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. 2009 Jun 24;29(25):7966–7977. doi: 10.1523/JNEUROSCI.1054-09.2009

Figure 4.

Figure 4.

GABA-mediated depolarization regulates CREB phosphorylation, differentiation, and morphological maturation of new dentate granule cells during the first 2 weeks after their birth. a, Analysis of CREB phosphorylation and DCX expression after shRNA-mediated knockdown of NKCC1. Confocal images of control shRNA-transduced cells (shCTR) (green) and shNKCC1-transduced cells (green) at 14 dpi. Bottom, Magnifications of indicated boxes. A large fraction of shNKCC1-transduced DCX+ (blue) immature neurons is negative for pCREB (red) (white arrowhead). Moreover, many shNKCC1-transduced cells do not express DCX (yellow arrowhead). pCREB+-transduced immature neurons (red arrowheads). Scale bars, 25 μm. b, The percentage of pCREB-positive immature neurons (identified by DCX expression) among the shNKCC1-transduced immature neurons is significantly lowered (***p < 0.001). c, Expression of DCX is significantly reduced among shNKCC1-transduced cells (***p < 0.001). d, Analysis of NeuroD expression (red) in newborn cells after transduction with shCTR (green) or shNKCC1 (green) at 14 dpi. Scale bar, 25 μm. A large percentage of shNKCC1-transduced cells do not express NeuroD (white arrowheads). Red arrowheads indicate transduced cells, which express NeuroD. e, Quantification of NeuroD expression in transduced cells. Expression of NeuroD is significantly reduced among the shNKCC1-transduced cells (**p < 0.01). f, The percentage of shNKCC1-transduced immature neurons (DCX+), which extend an apical dendrite through the granule cell layer toward the molecular layer, is significantly reduced (**p < 0.01).