Figure 5
An Mbp promoter induced transgene expression into oligodendrocytes specifically in the mouse cerebrum. (A) Schematic of pPB-Mbp-EGFP and pCAG-PBase plasmid. pPB-Mbp-EGFP drives EGFP expression under an Mbp promoter. (B) Experimental procedure. pPB-Mbp-EGFP and pCAG-PBase were co-electroporated into the mouse cerebrum at E15.5, and coronal sections were prepared at P30. (C) Coronal sections of the electroporated cerebrum. Most EGFP-positive cells were accumulated in the white matter. (D–G) Immunohistochemistry for NeuN (D), Sox9 and Sox10 (E), and in situ hybridization for Plp1 (F). Most EGFP-positive cells were positive for Plp1 and negative for NeuN and Sox9 (open arrowheads). (G) The percentages of EGFP-positive cells which were also NeuN-positive (neuron), Sox9-positive/Sox10-negative (astrocyte) or Plp1-positive (oligodendrocyte). This Mbp promoter drove gene expression in oligodendrocytes selectively when combined with the piggyBac system and in utero electroporation. Unpaired Student’s t-test, *P < 0.00005. Error bars represent mean ± SD. The statistical analyses were performed using Microsoft Excel ver. 16.43. Scale bars: 200 μm (C), 30 μm (D-F). TR, terminal region; WM, white matter. Numbers indicate layers in the cerebral cortex.
