A New Fiji-Based Algorithm That Systematically Quantifies Nine Synaptic Parameters Provides Insights into Drosophila NMJ Morphometry

The morphology of synapses is of central interest in neuroscience because of the intimate relation with synaptic efficacy. Two decades of gene manipulation studies in different animal models have revealed a repertoire of molecules that contribute to synapse development. However, since such studies o...

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Autores: Nijhof, B., Castells-Nobau, A., Wolf, L., Scheffer-de Gooyert, J.M., Monedero Cobeta, Ignacio, Torroja Fungairiño, Laura, Coromina, L., Van der Laak, J.A.W.M., Schenck, A.
Tipo de recurso: artículo
Fecha de publicación:2016
País:España
Institución:Universidad Autónoma de Madrid
Repositorio:Biblos-e Archivo. Repositorio Institucional de la UAM
Idioma:inglés
OAI Identifier:oai:repositorio.uam.es:10486/679222
Acceso en línea:http://hdl.handle.net/10486/679222
https://dx.doi.org/10.1371/journal.pcbi.1004823
Access Level:acceso abierto
Palabra clave:Drosophila
Fiji
Gender
Genetic background
Geometry
Biología y Biomedicina / Biología
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spelling A New Fiji-Based Algorithm That Systematically Quantifies Nine Synaptic Parameters Provides Insights into Drosophila NMJ MorphometryNijhof, B.Castells-Nobau, A.Wolf, L.Scheffer-de Gooyert, J.M.Monedero Cobeta, IgnacioTorroja Fungairiño, LauraCoromina, L.Van der Laak, J.A.W.M.Schenck, A.DrosophilaFijiGenderGenetic backgroundGeometryBiología y Biomedicina / BiologíaThe morphology of synapses is of central interest in neuroscience because of the intimate relation with synaptic efficacy. Two decades of gene manipulation studies in different animal models have revealed a repertoire of molecules that contribute to synapse development. However, since such studies often assessed only one, or at best a few, morphological features at a given synapse, it remained unaddressed how different structural aspects relate to one another. Furthermore, such focused and sometimes only qualitative approaches likely left many of the more subtle players unnoticed. Here, we present the image analysis algorithm ‘Drosophila_NMJ_Morphometrics’, available as a Fiji-compatible macro, for quantitative, accurate and objective synapse morphometry of the Drosophila larval neuromuscular junction (NMJ), a well-established glutamatergic model synapse. We developed this methodology for semi-automated multiparametric analyses of NMJ terminals immunolabeled for the commonly used markers Dlg1 and Brp and showed that it also works for Hrp, Csp and Syt. We demonstrate that gender, genetic background and identity of abdominal body segment consistently and significantly contribute to variability in our data, suggesting that controlling for these parameters is important to minimize variability in quantitative analyses. Correlation and principal component analyses (PCA) were performed to investigate which morphometric parameters are inter-dependent and which ones are regulated rather independently. Based on nine acquired parameters, we identified five morphometric groups: NMJ size, geometry, muscle size, number of NMJ islands and number of active zones. Based on our finding that the parameters of the first two principal components hardly correlated with each other, we suggest that different molecular processes underlie these two morphometric groups. Our study sets the stage for systems morphometry approaches at the well-studied Drosophila NMJThis study was supported by VIDI and TOP grants (917-96-346, 912-12-109) from the Netherlands Organization for Scientific Research (NWO), by a DCN/Radboud University Medical Center PhD fellowship, by the German Mental Retardation Network funded by the NGFN+ program of the German Federal Ministry of Education and Research (BMBF) and by the European Union's FP7 large scale integrated network Gencodys (HEALTH-241995) to ASPublic Library of ScienceDepartamento de BiologíaFacultad de Ciencias20162016-03-21research articlehttp://purl.org/coar/resource_type/c_2df8fbb1VoRhttp://purl.org/coar/version/c_970fb48d4fbd8a85info:eu-repo/semantics/articleapplication/pdfhttp://hdl.handle.net/10486/679222https://dx.doi.org/10.1371/journal.pcbi.1004823reponame:Biblos-e Archivo. Repositorio Institucional de la UAMinstname:Universidad Autónoma de MadridInglésengEuropean Commission http://dx.doi.org/10.13039/501100000780 Framework Programme Seven 241995open accesshttp://purl.org/coar/access_right/c_abf2info:eu-repo/semantics/openAccessoai:repositorio.uam.es:10486/6792222026-06-23T12:46:27Z
dc.title.none.fl_str_mv A New Fiji-Based Algorithm That Systematically Quantifies Nine Synaptic Parameters Provides Insights into Drosophila NMJ Morphometry
title A New Fiji-Based Algorithm That Systematically Quantifies Nine Synaptic Parameters Provides Insights into Drosophila NMJ Morphometry
spellingShingle A New Fiji-Based Algorithm That Systematically Quantifies Nine Synaptic Parameters Provides Insights into Drosophila NMJ Morphometry
Nijhof, B.
Drosophila
Fiji
Gender
Genetic background
Geometry
Biología y Biomedicina / Biología
title_short A New Fiji-Based Algorithm That Systematically Quantifies Nine Synaptic Parameters Provides Insights into Drosophila NMJ Morphometry
title_full A New Fiji-Based Algorithm That Systematically Quantifies Nine Synaptic Parameters Provides Insights into Drosophila NMJ Morphometry
title_fullStr A New Fiji-Based Algorithm That Systematically Quantifies Nine Synaptic Parameters Provides Insights into Drosophila NMJ Morphometry
title_full_unstemmed A New Fiji-Based Algorithm That Systematically Quantifies Nine Synaptic Parameters Provides Insights into Drosophila NMJ Morphometry
title_sort A New Fiji-Based Algorithm That Systematically Quantifies Nine Synaptic Parameters Provides Insights into Drosophila NMJ Morphometry
dc.creator.none.fl_str_mv Nijhof, B.
Castells-Nobau, A.
Wolf, L.
Scheffer-de Gooyert, J.M.
Monedero Cobeta, Ignacio
Torroja Fungairiño, Laura
Coromina, L.
Van der Laak, J.A.W.M.
Schenck, A.
author Nijhof, B.
author_facet Nijhof, B.
Castells-Nobau, A.
Wolf, L.
Scheffer-de Gooyert, J.M.
Monedero Cobeta, Ignacio
Torroja Fungairiño, Laura
Coromina, L.
Van der Laak, J.A.W.M.
Schenck, A.
author_role author
author2 Castells-Nobau, A.
Wolf, L.
Scheffer-de Gooyert, J.M.
Monedero Cobeta, Ignacio
Torroja Fungairiño, Laura
Coromina, L.
Van der Laak, J.A.W.M.
Schenck, A.
author2_role author
author
author
author
author
author
author
author
dc.contributor.none.fl_str_mv Departamento de Biología
Facultad de Ciencias
dc.subject.none.fl_str_mv Drosophila
Fiji
Gender
Genetic background
Geometry
Biología y Biomedicina / Biología
topic Drosophila
Fiji
Gender
Genetic background
Geometry
Biología y Biomedicina / Biología
description The morphology of synapses is of central interest in neuroscience because of the intimate relation with synaptic efficacy. Two decades of gene manipulation studies in different animal models have revealed a repertoire of molecules that contribute to synapse development. However, since such studies often assessed only one, or at best a few, morphological features at a given synapse, it remained unaddressed how different structural aspects relate to one another. Furthermore, such focused and sometimes only qualitative approaches likely left many of the more subtle players unnoticed. Here, we present the image analysis algorithm ‘Drosophila_NMJ_Morphometrics’, available as a Fiji-compatible macro, for quantitative, accurate and objective synapse morphometry of the Drosophila larval neuromuscular junction (NMJ), a well-established glutamatergic model synapse. We developed this methodology for semi-automated multiparametric analyses of NMJ terminals immunolabeled for the commonly used markers Dlg1 and Brp and showed that it also works for Hrp, Csp and Syt. We demonstrate that gender, genetic background and identity of abdominal body segment consistently and significantly contribute to variability in our data, suggesting that controlling for these parameters is important to minimize variability in quantitative analyses. Correlation and principal component analyses (PCA) were performed to investigate which morphometric parameters are inter-dependent and which ones are regulated rather independently. Based on nine acquired parameters, we identified five morphometric groups: NMJ size, geometry, muscle size, number of NMJ islands and number of active zones. Based on our finding that the parameters of the first two principal components hardly correlated with each other, we suggest that different molecular processes underlie these two morphometric groups. Our study sets the stage for systems morphometry approaches at the well-studied Drosophila NMJ
publishDate 2016
dc.date.none.fl_str_mv 2016
2016-03-21
dc.type.none.fl_str_mv research article
http://purl.org/coar/resource_type/c_2df8fbb1
VoR
http://purl.org/coar/version/c_970fb48d4fbd8a85
dc.type.openaire.fl_str_mv info:eu-repo/semantics/article
format article
dc.identifier.none.fl_str_mv http://hdl.handle.net/10486/679222
https://dx.doi.org/10.1371/journal.pcbi.1004823
url http://hdl.handle.net/10486/679222
https://dx.doi.org/10.1371/journal.pcbi.1004823
dc.language.none.fl_str_mv Inglés
eng
language_invalid_str_mv Inglés
language eng
dc.relation.none.fl_str_mv European Commission http://dx.doi.org/10.13039/501100000780 Framework Programme Seven 241995
dc.rights.none.fl_str_mv open access
http://purl.org/coar/access_right/c_abf2
dc.rights.openaire.fl_str_mv info:eu-repo/semantics/openAccess
rights_invalid_str_mv open access
http://purl.org/coar/access_right/c_abf2
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv Public Library of Science
publisher.none.fl_str_mv Public Library of Science
dc.source.none.fl_str_mv reponame:Biblos-e Archivo. Repositorio Institucional de la UAM
instname:Universidad Autónoma de Madrid
instname_str Universidad Autónoma de Madrid
reponame_str Biblos-e Archivo. Repositorio Institucional de la UAM
collection Biblos-e Archivo. Repositorio Institucional de la UAM
repository.name.fl_str_mv
repository.mail.fl_str_mv
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