of proteins is often studied with loss-of-function genetic approaches, but the corresponding phenotypes develop slowly and can be confounding. Photopharmacology allows direct, fast, and reversible control of endogenous protein activity, with spatiotemporal resolution set by the illumination method. Here, we combine a photoswitchable allosteric modulator (alloswitch) and 2-photon excitation using pulsed near-infrared lasers to reversibly silence metabotropic glutamate 5 (mGlu5) receptor activity in intact brain tissue. Endogenous receptors can be photoactivated in neurons and astrocytes with pharmacological selectivity and with an axial resolution between 5 and 10 μm. Thus, 2-photon pharmacology using alloswitch allows investigating mGlu5-dependent processes in wild-type animals, including synaptic formation and plasticity, and signaling pathways from intracellular organelles. © 2019 National Academy of Sciences. All rights reserved.
publishDate 2019
dc.date.none.fl_str_mv 2019
2020
2020
dc.type.none.fl_str_mv info:eu-repo/semantics/article
http://purl.org/coar/resource_type/c_6501
Publisher's version
info:eu-repo/semantics/publishedVersion
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/10261/200686
url http://hdl.handle.net/10261/200686
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.relation.none.fl_str_mv #PLACEHOLDER_PARENT_METADATA_VALUE#
#PLACEHOLDER_PARENT_METADATA_VALUE#
#PLACEHOLDER_PARENT_METADATA_VALUE#
info:eu-repo/grantAgreement/EC/H2020/785907
info:eu-repo/grantAgreement/MINECO/Plan Estatal de Investigación Científica y Técnica y de Innovación 2013-2016/CTQ2016-80066-R
info:eu-repo/grantAgreement/MINECO/Plan Estatal de Investigación Científica y Técnica y de Innovación 2013-2016/SAF2016-7426-R
https://doi.org/10.1073/pnas.1900430116
Sí
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.publisher.none.fl_str_mv National Academy of Sciences (U.S.)
publisher.none.fl_str_mv National Academy of Sciences (U.S.)
dc.source.none.fl_str_mv reponame:DIGITAL.CSIC. Repositorio Institucional del CSIC
instname:Consejo Superior de Investigaciones Científicas (CSIC)
instname_str Consejo Superior de Investigaciones Científicas (CSIC)
reponame_str DIGITAL.CSIC. Repositorio Institucional del CSIC
collection DIGITAL.CSIC. Repositorio Institucional del CSIC
repository.name.fl_str_mv
repository.mail.fl_str_mv
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score 15,812429

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