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bioRxiv · 10.1101/2022.10.14.512256

Versatile and automated workflow for the analysis of oligodendroglial calcium signals in preclinical mouse models of myelin repair

Maas, D. A.·Manot-Saillet, B.·Bun, P.·Habermacher, C.·Poilbout, C.·Rusconi, F.·Angulo, M. C.
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Abstract

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.

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Maas, D. A., Manot-Saillet, B., Bun, P., Habermacher, C., Poilbout, C., Rusconi, F., Angulo, M. C.. 2022-10-18. Versatile and automated workflow for the analysis of oligodendroglial calcium signals in preclinical mouse models of myelin repair. https://doi.org/10.1101/2022.10.14.512256

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