NMJ analyser: a novel method to quantify neuromuscular junction morphology in zebrafish.

ABSTRACT:

Motivation

Neuromuscular junction structural integrity is crucial for transducing motoneuron signals that initiate skeletal muscle contraction. Zebrafish has emerged as a simple and efficient model to study neuromuscular junction structural morphology and function in the context of developmental neurobiology and neuromuscular diseases (NMDs). However, methods to quantify NMJ morphology from voluminous data of NMJ confocal images accurately, rapidly and reproducibly are lacking.

Results

We developed an ImageJ macro called "NMJ Analyser" to automatically and unbiasedly analyse NMJ morphology in zebrafish. From the Z-stack of a zebrafish hemi-somite, both presynaptic and postsynaptic fluorescently labelled termini at NMJs are extracted from background signal, with larger clusters of termini being segmented into individual termini using an unbiased algorithm. The program then determines whether each presynaptic terminus is colocalized with a postsynaptic terminus and vice versa, or whether it is orphaned, and tabulates the number of orphan and colocalized pre- and postsynaptic termini. The usefulness of this ImageJ macro plugin will be helpful to quantify NMJ parameters in zebrafish, particularly during development and in disease models of NMDs. It can enable high-throughput NMJ phenotypic screens in the drug discovery process for NMDs. It could also be further applied to the investigation of NMJ of other developmental systems.

Availability

NMJ Analyser is available for download at https://github.com/PattenLab/NMJ-Analyser.git.

Supplementary information

Supplementary data are available at Bioinformatics online.

SUBMITTER: Singh J 

PROVIDER: S-EPMC10713120 | biostudies-literature | 2023 Dec

REPOSITORIES: biostudies-literature