Research Paper Volume 8, Issue 7 pp 1540—1570

Figure 4. Newborn CCNA2^fl/fl, Nestin-cre animals exhibit a developmental delay. (A) P0 animals were pulsed with BrdU for 30 minutes before euthanasia and tissue stained for BrdU and Pax6. (B) Quantifications of BrdU, Ki67, and pH3 were significantly elevated in CCNA2^fl/fl, Nestin-cre mice. The y-axis is the percentage of cells positive for each marker in the VZ/SVZ. Unpaired t-test, * = p < 0.05. (C) P0 brains were sectioned for unbiased stereology, stained for NeuN, and counterstained with hematoxylin. NeuN-positive cells were quantified in the combined VZ/SVZ and intermediate zone (IZ). Experimental conditions are indicated above, molecular marker is color-coded on the left. VZ/SVZ, intermediate zone (IZ), and CP are indicated. (D) Optical fractionator measurements of NeuN-positive cells in the VZ/SVZ/IZ show increased neuronal output in CCNA2^fl/fl, Nestin-cre animals compared to controls, indicative of a developmental delay. The y-axis is the number of NeuN-positive cells in the VZ/SVZ/IZ per brain. Unpaired t-test, * = p < 0.05, n = 3 brains per condition. (E) Measurement of the number of sections sampled and (F) measurement of the number of sampling sites. There was no significant difference in either metric, precluding the possibility that the increased NeuN was due to a difference in brain size. For (E), the y-axis is the number of sections sampled per brain. For (F), the y-axis is the number of sampling sites per brain. Unpaired t-test, n.s. = not significant, p > 0.05, n = 3 brains per condition. Error bars for all graphs represent s.e.m.