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. 2004 Feb 12;101(9):3196–3201. doi: 10.1073/pnas.0308600100

Fig. 2.

Fig. 2.

Not only a subpopulation of the mitotic cells at the apical side of the neuroepithelium, but also mitotic cells in the basal portion of the VZ and in the SVZ, show Tis21-driven GFP expression. (a-h) 4′,6-Diamidino-2-phenylindole staining (white; a, c, e, and g), GFP fluorescence (Green; b-d and f-h), and βIII-tubulin immunofluorescence (red; d and h) of the E10.5 telencephalon (a-d), E10.5 hindbrain (e-g), and E9.5 hindbrain at lower magnification (h). Solid white lines, basal lamina; dashed white lines, apical surface; arrows, GFP-expressing NE cells in interphase; open white arrowheads, mitotic NE cells lacking GFP. (a-g) Scale bar in top right corner of d, 20 μm. (a-d) Note the mitotic cell (open arrows with asterisk) in the basal portion of the VZ, which expresses GFP (b-d) but not βIII-tubulin (d), in contrast to the young neurons (solid white arrowheads; NL, neuron-containing layer), which contain both GFP (b-d) and βIII-tubulin (d). (e-g) At the apical surface of the VZ, mitotic NE cells lacking GFP are observed (open white arrowheads), which may coexist with mitotic NE cells expressing GFP (open arrows). (h) A newborn neuron (solid white arrowhead) in the VZ. (i) Low-power overview of the E13.5 telencephalon showing mitotic cells, identified by phosphohistone H3 immunostaining (red), at the apical surface (dashed white line) of the VZ and in the SVZ but not in the overlying neuron-containing layer (NL; solid white line, basal lamina). Mitotic cells in the VZ and SVZ showing GFP fluorescence (green) upon individual inspection at higher magnification (not shown) are indicated by open and filled white arrowheads, respectively; note that in the merged image shown, the phosphohistone H3 immunostaining (red) largely obscures the signal for GFP fluorescence, except for mitotic cells with high GFP levels (yellow). (j and k) Quantitation of mitotic GFP-positive cells in the basal portion of the VZ (plus, when present, the SVZ) of the telencephalon at E10.5 and E13.5 (j) and hindbrain at E10.5 and E12.5 (k), expressed as percentage of total mitotic cells in the basal portion of the VZ (plus, when present, the SVZ) (4-140 cells counted per embryo). (l and m) Quantitation of mitotic GFP-positive cells at the apical side of the neuroepithelium (A, diamonds) and in the basal portion of the VZ (plus, when present, the SVZ) (B, triangles), each expressed as percentage of total mitotic cells at the apical side of the neuroepithelium (130-900 cells counted per embryo), as well as the sum of both (A+B, squares), in the telencephalon (l) and hindbrain (m) at various developmental stages. (j-m) Data are the mean of two embryos (except for E9.5, which is a single embryo); bars indicate the variation of the individual values from the mean and are sometimes included within the symbol.