Morphology and cellularity changes in wild type and BNIP3 knockout brains.

Whole brains from 8-week old wild type and BNIP3-/- mice were fixed by cardiac perfusion and overnight immersion in paraformaldehyde (n = 3 pairs of littermates from different heterozygous crosses). Brains were embedded in paraffin, sectioned and stained with hematoxylin and eosin as described in Materials & Methods. Images were captured at 10x and 40x magnification and cell counting was performed using Image Pro Plus 5.0 software. These representative images depict the morphology of the cerebellum, hippocampus, cortex and inferior colliculus, with cell counts for the 40x images indicating increased cellularity in BNIP3-/- brains. E18.5 embryos from B) wild type and C) BNIP3-/- mice were fixed as described in the Material and Methods section. The paraffin embedded brains were sliced and placed on slides and stained for DNA with DAPI and antibodies against Ki67 and leaved caspase 3. This images represent three independent experiments.