Abstract
Automatic quantification of image parameters is a powerful and necessary tool to explore and analyze crucial cell biological processes. This article describes two ImageJ/Fiji automated macros to approach the analysis of synaptic autophagy and exosome release from 2D confocal images. Emerging studies point out that exosome biogenesis and autophagy share molecular and organelle components. Indeed, the crosstalk between these two processes may be relevant for brain physiology, neuronal development, and the onset/progression of neurodegenerative disorders. In this context, we describe here the macros “Autophagoquant” and “Exoquant” to assess the quantification of autophagosomes and exosomes at the neuronal presynapse of the Neuromuscular Junction (NMJ) in Drosophila melanogaster using confocal microscopy images. The Drosophila NMJ is a valuable model for the study of synapse biology, autophagy, and exosome release. By use of Autophagoquant and Exoquant, researchers can have an unbiased, standardized, and rapid tool to analyze autophagy and exosomal release in Drosophila NMJ.Code available at: https://github.com/IreneSaMi/Exoquant-Autophagoquant
Highlights
The brain is one of the most complex and dynamic organs in the body
We described and report two different macros built with FIJI that serves as a powerful toolbox to quantify autophagosomes and exosomes at the Drosophila Neuromuscular Junction (NMJ)
We describe two macros, Exoquant and Autophagoquant, which improves the standardization in quantifying exosome release and autophagy processes at the synapse
Summary
The brain is one of the most complex and dynamic organs in the body. The most common brain cells are neurons and glia. Different stimuli such as neuronal activity, glucose, calcium, etc., can affect protein expression, localization, and even function This modulation can result in memory consolidation and synaptic plasticity (Rosenberg et al, 2014), synaptogenesis (Kriegstein and Schmitz, 2003), or differentiation (Nazir et al, 2018). The second automatic method quantifies the number, size, and fluorescence intensity of protein clusters/Atg positive dots (representing autophagosomes) inside the NMJ (Figures 1C–C””). This method allows the user to discriminate protein clusters by size, using fluorescence microscope images. This study is the first that completely automatized the task without the researchers decision-making intervention, making it highly reproducible over time and providing new opportunities to compare the data in an objective/unbiased and reproducible way
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
