Characterizing oligodendroglial population changes through cuprizone-induced demyelination.

<p>C57BL/6 mice were fed 0.2% cuprizone chow for five weeks and returned to a normal diet for a sixth week. A cohort of mice was sacrificed weekly throughout the disease course, brains were extracted, and CNS processed for flow cytometry (n = 5). Naïve mice were concurrently assayed at each time point for standardization (n = 3). (<b>A</b>) Cells were first gated by forward and side scatter then single cells were gated. Live cells were next gated, and hematopoietic cells excluded by gating on CD45^-/low cells. (<b>B</b>) From the CNS resident cells, oligodendroglial populations were defined: A2B5⁺PDGFRα⁺ early OPCs, PDGFRα⁺O4⁺ intermediate OPCs, O4⁺O1⁺ pre-myelinating oligodendrocytes, and O1⁺GALC⁺ mature oligodendrocytes. Mature oligodendrocytes (<b>C</b>), pre-myelinating oligodendrocytes (<b>D</b>), early OPCs (<b>E</b>), and late OPCs (<b>F</b>) were compared to naïve mice at each time point. CD45^high peripheral immune cells (<b>G</b>) and CD45^low microglia (<b>H</b>) were also quantified. Error bars indicate SD of mice per time point. *<i>P</i>≤0.05; **<i>P</i>≤0.01; ***<i>P</i>≤0.001 for post-hoc analysis. Data are representative of two independent experiments.