Figure 2.
Adenoviral and lentiviral codelivery to the dorsal subpia can be used mark dorsal radial glia and descendents. A, Brains are selected for analysis when they meet the criteria that the cortical surface is not damaged and the dsRedExpress containing lentivirus only infects cells in the outer cortical layer. Three weeks after codelivery of the lentivirus and a Cre-containing adenovirus to R26R-YFP mice at P1/2, the lentivirus remains confined to subpial astrocytes (red). In contrast, the adenovirus has been retrogradely transported to cells throughout the cortex that had pial attachments at the time of viral delivery, and these cells can be traced long term by examining YFP fluorescence. In the cortex, YFP+ astrocytes can be observed. B, Four days after infection, astrocytes and RC2+ radial glial processes (inset) can be seen in the radial strip from the subpial surface to the subventricular zone. Brains are only considered for analysis when the area on the cortical surface that was in contact with the adenovirus (demarcated by red lines) is <1 mm in diameter. C, Dorsolateral subventricular zone (borders outlined in blue) at 4 d after infection reveals YFP+ cells along the dorsal border. D, Expanded view of boxed region in C reveals colocalization of YFP with the radial glial and astrocytic marker Zebrin II (ZII). E, F, Three weeks after labeling dorsal radial glia with the adenovirus, NG2+ oligodendrocyte precursors (E) and CC1+ oligodendrocytes (F) are observed in the subcortical white matter. Dorsal is up in A–D. The dotted line demarcates ventricle. V, Ventricle; P, pial surface. Scale bar, 20 μm (in all images).