Abstract
Müller glia (MG) are a relatively quiescent radial glial cell population capable of dedifferentiating to regenerate cells in the zebrafish retina that are lost due to damage. Here, we provide a protocol to both quantify MG cell dedifferentiation behavior during a regenerative response and isolate MG cells by fluorescence activated cell sorting (FACS). First, the retina is exposed to high-intensity light to induce retinal damage and either processed for immunohistochemistry or live MG cells are isolated by FACS that can be used for subsequent genomic or transcriptomic analyses. This method allows us to correlate MG cell behavior observed in situ with their transcriptomic profile at different stages during the regenerative response.
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Acknowledgments
We would like to thank the Faculty of Medicine Flow Cytometry Facility and the Department of Cell & Systems Biology Imaging Facility at the University of Toronto.
Funding: This work was supported by a Vision Science Research Program Scholarship (J.S.), and a OIRM/CFREF-Medicine by Design New Ideas Grant (V.T.) from the University of Toronto.
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Stulberg, J., Tropepe, V. (2022). In Situ Quantification and Isolation of Müller Glial Cells by Fluorescence-Activated Cell Sorting from the Regenerating Larval Zebrafish Retina. In: Kannan, N., Beer, P. (eds) Stem Cell Assays. Methods in Molecular Biology, vol 2429. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1979-7_22
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DOI: https://doi.org/10.1007/978-1-0716-1979-7_22
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Publisher Name: Humana, New York, NY
Print ISBN: 978-1-0716-1978-0
Online ISBN: 978-1-0716-1979-7
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