ANMJ-morph: a simple macro for rapid analysis of neuromuscular junction morphology. Issue 4 (15th April 2020)

• Record Type: Journal Article
• Authors: Minty, Gavin; Hoppen, Alex; Boehm, Ines; Alhindi, Abrar; Gibb, Larissa; Potter, Ellie; Wagner, Boris C.; Miller, Janice; Skipworth, Richard J. E.; Gillingwater, Thomas H.; Jones, Ross A.
• Is Part Of: Royal Society open science. Volume 7:Issue 4(2020)
• Publication Date: 2020-04-15
• Subjects: neuromuscular junction -- NMJ-morph -- macro -- ImageJ -- Fiji; Science -- Periodicals; 500
• DOI: 10.1098/rsos.200128
• Languages: English
• ISSNs: 2054-5703

Abstract

Abstract : Large-scale data analysis of synaptic morphology is becoming increasingly important to the field of neurobiological research (e.g. 'connectomics'). In particular, a detailed knowledge of neuromuscular junction (NMJ) morphology has proven to be important for understanding the form and function of synapses in both health and disease. The recent introduction of a standardized approach to the morphometric analysis of the NMJ—'NMJ-morph'—has provided the first common software platform with which to analyse and integrate NMJ data from different research laboratories. Here, we describe the design and development of a novel macro—'automated NMJ-morph' or 'aNMJ-morph'—to update and streamline the original NMJ-morph methodology. ImageJ macro language was used to encode the complete NMJ-morph workflow into seven navigation windows that generate robust data for 19 individual pre-/post-synaptic variables. The aNMJ-morph scripting was first validated against reference data generated by the parent workflow to confirm data reproducibility. aNMJ-morph was then compared with the parent workflow in large-scale data analysis of original NMJ images (240 NMJs) by multiple independent investigators. aNMJ-morph conferred a fourfold increase in data acquisition rate compared with the parent workflow, with average analysis times reduced to approximately 1 min per NMJ. Strong concordance was demonstrated between the two approaches for all 19 morphological variables, confirming the robust nature of aNMJ-morph. aNMJ-morph is a freely available and easy-to-use macro for the rapid and robust analysis of NMJ morphology and offers significant improvements in data acquisition and learning curve compared to the original NMJ-morph workflow.