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Two Algorithms for High-throughput and Multi-parametric Quantification of Drosophila Neuromuscular Junction Morphology

Synaptic morphology is tightly related to synaptic efficacy, and in many cases morphological synapse defects ultimately lead to synaptic malfunction. The Drosophila larval neuromuscular junction (NMJ), a well-established model for glutamatergic synapses, has been extensively studied for decades. Ide...

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Autores principales: Castells-Nobau, Anna, Nijhof, Bonnie, Eidhof, Ilse, Wolf, Louis, Scheffer-de Gooyert, Jolanda M., Monedero, Ignacio, Torroja, Laura, van der Laak, Jeroen A.W.M., Schenck, Annette
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MyJove Corporation 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5607876/
https://www.ncbi.nlm.nih.gov/pubmed/28518121
http://dx.doi.org/10.3791/55395
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author Castells-Nobau, Anna
Nijhof, Bonnie
Eidhof, Ilse
Wolf, Louis
Scheffer-de Gooyert, Jolanda M.
Monedero, Ignacio
Torroja, Laura
van der Laak, Jeroen A.W.M.
Schenck, Annette
author_facet Castells-Nobau, Anna
Nijhof, Bonnie
Eidhof, Ilse
Wolf, Louis
Scheffer-de Gooyert, Jolanda M.
Monedero, Ignacio
Torroja, Laura
van der Laak, Jeroen A.W.M.
Schenck, Annette
author_sort Castells-Nobau, Anna
collection PubMed
description Synaptic morphology is tightly related to synaptic efficacy, and in many cases morphological synapse defects ultimately lead to synaptic malfunction. The Drosophila larval neuromuscular junction (NMJ), a well-established model for glutamatergic synapses, has been extensively studied for decades. Identification of mutations causing NMJ morphological defects revealed a repertoire of genes that regulate synapse development and function. Many of these were identified in large-scale studies that focused on qualitative approaches to detect morphological abnormalities of the Drosophila NMJ. A drawback of qualitative analyses is that many subtle players contributing to NMJ morphology likely remain unnoticed. Whereas quantitative analyses are required to detect the subtler morphological differences, such analyses are not yet commonly performed because they are laborious. This protocol describes in detail two image analysis algorithms "Drosophila NMJ Morphometrics" and "Drosophila NMJ Bouton Morphometrics", available as Fiji-compatible macros, for quantitative, accurate and objective morphometric analysis of the Drosophila NMJ. This methodology is developed to analyze NMJ terminals immunolabeled with the commonly used markers Dlg-1 and Brp. Additionally, its wider application to other markers such as Hrp, Csp and Syt is presented in this protocol. The macros are able to assess nine morphological NMJ features: NMJ area, NMJ perimeter, number of boutons, NMJ length, NMJ longest branch length, number of islands, number of branches, number of branching points and number of active zones in the NMJ terminal.
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spelling pubmed-56078762017-10-10 Two Algorithms for High-throughput and Multi-parametric Quantification of Drosophila Neuromuscular Junction Morphology Castells-Nobau, Anna Nijhof, Bonnie Eidhof, Ilse Wolf, Louis Scheffer-de Gooyert, Jolanda M. Monedero, Ignacio Torroja, Laura van der Laak, Jeroen A.W.M. Schenck, Annette J Vis Exp Neuroscience Synaptic morphology is tightly related to synaptic efficacy, and in many cases morphological synapse defects ultimately lead to synaptic malfunction. The Drosophila larval neuromuscular junction (NMJ), a well-established model for glutamatergic synapses, has been extensively studied for decades. Identification of mutations causing NMJ morphological defects revealed a repertoire of genes that regulate synapse development and function. Many of these were identified in large-scale studies that focused on qualitative approaches to detect morphological abnormalities of the Drosophila NMJ. A drawback of qualitative analyses is that many subtle players contributing to NMJ morphology likely remain unnoticed. Whereas quantitative analyses are required to detect the subtler morphological differences, such analyses are not yet commonly performed because they are laborious. This protocol describes in detail two image analysis algorithms "Drosophila NMJ Morphometrics" and "Drosophila NMJ Bouton Morphometrics", available as Fiji-compatible macros, for quantitative, accurate and objective morphometric analysis of the Drosophila NMJ. This methodology is developed to analyze NMJ terminals immunolabeled with the commonly used markers Dlg-1 and Brp. Additionally, its wider application to other markers such as Hrp, Csp and Syt is presented in this protocol. The macros are able to assess nine morphological NMJ features: NMJ area, NMJ perimeter, number of boutons, NMJ length, NMJ longest branch length, number of islands, number of branches, number of branching points and number of active zones in the NMJ terminal. MyJove Corporation 2017-05-03 /pmc/articles/PMC5607876/ /pubmed/28518121 http://dx.doi.org/10.3791/55395 Text en Copyright © 2017, Journal of Visualized Experiments http://creativecommons.org/licenses/by-nc-nd/3.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs 3.0 Unported License. To view a copy of this license, visithttp://creativecommons.org/licenses/by-nc-nd/3.0/
spellingShingle Neuroscience
Castells-Nobau, Anna
Nijhof, Bonnie
Eidhof, Ilse
Wolf, Louis
Scheffer-de Gooyert, Jolanda M.
Monedero, Ignacio
Torroja, Laura
van der Laak, Jeroen A.W.M.
Schenck, Annette
Two Algorithms for High-throughput and Multi-parametric Quantification of Drosophila Neuromuscular Junction Morphology
title Two Algorithms for High-throughput and Multi-parametric Quantification of Drosophila Neuromuscular Junction Morphology
title_full Two Algorithms for High-throughput and Multi-parametric Quantification of Drosophila Neuromuscular Junction Morphology
title_fullStr Two Algorithms for High-throughput and Multi-parametric Quantification of Drosophila Neuromuscular Junction Morphology
title_full_unstemmed Two Algorithms for High-throughput and Multi-parametric Quantification of Drosophila Neuromuscular Junction Morphology
title_short Two Algorithms for High-throughput and Multi-parametric Quantification of Drosophila Neuromuscular Junction Morphology
title_sort two algorithms for high-throughput and multi-parametric quantification of drosophila neuromuscular junction morphology
topic Neuroscience
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5607876/
https://www.ncbi.nlm.nih.gov/pubmed/28518121
http://dx.doi.org/10.3791/55395
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