A FRET-based cGMP biosensor in Drosophila

Gonzalez, Ana Clara - Abreu, Cecilia - Pantano, Sergio - Comini, Marcelo - Malacrida, Leonel - Egger, Boris - Cantera, Rafael - Prieto, Daniel

Resumen:

CUTie2 is a FRET-based cGMP biosensor tested so far only in cells. To expand its use to multicellular organisms we generated two transgenic Drosophila melanogaster strains that express the biosensor in a tissue-dependent manner. CUTie2 expression and subcellular localization was verified by confocal microscopy. The performance of CUTie2 was analyzed on dissected larval brains by hyperspectral microscopy and flow cytometry. Both approaches confirmed its responsivity, and the latter showed a rapid and reversible change in the fluorescence of the FRET acceptor upon cGMP treatment. This validated reporter system may prove valuable for studying cGMP signaling at organismal level.


Detalles Bibliográficos
2023
Agencia Nacional de Investigación e Innovación
Dirección Nacional de Innovación, Ciencia y Tecnología
Drosophila
FRET
cGMP
biosensor
Ciencias Naturales y Exactas
Ciencias Biológicas
Biología Celular, Microbiología
Inglés
Instituto de Investigaciones Biológicas Clemente Estable
IIBCE en REDI
https://hdl.handle.net/20.500.12381/3367
https://www.doi.org/10.17912/micropub.biology.000887
Acceso abierto
Reconocimiento 4.0 Internacional. (CC BY)
_version_ 1811155751578632192
author Gonzalez, Ana Clara
author2 Abreu, Cecilia
Pantano, Sergio
Comini, Marcelo
Malacrida, Leonel
Egger, Boris
Cantera, Rafael
Prieto, Daniel
author2_role author
author
author
author
author
author
author
author_facet Gonzalez, Ana Clara
Abreu, Cecilia
Pantano, Sergio
Comini, Marcelo
Malacrida, Leonel
Egger, Boris
Cantera, Rafael
Prieto, Daniel
author_role author
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MD5
bitstream.url.fl_str_mv https://redi.anii.org.uy/jspui/bitstream/20.500.12381/3367/2/license.txt
https://redi.anii.org.uy/jspui/bitstream/20.500.12381/3367/1/micropub-biology-000887%281%29.pdf
collection IIBCE en REDI
dc.creator.none.fl_str_mv Gonzalez, Ana Clara
Abreu, Cecilia
Pantano, Sergio
Comini, Marcelo
Malacrida, Leonel
Egger, Boris
Cantera, Rafael
Prieto, Daniel
dc.date.accessioned.none.fl_str_mv 2023-12-27T16:28:47Z
dc.date.available.none.fl_str_mv 2023-12-27T16:28:47Z
dc.date.issued.none.fl_str_mv 2023-11-28
dc.description.abstract.none.fl_txt_mv CUTie2 is a FRET-based cGMP biosensor tested so far only in cells. To expand its use to multicellular organisms we generated two transgenic Drosophila melanogaster strains that express the biosensor in a tissue-dependent manner. CUTie2 expression and subcellular localization was verified by confocal microscopy. The performance of CUTie2 was analyzed on dissected larval brains by hyperspectral microscopy and flow cytometry. Both approaches confirmed its responsivity, and the latter showed a rapid and reversible change in the fluorescence of the FRET acceptor upon cGMP treatment. This validated reporter system may prove valuable for studying cGMP signaling at organismal level.
dc.description.sponsorship.none.fl_txt_mv Agencia Nacional de Investigación e Innovación
Dirección Nacional de Innovación, Ciencia y Tecnología
dc.identifier.anii.es.fl_str_mv FCE_1_2019_1_156160
dc.identifier.doi.none.fl_str_mv https://www.doi.org/10.17912/micropub.biology.000887
dc.identifier.uri.none.fl_str_mv https://hdl.handle.net/20.500.12381/3367
dc.language.iso.none.fl_str_mv eng
dc.publisher.es.fl_str_mv micropublication.org
dc.rights.*.fl_str_mv Acceso abierto
dc.rights.license.none.fl_str_mv Reconocimiento 4.0 Internacional. (CC BY)
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
dc.source.es.fl_str_mv microPublication Biology
dc.source.none.fl_str_mv reponame:IIBCE en REDI
instname:Instituto de Investigaciones Biológicas Clemente Estable
instacron:Instituto de Investigaciones Biológicas Clemente Estable
dc.subject.anii.none.fl_str_mv Ciencias Naturales y Exactas
Ciencias Biológicas
Biología Celular, Microbiología
dc.subject.es.fl_str_mv Drosophila
FRET
cGMP
biosensor
dc.title.none.fl_str_mv A FRET-based cGMP biosensor in Drosophila
dc.type.es.fl_str_mv Artículo
dc.type.none.fl_str_mv info:eu-repo/semantics/article
dc.type.version.es.fl_str_mv Publicado
dc.type.version.none.fl_str_mv info:eu-repo/semantics/publishedVersion
description CUTie2 is a FRET-based cGMP biosensor tested so far only in cells. To expand its use to multicellular organisms we generated two transgenic Drosophila melanogaster strains that express the biosensor in a tissue-dependent manner. CUTie2 expression and subcellular localization was verified by confocal microscopy. The performance of CUTie2 was analyzed on dissected larval brains by hyperspectral microscopy and flow cytometry. Both approaches confirmed its responsivity, and the latter showed a rapid and reversible change in the fluorescence of the FRET acceptor upon cGMP treatment. This validated reporter system may prove valuable for studying cGMP signaling at organismal level.
eu_rights_str_mv openAccess
format article
id IIBCE_d30740091915b118f3a169ff592a32ae
identifier_str_mv FCE_1_2019_1_156160
instacron_str Instituto de Investigaciones Biológicas Clemente Estable
institution Instituto de Investigaciones Biológicas Clemente Estable
instname_str Instituto de Investigaciones Biológicas Clemente Estable
language eng
network_acronym_str IIBCE
network_name_str IIBCE en REDI
oai_identifier_str oai:redi.anii.org.uy:20.500.12381/3367
publishDate 2023
reponame_str IIBCE en REDI
repository.mail.fl_str_mv
repository.name.fl_str_mv IIBCE en REDI - Instituto de Investigaciones Biológicas Clemente Estable
repository_id_str 9421_3
rights_invalid_str_mv Reconocimiento 4.0 Internacional. (CC BY)
Acceso abierto
spelling Reconocimiento 4.0 Internacional. (CC BY)Acceso abiertoinfo:eu-repo/semantics/openAccess2023-12-27T16:28:47Z2023-12-27T16:28:47Z2023-11-28https://hdl.handle.net/20.500.12381/3367FCE_1_2019_1_156160https://www.doi.org/10.17912/micropub.biology.000887CUTie2 is a FRET-based cGMP biosensor tested so far only in cells. To expand its use to multicellular organisms we generated two transgenic Drosophila melanogaster strains that express the biosensor in a tissue-dependent manner. CUTie2 expression and subcellular localization was verified by confocal microscopy. The performance of CUTie2 was analyzed on dissected larval brains by hyperspectral microscopy and flow cytometry. Both approaches confirmed its responsivity, and the latter showed a rapid and reversible change in the fluorescence of the FRET acceptor upon cGMP treatment. This validated reporter system may prove valuable for studying cGMP signaling at organismal level.Agencia Nacional de Investigación e InnovaciónDirección Nacional de Innovación, Ciencia y Tecnologíaengmicropublication.orgmicroPublication Biologyreponame:IIBCE en REDIinstname:Instituto de Investigaciones Biológicas Clemente Estableinstacron:Instituto de Investigaciones Biológicas Clemente EstableDrosophilaFRETcGMPbiosensorCiencias Naturales y ExactasCiencias BiológicasBiología Celular, MicrobiologíaA FRET-based cGMP biosensor in DrosophilaArtículoPublicadoinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleInstituto de Investigaciones Biologicas Clemente EstableInstitut Pasteur de MontevideoUniversidad de la RepublicaUniverstity of Fribourg//Ciencias Naturales y Exactas/Ciencias Biológicas/Biología Celular, MicrobiologíaGonzalez, Ana ClaraAbreu, CeciliaPantano, SergioComini, MarceloMalacrida, LeonelEgger, BorisCantera, RafaelPrieto, DanielLICENSElicense.txtlicense.txttext/plain; charset=utf-85151https://redi.anii.org.uy/jspui/bitstream/20.500.12381/3367/2/license.txtfcc8b2ddb26c820405a37f9e0cae7f18MD52ORIGINALmicropub-biology-000887(1).pdfmicropub-biology-000887(1).pdfVersión publicada del artículoapplication/pdf1843581https://redi.anii.org.uy/jspui/bitstream/20.500.12381/3367/1/micropub-biology-000887%281%29.pdf0b69e90bdd5d8f4ac4cbc1b05d7062dbMD5120.500.12381/33672024-02-01 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://www.gub.uy/ministerio-educacion-cultura/iibcehttps://redi.anii.org.uy/oai/requestUruguayopendoar:9421_32024-02-01T17:14:32IIBCE 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spellingShingle A FRET-based cGMP biosensor in Drosophila
Gonzalez, Ana Clara
Drosophila
FRET
cGMP
biosensor
Ciencias Naturales y Exactas
Ciencias Biológicas
Biología Celular, Microbiología
status_str publishedVersion
title A FRET-based cGMP biosensor in Drosophila
title_full A FRET-based cGMP biosensor in Drosophila
title_fullStr A FRET-based cGMP biosensor in Drosophila
title_full_unstemmed A FRET-based cGMP biosensor in Drosophila
title_short A FRET-based cGMP biosensor in Drosophila
title_sort A FRET-based cGMP biosensor in Drosophila
topic Drosophila
FRET
cGMP
biosensor
Ciencias Naturales y Exactas
Ciencias Biológicas
Biología Celular, Microbiología
url https://hdl.handle.net/20.500.12381/3367
https://www.doi.org/10.17912/micropub.biology.000887