Figure 5. Oligodendroglial lineage analysis during development.

Postnatal day four through ten C57BL/6 mice were sacrificed, brains extracted, and CNS tissue prepared for flow cytometry (n≥3/time point). Cells were stained with an antibody panel consisting of anti-A2B5, -NG2, -GALC, and -CD45 and analyzed by flow cytometry to characterize changes in oligodendroglial populations. (A) To examine CNS resident cells, single cells were gated, followed by live cells, followed by CD45⁻ gate. The GALC⁺ mature oligodendrocyte population was defined, and the A2B5⁺ and NG2⁺ oligodendrocyte progenitor cell (OPC) populations were defined from the GALC⁻ gate. (b) Representative oligodendroglial analysis from postnatal day 7 and 10 mice. Upper histograms illustrate GALC⁺ mature oligodendrocyte staining. Lower plots indicate A2B5⁺NG2⁻ early OPC and A2B5⁺NG2⁺ intermediate OPC populations. (C) Total cell yields from brains of individual mice, day 4 vs. day 10, P = 0.0013. (D) A2B5⁺NG2⁻GALC⁻ early OPCs (circles), A2B5⁺NG2⁺GALC⁻ intermediate OPCs (squares), and A2B5⁻NG2⁻GALC⁺ mature oligodendrocytes (triangles) are depicted. Error bars represent SD of mice per time point. Early OPCs: day 4 vs. 7, P≤0.001; day 7 vs. 10, P≤0.01. Intermediate OPCs: day 7 vs. 8, P≤0.001; day 8 vs. 10, P≤0.001. Mature oligodendrocytes: day 8 vs. 10, P≤0.05. Data are representative of three independent experiments.