A Sprouty4 reporter to monitor FGF/ERK signaling activity in ESCs and mice

The FGF/ERK signaling pathway is highly conserved throughout evolution and plays fundamental roles during embryonic development and in adult organisms. While a plethora of expression data exists for ligands, receptors and pathway regulators, we know little about the spatial organization or dynamics...

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Authors: Morgani, Sophie M., Saiz, Néstor, Garg, Vidur, Raina, Dhruv, Simon, Claire S., Kang, Minjung, Martinez Arias, Alfonso, Nichols, Jennifer, Schröter, Christian, Hadjantonakis, Anna-Katerina
Format: article
Status:Versión aceptada para publicación
Publication Date:2018
Country:España
Institution:Varias* (Consorci de Biblioteques Universitáries de Catalunya, Centre de Serveis Científics i Acadèmics de Catalunya)
Repository:Recercat. Dipósit de la Recerca de Catalunya
OAI Identifier:oai:recercat.cat:10230/56561
Online Access:http://hdl.handle.net/10230/56561
http://dx.doi.org/10.1016/j.ydbio.2018.06.017
Access Level:Open access
Keyword:ESCs
FGF signaling
FGF/ERK
Fluorescent reporter
Live imaging
Mouse embryo
Quantitative image analysis
Sprouty4
id ES_2c623aec73d0fcb2c84bb18bcd890b6e
oai_identifier_str oai:recercat.cat:10230/56561
network_acronym_str ES
network_name_str España
repository_id_str
spelling A Sprouty4 reporter to monitor FGF/ERK signaling activity in ESCs and miceMorgani, Sophie M.Saiz, NéstorGarg, VidurRaina, DhruvSimon, Claire S.Kang, MinjungMartinez Arias, AlfonsoNichols, JenniferSchröter, ChristianHadjantonakis, Anna-KaterinaESCsFGF signalingFGF/ERKFluorescent reporterLive imagingMouse embryoQuantitative image analysisSprouty4The FGF/ERK signaling pathway is highly conserved throughout evolution and plays fundamental roles during embryonic development and in adult organisms. While a plethora of expression data exists for ligands, receptors and pathway regulators, we know little about the spatial organization or dynamics of signaling in individual cells within populations. To this end we developed a transcriptional readout of FGF/ERK activity by targeting a histone H2B-linked Venus fluorophore to the endogenous locus of Spry4, an early pathway target, and generated Spry4H2B-Venus embryonic stem cells (ESCs) and a derivative mouse line. The Spry4H2B-Venus reporter was heterogeneously expressed within ESC cultures and responded to FGF/ERK signaling manipulation. In vivo, the Spry4H2B-Venus reporter recapitulated the expression pattern of Spry4 and localized to sites of known FGF/ERK activity including the inner cell mass of the pre-implantation embryo and the limb buds, somites and isthmus of the post-implantation embryo. Additionally, we observed highly localized reporter expression within adult organs. Genetic and chemical disruption of FGF/ERK signaling, in vivo in pre- and post-implantation embryos, abrogated Venus expression establishing the reporter as an accurate signaling readout. This tool will provide new insights into the dynamics of the FGF/ERK signaling pathway during mammalian development.Elsevier202320232018info:eu-repo/semantics/articleinfo:eu-repo/semantics/acceptedVersionapplication/pdfapplication/pdfhttp://hdl.handle.net/10230/56561http://dx.doi.org/10.1016/j.ydbio.2018.06.017reponame:Recercat. Dipósit de la Recerca de Catalunyainstname:Varias* (Consorci de Biblioteques Universitáries de Catalunya, Centre de Serveis Científics i Acadèmics de Catalunya)InglésDev Biol. 2018 Sep 1;441(1):104-26© Elsevier http://dx.doi.org/10.1016/j.ydbio.2018.06.017info:eu-repo/semantics/openAccessoai:recercat.cat:10230/565612026-05-29T05:05:01Z
dc.title.none.fl_str_mv A Sprouty4 reporter to monitor FGF/ERK signaling activity in ESCs and mice
title A Sprouty4 reporter to monitor FGF/ERK signaling activity in ESCs and mice
spellingShingle A Sprouty4 reporter to monitor FGF/ERK signaling activity in ESCs and mice
Morgani, Sophie M.
ESCs
FGF signaling
FGF/ERK
Fluorescent reporter
Live imaging
Mouse embryo
Quantitative image analysis
Sprouty4
title_short A Sprouty4 reporter to monitor FGF/ERK signaling activity in ESCs and mice
title_full A Sprouty4 reporter to monitor FGF/ERK signaling activity in ESCs and mice
title_fullStr A Sprouty4 reporter to monitor FGF/ERK signaling activity in ESCs and mice
title_full_unstemmed A Sprouty4 reporter to monitor FGF/ERK signaling activity in ESCs and mice
title_sort A Sprouty4 reporter to monitor FGF/ERK signaling activity in ESCs and mice
dc.creator.none.fl_str_mv Morgani, Sophie M.
Saiz, Néstor
Garg, Vidur
Raina, Dhruv
Simon, Claire S.
Kang, Minjung
Martinez Arias, Alfonso
Nichols, Jennifer
Schröter, Christian
Hadjantonakis, Anna-Katerina
author Morgani, Sophie M.
author_facet Morgani, Sophie M.
Saiz, Néstor
Garg, Vidur
Raina, Dhruv
Simon, Claire S.
Kang, Minjung
Martinez Arias, Alfonso
Nichols, Jennifer
Schröter, Christian
Hadjantonakis, Anna-Katerina
author_role author
author2 Saiz, Néstor
Garg, Vidur
Raina, Dhruv
Simon, Claire S.
Kang, Minjung
Martinez Arias, Alfonso
Nichols, Jennifer
Schröter, Christian
Hadjantonakis, Anna-Katerina
author2_role author
author
author
author
author
author
author
author
author
dc.subject.none.fl_str_mv ESCs
FGF signaling
FGF/ERK
Fluorescent reporter
Live imaging
Mouse embryo
Quantitative image analysis
Sprouty4
topic ESCs
FGF signaling
FGF/ERK
Fluorescent reporter
Live imaging
Mouse embryo
Quantitative image analysis
Sprouty4
description The FGF/ERK signaling pathway is highly conserved throughout evolution and plays fundamental roles during embryonic development and in adult organisms. While a plethora of expression data exists for ligands, receptors and pathway regulators, we know little about the spatial organization or dynamics of signaling in individual cells within populations. To this end we developed a transcriptional readout of FGF/ERK activity by targeting a histone H2B-linked Venus fluorophore to the endogenous locus of Spry4, an early pathway target, and generated Spry4H2B-Venus embryonic stem cells (ESCs) and a derivative mouse line. The Spry4H2B-Venus reporter was heterogeneously expressed within ESC cultures and responded to FGF/ERK signaling manipulation. In vivo, the Spry4H2B-Venus reporter recapitulated the expression pattern of Spry4 and localized to sites of known FGF/ERK activity including the inner cell mass of the pre-implantation embryo and the limb buds, somites and isthmus of the post-implantation embryo. Additionally, we observed highly localized reporter expression within adult organs. Genetic and chemical disruption of FGF/ERK signaling, in vivo in pre- and post-implantation embryos, abrogated Venus expression establishing the reporter as an accurate signaling readout. This tool will provide new insights into the dynamics of the FGF/ERK signaling pathway during mammalian development.
publishDate 2018
dc.date.none.fl_str_mv 2018
2023
2023
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/acceptedVersion
format article
status_str acceptedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/10230/56561
http://dx.doi.org/10.1016/j.ydbio.2018.06.017
url http://hdl.handle.net/10230/56561
http://dx.doi.org/10.1016/j.ydbio.2018.06.017
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.relation.none.fl_str_mv Dev Biol. 2018 Sep 1;441(1):104-26
dc.rights.none.fl_str_mv © Elsevier http://dx.doi.org/10.1016/j.ydbio.2018.06.017
info:eu-repo/semantics/openAccess
rights_invalid_str_mv © Elsevier http://dx.doi.org/10.1016/j.ydbio.2018.06.017
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
application/pdf
dc.publisher.none.fl_str_mv Elsevier
publisher.none.fl_str_mv Elsevier
dc.source.none.fl_str_mv reponame:Recercat. Dipósit de la Recerca de Catalunya
instname:Varias* (Consorci de Biblioteques Universitáries de Catalunya, Centre de Serveis Científics i Acadèmics de Catalunya)
instname_str Varias* (Consorci de Biblioteques Universitáries de Catalunya, Centre de Serveis Científics i Acadèmics de Catalunya)
reponame_str Recercat. Dipósit de la Recerca de Catalunya
collection Recercat. Dipósit de la Recerca de Catalunya
repository.name.fl_str_mv
repository.mail.fl_str_mv
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score 15,81155

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