Simultaneous detection of membrane contact dynamics and associated Ca2+ signals by reversible chemogenetic reporters
Membrane contact sites (MCSs) are hubs allowing various cell organelles to coordinate their activities. The dynamic nature of these sites and their small size hinder analysis by current imaging techniques. To overcome these limitations, we here design a series of reversible chemogenetic reporters in...
| Autores: | , , , , , , , , , , , , , , , , , , , , , , |
|---|---|
| Tipo de recurso: | artículo |
| Estado: | Versión publicada |
| Fecha de publicación: | 2024 |
| País: | España |
| Institución: | Consejo Superior de Investigaciones Científicas (CSIC) |
| Repositorio: | DIGITAL.CSIC. Repositorio Institucional del CSIC |
| OAI Identifier: | oai:digital.csic.es:10261/380317 |
| Acceso en línea: | http://hdl.handle.net/10261/380317 |
| Access Level: | acceso abierto |
| Palabra clave: | Ca2+ imaging Calcium signalling Endoplasmic reticulum Fluorescence imaging Fluorescent proteins |
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| dc.title.none.fl_str_mv |
Simultaneous detection of membrane contact dynamics and associated Ca2+ signals by reversible chemogenetic reporters |
| title |
Simultaneous detection of membrane contact dynamics and associated Ca2+ signals by reversible chemogenetic reporters |
| spellingShingle |
Simultaneous detection of membrane contact dynamics and associated Ca2+ signals by reversible chemogenetic reporters García-Casas, Paloma Ca2+ imaging Calcium signalling Endoplasmic reticulum Fluorescence imaging Fluorescent proteins |
| title_short |
Simultaneous detection of membrane contact dynamics and associated Ca2+ signals by reversible chemogenetic reporters |
| title_full |
Simultaneous detection of membrane contact dynamics and associated Ca2+ signals by reversible chemogenetic reporters |
| title_fullStr |
Simultaneous detection of membrane contact dynamics and associated Ca2+ signals by reversible chemogenetic reporters |
| title_full_unstemmed |
Simultaneous detection of membrane contact dynamics and associated Ca2+ signals by reversible chemogenetic reporters |
| title_sort |
Simultaneous detection of membrane contact dynamics and associated Ca2+ signals by reversible chemogenetic reporters |
| dc.creator.none.fl_str_mv |
García-Casas, Paloma Rossini, Michela Påvénius, Linnea Saeed, Mezida Arnst, Nikita Sonda, Sonia Fernandes, Tânia D’Arsiè, Irene Bruzzone, Matteo Berno, Valeria Raimondi, Andrea Livia Sassano, Maria Naia, Luana Barbieri, Elisa Sigismund, Sara Agostinis, Patrizia Sturlese, Mattia Niemeyer, Barbara A. Brismar, Hjalmar Ankarcrona, María Gautier, Arnaud Pizzo, Paola Filadi, Riccardo |
| author |
García-Casas, Paloma |
| author_facet |
García-Casas, Paloma Rossini, Michela Påvénius, Linnea Saeed, Mezida Arnst, Nikita Sonda, Sonia Fernandes, Tânia D’Arsiè, Irene Bruzzone, Matteo Berno, Valeria Raimondi, Andrea Livia Sassano, Maria Naia, Luana Barbieri, Elisa Sigismund, Sara Agostinis, Patrizia Sturlese, Mattia Niemeyer, Barbara A. Brismar, Hjalmar Ankarcrona, María Gautier, Arnaud Pizzo, Paola Filadi, Riccardo |
| author_role |
author |
| author2 |
Rossini, Michela Påvénius, Linnea Saeed, Mezida Arnst, Nikita Sonda, Sonia Fernandes, Tânia D’Arsiè, Irene Bruzzone, Matteo Berno, Valeria Raimondi, Andrea Livia Sassano, Maria Naia, Luana Barbieri, Elisa Sigismund, Sara Agostinis, Patrizia Sturlese, Mattia Niemeyer, Barbara A. Brismar, Hjalmar Ankarcrona, María Gautier, Arnaud Pizzo, Paola Filadi, Riccardo |
| author2_role |
author author author author author author author author author author author author author author author author author author author author author author |
| dc.contributor.none.fl_str_mv |
Universidad de Valladolid Ministerio de Educación (España) European Commission Associazione Italiana per la Ricerca sul Cancro Ministero dell'Istruzione, dell'Università e della Ricerca Università degli Studi di Padova Chan Zuckerberg Initiative Silicon Valley Community Foundation Consejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72] |
| dc.subject.none.fl_str_mv |
Ca2+ imaging Calcium signalling Endoplasmic reticulum Fluorescence imaging Fluorescent proteins |
| topic |
Ca2+ imaging Calcium signalling Endoplasmic reticulum Fluorescence imaging Fluorescent proteins |
| description |
Membrane contact sites (MCSs) are hubs allowing various cell organelles to coordinate their activities. The dynamic nature of these sites and their small size hinder analysis by current imaging techniques. To overcome these limitations, we here design a series of reversible chemogenetic reporters incorporating improved, low-affinity variants of splitFAST, and study the dynamics of different MCSs at high spatiotemporal resolution, both in vitro and in vivo. We demonstrate that these versatile reporters suit different experimental setups well, allowing one to address challenging biological questions. Using these probes, we identify a pathway in which calcium (Ca2+) signalling dynamically regulates endoplasmic reticulum-mitochondria juxtaposition, characterizing the underlying mechanism. Finally, by integrating Ca2+-sensing capabilities into the splitFAST technology, we introduce PRINCESS (PRobe for INterorganelle Ca2+-Exchange Sites based on SplitFAST), a class of reporters to simultaneously detect MCSs and measure the associated Ca2+ dynamics using a single biosensor. |
| publishDate |
2024 |
| dc.date.none.fl_str_mv |
2024 2025 2025 |
| 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 |
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article |
| status_str |
publishedVersion |
| dc.identifier.none.fl_str_mv |
http://hdl.handle.net/10261/380317 |
| url |
http://hdl.handle.net/10261/380317 |
| dc.language.none.fl_str_mv |
Inglés |
| language_invalid_str_mv |
Inglés |
| dc.relation.none.fl_str_mv |
#PLACEHOLDER_PARENT_METADATA_VALUE# info:eu-repo/grantAgreement/EC/H2020/101002280 The underlying dataset has been published as supplementary material of the article in the publisher platform at DOI https://doi.org/10.1038/s41467-024-52985-0 https://doi.org/10.1038/s41467-024-52985-0 Sí |
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info:eu-repo/semantics/openAccess |
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openAccess |
| dc.format.none.fl_str_mv |
application/pdf |
| dc.publisher.none.fl_str_mv |
Springer Nature |
| publisher.none.fl_str_mv |
Springer Nature |
| dc.source.none.fl_str_mv |
reponame:DIGITAL.CSIC. Repositorio Institucional del CSIC instname:Consejo Superior de Investigaciones Científicas (CSIC) |
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Consejo Superior de Investigaciones Científicas (CSIC) |
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DIGITAL.CSIC. Repositorio Institucional del CSIC |
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DIGITAL.CSIC. Repositorio Institucional del CSIC |
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| _version_ |
1869413475586932736 |
| spelling |
Simultaneous detection of membrane contact dynamics and associated Ca2+ signals by reversible chemogenetic reportersGarcía-Casas, PalomaRossini, MichelaPåvénius, LinneaSaeed, MezidaArnst, NikitaSonda, SoniaFernandes, TâniaD’Arsiè, IreneBruzzone, MatteoBerno, ValeriaRaimondi, AndreaLivia Sassano, MariaNaia, LuanaBarbieri, ElisaSigismund, SaraAgostinis, PatriziaSturlese, MattiaNiemeyer, Barbara A.Brismar, HjalmarAnkarcrona, MaríaGautier, ArnaudPizzo, PaolaFiladi, RiccardoCa2+ imagingCalcium signallingEndoplasmic reticulumFluorescence imagingFluorescent proteinsMembrane contact sites (MCSs) are hubs allowing various cell organelles to coordinate their activities. The dynamic nature of these sites and their small size hinder analysis by current imaging techniques. To overcome these limitations, we here design a series of reversible chemogenetic reporters incorporating improved, low-affinity variants of splitFAST, and study the dynamics of different MCSs at high spatiotemporal resolution, both in vitro and in vivo. We demonstrate that these versatile reporters suit different experimental setups well, allowing one to address challenging biological questions. Using these probes, we identify a pathway in which calcium (Ca2+) signalling dynamically regulates endoplasmic reticulum-mitochondria juxtaposition, characterizing the underlying mechanism. Finally, by integrating Ca2+-sensing capabilities into the splitFAST technology, we introduce PRINCESS (PRobe for INterorganelle Ca2+-Exchange Sites based on SplitFAST), a class of reporters to simultaneously detect MCSs and measure the associated Ca2+ dynamics using a single biosensor.The authors gratefully acknowledge L. Ozmen and F. Hoffmann-La Roche Ltd (Basel, Switzerland) for kindly donating the AD (B6.152H) mouse model used in this study (MTA 02-01-2022-UniPD), E. Trevisson, V. Morbidoni (University of Padua) and A. Miranda Vizuete for helping with the setting of the C. elegans strains, P. Romani for helping with the setting of confocal microscopy, D. Sandonà for suggestions on cloning strategies, I. Costa and M. Gintoli for training and suggestions on FLIM acquisitions and P. Magalhães for proofreading of the manuscript. Part of the schemes presented in this work were created with BioRender.com. P.G.C fellowship was supported by Ayudas de recualificación del Sistema Universitario Margarita Salas (2021–2023), University of Valladolid-Spanish Ministry of Education, financed by the European Union through EU Next Generation. T.F. fellowship was supported by PRIN 2022943TH9. S.S. was supported by grants from the European Research Council (ERC-CoG2020 101002280) and Associazione Italiana per la Ricerca sul Cancro (AIRC IG 24415). This work was supported by grants from the Italian Ministry of University and Scientific Research (PRIN2017XA5J5N and PRIN2022943TH9, financed by the EU, NextGenerationEU); the University of Padova (SID2019); Cure Alzheimer’s Fund (USA) to P.P.; the Dynamic Imaging program of the Chan-Zuckerberg Initiative DAF (grant number 2023-321185), an advised fund of Silicon Valley Community Foundation, to A.G. and R.F; the Italian Ministry of University and Scientific Research grant (PRIN P20225R4Y5, financed by the European Union, NextGenerationEU) to P.P. and R.F. The authors acknowledge Euro-BioImaging [https://www.eurobioimaging.eu/] for providing access to imaging technologies and services via the Advanced Light Microscopy Italian Node (Laboratory of Ca2+ and cAMP signaling in physiology and pathology, Padua, Italy; and ALEMBIC, Milan, Italy); the Swedish National Microscopy Infrastructure, NMI (VR-RFI 2019-00217); EuroBioImaging-ERIC and the PNRR infrastructure, SEELIFE n. IR00023 (financed by the European Union, NextGenerationEU, Missione 4, Componente 2, CUP B53C22001810006).Peer reviewedSpringer NatureUniversidad de ValladolidMinisterio de Educación (España)European CommissionAssociazione Italiana per la Ricerca sul CancroMinistero dell'Istruzione, dell'Università e della RicercaUniversità degli Studi di PadovaChan Zuckerberg InitiativeSilicon Valley Community FoundationConsejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72]202520252024info:eu-repo/semantics/articlehttp://purl.org/coar/resource_type/c_6501Publisher's versioninfo:eu-repo/semantics/publishedVersionapplication/pdfhttp://hdl.handle.net/10261/380317reponame:DIGITAL.CSIC. Repositorio Institucional del CSICinstname:Consejo Superior de Investigaciones Científicas (CSIC)Inglés#PLACEHOLDER_PARENT_METADATA_VALUE#info:eu-repo/grantAgreement/EC/H2020/101002280The underlying dataset has been published as supplementary material of the article in the publisher platform at DOI https://doi.org/10.1038/s41467-024-52985-0https://doi.org/10.1038/s41467-024-52985-0Síinfo:eu-repo/semantics/openAccessoai:digital.csic.es:10261/3803172026-05-22T06:33:51Z |
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15.812429 |