Circadian Clock Genes Are Essential for Normal Adult Neurogenesis, Differentiation, and Fate Determination
Fig 5
Neuronal commitment is diminished in Bmal1-/- DG neurospheres.
In WT DG neurospheres the sequence of cell types during differentiation in B27 medium parallels events during in situ neurogenesis. A: Immature neurons expressing BetaIII-tubulin (green) at day 7 and lacking GFAP co-localization (red). B: Mature neurons expressing NeuN (green) at day 14 and lacking GFAP (red). In contrast, Bmal1-/- neurospheres displayed reduced neuronal differentiation and increased astrocyte proliferation. C: Lack of BetaIII-tubulin expression (green) shown with GFAP (red) in a Bmal1-/- neurosphere at day 7. D: Lack of NeuN (green) shown with GFAP (red) at day 14. All nuclei were stained with Hoechst (blue). Scale bar = 50 μm. E: Percentage of positive cells for DCX (neuroblasts), BetaIII-tubulin, and NeuN at days 4, 7, and 14 after differentiation in B27 medium, respectively.