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. Author manuscript; available in PMC: 2009 Nov 26.
Published in final edited form as: Neuron. 2008 Nov 26;60(4):555–569. doi: 10.1016/j.neuron.2008.09.011

Figure 6. Time-lapse microscopy reveals a brief window for myelination in the maturation of oligodendrocytes.

Figure 6

(A) Time-lapse of a developing OL expressing membrane-targeted EGFP-F as it initiated multiple myelin segments (example indicated by arrowhead) over a brief period (6h–18h), beginning during its sixth day in vitro (6 DIV) with RGC reaggregates in ND-G containing DAPT (1 μM). Following this period, no new clear, stable segments were established (18h–30h). Scale bar = 50 μm.

(B) Dual-color time-lapse of a developing OL expressing EGFP-F (green) under the control of the CMV promoter and mCherry-F (red) under the control of the MBP promoter (MBPp), beginning on the third day (3 d) of coculture in ND-G with DAPT. Multiple new myelin segments were initiated as mCherry-F expression increased, but days 5–6 were marked only by extension (arrowhead), retraction (arrow), and stabilization of existing segments, without formation of new ones. Scale bar = 50 μm.

(C) Dual-transfected OPCs were evaluated for the initiation of new myelin segments and their expression of mCherry-F over 3–4 days. Mature OLs that began with near-maximal expression of mCherry-F rarely initiated new myelin segments, whereas newly differentiated OLs that clearly increased mCherry-F intensity frequently myelinated (% of cells forming at least one new clear myelin segment ± standard error of the proportion, *p<0.01, z-test for independent proportions, n = 52 cells that began with low or sub-maximal mCherry-F expression and 47 cells that began with near-maximal mCherry-F expression).