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Two Algorithms for High-throughput and Multi-parametric Quantification of Drosophila Neuromuscular Junction Morphology
Radboud University of Nijmegen, Netherlands.
Radboud University of Nijmegen, Netherlands.
Radboud University of Nijmegen, Netherlands.
Radboud University of Nijmegen, Netherlands.
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2017 (English)In: Journal of Visualized Experiments, ISSN 1940-087X, E-ISSN 1940-087X, no 123, e55395Article in journal (Refereed) Published
Abstract [en]

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.

Place, publisher, year, edition, pages
JOURNAL OF VISUALIZED EXPERIMENTS , 2017. no 123, e55395
Keyword [en]
Neuroscience; Issue 123; Fiji; Drosophila; NMJ; macro; synapse; image segmentation; morphometry
National Category
Developmental Biology
Identifiers
URN: urn:nbn:se:liu:diva-139825DOI: 10.3791/55395ISI: 000406213900021PubMedID: 28518121OAI: oai:DiVA.org:liu-139825DiVA: diva2:1133895
Note

Funding Agencies|VIDI from Netherlands Organization for Scientific Research (NWO) [917-96-346, 912-12-109]; DCN/Radboud University Medical Center PhD fellowships; German Mental Retardation Network - NGFN+ program of the German Federal Ministry of Education and Research (BMBF); European Unions FP7 large scale integrated network Gencodys [HEALTH-241995]; TOP from Netherlands Organization for Scientific Research (NWO)

Available from: 2017-08-17 Created: 2017-08-17 Last updated: 2017-08-17

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Monedero Cobeta, Ignacio
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CiteExportLink to record
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Cite
Citation style
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