Figure 2.

Radial Glial Cells Generate Ependymal Cells and Adult Neural Stem Cells (Type B1 Astrocytes)

(A) Experimental schematic for (B)–(D). The H2B-GFP-expressing plasmid was electroporated in utero at E14.5 and analyzed on V-SVZ whole-mount (WM) at P15. CC, corpus callosum; Cx, cortex; LV, lateral ventricle; R, rostral; D, dorsal.

(B and D) P15 V-SVZ whole-mounts were double-immunostained with FoxJ1 (red) and Sox9 (blue) antibodies (B) or FOP (white) and GFAP (red) antibodies (D). GFP⁺FoxJ1⁺Sox9⁺ ependymal cells are indicated by arrows, and GFP⁺FoxJ1⁻Sox9⁺ astrocytes are outlined in white (B). GFP⁺GFAP⁻ ependymal cells with multiple FOP⁺ dots are indicated by arrows, and a GFP⁺GFAP⁺ astrocyte with a FOP⁺ centrosome is indicated by an arrowhead (D).

(C) Mean percentage of astrocytes (Sox9⁺FoxJ1⁻), ependymal cells (Sox9⁺FoxJ1⁺), and others (Sox9⁻FoxJ1⁻) among H2B-GFP⁺ electroporated cells. Analyses were done on n = 3 animals; a total of 441 cells were counted. Error bars represent the SEM. The p values were determined with a two-proportion Z test; ^∗∗∗p ≤ 0.001, ^∗∗p ≤ 0.01.

(E) Experimental schematic for (F) and (G). Nucbow plasmids (^PBCAG-Nucbow along with the PiggyBac transposase and the self-excising Cre recombinase) were electroporated in utero at E14.5 and received EdU (through drinking water) for 14 days starting at P21.

(F and G) Coronal sections of the olfactory bulb (OB) were prepared 1 week after the last day of EdU administration. (G) is a high-magnification image of (F) to show that some Nucbow⁺ interneurons in the OB are EdU⁺.

The scale bars represent 40 μm (B), 15 μm (C), 520 μm (F), and 180 μm (G).