PT  - JOURNAL ARTICLE
AU  - Maas, Dorien A.
AU  - Manot-Saillet, Blandine
AU  - Bun, Philippe
AU  - Habermacher, Chloé
AU  - Poilbout, Corinne
AU  - Rusconi, Filippo
AU  - Angulo, Maria Cecilia
TI  - Versatile and automated workflow for the analysis of oligodendroglial calcium signals in preclinical mouse models of myelin repair
AID  - 10.1101/2022.10.14.512256
DP  - 2022 Jan 01
TA  - bioRxiv
PG  - 2022.10.14.512256
4099  - http://biorxiv.org/content/early/2022/10/18/2022.10.14.512256.short
4100  - http://biorxiv.org/content/early/2022/10/18/2022.10.14.512256.full
AB  - Intracellular Ca2+ signals of oligodendroglia, the myelin-forming cells of the central nervous system, regulate vital cellular processes including myelination. However, studies on oligodendroglia Ca2+ signal dynamics are still scarce, especially during myelin repair, and there are no software solutions to properly analyze the unique Ca2+ signal characteristics in these cells. Here, we provide a comprehensive experimental and analytical workflow to acquire and analyze Ca2+ imaging data of oligodendroglia at the population and single-cell levels in preclinical mouse models of myelin repair. We report diverse ex vivo and in vivo experimental protocols to obtain reproducible Ca2+ imaging data from oligodendroglia in demyelinated lesions. Importantly, we provide an analytical pipeline containing two free, open source and cross-platform software programs, Occam and post-prOccam, that enable the fully automated analysis of one- and two-photon Ca2+ imaging datasets from oligodendroglia obtained by either ex vivo or in vivo Ca2+ imaging techniques. This versatile and accessible experimental and analytical framework, which revealed significant but uncorrelated spontaneous Ca2+ activity in oligodendroglia inside demyelinated lesions, should facilitate the elucidation of Ca2+-mediated mechanisms underlying remyelination and therefore help to accelerate the development of therapeutic strategies for the many myelin-related disorders, such as multiple sclerosis.Competing Interest StatementThe authors have declared no competing interest.