Chapter 2 52 Figure 6. Oligodendrocyte maturation and myelination in co-culture with 4H neurons is normal. A and B indicate that neuron-oligodendrocyte co-cultures show robust generation of oligodendrocytes and myelination. (A) Staining for myelin protein MBP and axonal marker NF200 shows wrapping of oligodendrocyte processes around axons. (B) Staining for MBP and early oligodendrocyte marker OLIG2 shows a robust generation and maturation of oligodendrocytes. (C) Electron microscopy confirms the presence of compact myelin sheets around neuronal processes. (D) Cultures with 4H or control neurons and control oligodendrocytes both show oligodendrocyte maturation and myelination. Quantification showed no changes in oligodendrocyte generation, defined as the percentage of DAPI+ cells that were OLIG2+ (E). Similarly, oligodendrocyte maturation (F; percentage of OLIG2+ cells that developed in MBP+ cells) or the percentage of neurites that is myelinated (G) was not changed in cultures with 4H neurons. (A and D) Scalebar = 25 μm, (B) scalebar = 100 μm, (C) scalebar = 500 nm. (E-G) Open circles represent average per individual patient/control, filled circles represent the mean per genotype. Error bars represent SEM.
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