Abstract
LCM-seq is a powerful tool for gene expression analysis from individual or groups of cells that can be spatially isolated. Within the visual system, retinal ganglion cells (RGCs), the cells that connect the eye to the brain through the optic nerve, reside in the retinal ganglion cell layer of the retina. This well-defined location provides a unique opportunity to harvest RNA by laser capture microdissection (LCM) from a highly enriched cell population. Using this method, it is possible to explore transcriptome-wide changes in gene expression following optic nerve injury. In the zebrafish model, this method can be used to identify molecular events driving successful optic nerve regeneration in contrast to mammals that fail to regenerate axons in the central nervous system. Here we provide a method for LCM from the different retinal layers of zebrafish following optic nerve injury and during the process of optic nerve regeneration. Purified RNA from this protocol is sufficient for RNA-seq or other downstream analysis.
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Acknowledgments
Funding for this project was provided to MBV by a pilot grant from the Medical College of Wisconsin Research Affairs Committee.
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© 2023 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature
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Speer, W., Veldman, M.B. (2023). LCM-Seq for Retinal Cell Layer-Specific Responses During Optic Nerve Regeneration. In: Udvadia, A.J., Antczak, J.B. (eds) Axon Regeneration. Methods in Molecular Biology, vol 2636. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3012-9_17
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DOI: https://doi.org/10.1007/978-1-0716-3012-9_17
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Publisher Name: Humana, New York, NY
Print ISBN: 978-1-0716-3011-2
Online ISBN: 978-1-0716-3012-9
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