Document Type
Article
Publication Date
2026
Publication Title
Current Protocols
Keywords
calcium imaging, chill coma, migraine, spreading depolarization, spreading depression
Abstract
Spreading depolarization (SD) is a ubiquitous neural phenomenon implicated in mammalian neurological disorders including cerebral ischemia, brain death, stroke, brain injury, and migraine, as well as in insect chill coma. During SD, a breakdown of ion homeostasis and a slowly spreading wave of neural depolarization leads to processing deficits, such as sensory disturbances and paralysis. Our protocols introduce high-throughput techniques using wide-field fluorescence brain imaging in Drosophila melanogaster larvae and adults to detect and quantify the spatial and temporal dynamics of SD and its initiation points. Two SD initiation techniques, rapid cooling and pharmacological manipulation, are detailed. The larval protocols describe how to elicit, detect, and quantify SD in up to 16 animals simultaneously in either intact or dissected preparations. The adult SD protocols describe the detailed steps to elicit, detect, and quantify SD in either fully intact animals or after microdissection that enables high resolution imaging of the brain. Detection, quantification, and analysis techniques are described in a separate protocol. Our protocols establish the fly as a high-throughput model for studying conserved cellular and circuit-level mechanisms of SD initiation and propagation.
Funding Source
Stocks obtained from the Bloomington Drosophila Stock Center (NIH P40OD018537) were used in this study. This article was published open access thanks to a transformative agreement between Milner Library and Wiley.
Creative Commons License
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License
DOI
10.1002/cpz1.70475
Recommended Citation
Stein, W., & Harris, A. L. (2026). Detection and quantification of spreading depolarization in drosophila melanogaster using high-throughput, wide-field brain imaging. Current Protocols, 6, e70475. https://doi.org/10.1002/cpz1.70475
Comments
First published in Current Protocols (2026): https://doi.org/10.1002/cpz1.70475