Figure 5

HDAC3 expression in normal brain and cell-specific expression in the cerebellum. (A) Immunohistochemical staining of sections from 10.5-month-old non-transgenic control mouse. Fixed tissue sections were probed with an antibody to HDAC3 conjugated to Alexa 488. Multiple mouse brain regions were captured. Top panel L-R: hippocampus CA3 region; dentate gyrus (DG) and subgranular zone (SGZ); and level IV-V motor cortex with pyramidal neurons visible; Bottom panel L-R: granular cell layer (GCL) and purkinje cell layer (PCL) of the cerebellum; pons (Po); caudate putamen (CPu) and subventricular zone (SVZ). Scale bar is 40 μM. (B) Immunohistochemical staining of sections from the cerebellum of 12-month-old non-transgenic control mouse. Fixed tissue sections were probed with an antibody to HDAC3 conjugated to Alexa 488 (green) and antibodies to various cell-specific markers conjugated to Alexa 555 (red) and counterstained with DAPI (blue). NeuN marks neuronal nuclei and colabeling with HDAC3 is particularly evident in the upper nuclear layer (arrows), also called the purkinje cell layer (PCL). Calbindin marks purkinje cells with HDAC3 colocalization detected both in the cell bodies and processes (arrows). GFAP marks neuroglia. Colocalization with HDAC3 is detected in the processes of Bergman glia (radial glial cells; arrows) in the molecular layer (ML) as well as in astrocytes in the granular cell layer (GCL; arrowhead). High-resolution image with S100 calcium-binding protein beta (S100β) staining specifically of Bergmann glia cells, with discrete HDAC3 colocalization in cell bodies and along processes (arrows). Scale bar represents 10 μm.