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

Journal of Visualized Experiments : JoVE
Anna Castells-NobauAnnette Schenck

Abstract

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 B...Continue Reading

Citations

May 20, 2020·Scientific Reports·Matthew J BroadheadGareth B Miles
Feb 13, 2019·PloS One·Anna Castells-NobauAnnette Schenck
May 16, 2019·Disease Models & Mechanisms·Mireia Coll-TanéAnnette Schenck
Feb 26, 2020·The Journal of Biological Chemistry·Villo MuhaDaan M F van Aalten

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Methods Mentioned

BETA
dissections
confocal microscopy
GTPase

Software Mentioned

NMJ
Drosophila
Fiji
_ Bouton
puncta
Drosophila NMJ Morphometrics
Analyze
Brp
NMJ Morphometrics
_ Morphometrics

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