Figure 3
Ependymal localization of A 2B : evidence from immunocytochemistry and X-gal staining. (A) Cytoplasmic A2B-immunoreactivity was evident in ependymal cells (see inset) of wild type mice, although non-specific nuclear labeling was also evident throughout the brain and confounds interpretation of ependymal immunoreactivity. (B) No labeling of ependymal cells was observed using an antibody to A2A receptors in wild type mice, although strong immunoreactivity was evident in the striatum and in a scattered distribution along the SVZ. (C) DIC image from an A2B-/-/β-gal reporter gene knock-in mouse showing darkening of cells due to X-gal precipitate in regions surrounding the ependymal layer (e). Strong X-gal labeling was observed in the lateral septal nucleus (ls), while scattered labeling was observed in the striatum (st) and cortex (ctx) but not in the corpus callosum (cc). The septum mechanically separated from the corpus callosum during the staining procedure, thus obliterating the dorso-medial boundary of the lateral ventricle (lv) in this slice. (Bar = 500 μm). (D-F) Immunocytochemistry from an A2B-/-/β-gal reporter gene knock-in mouse demonstrating that β-galactosidase (D; green, Bar = 25 μm) and S100β (E, red) are co-localized in ependymal cells (F). Nuclei are stained with DAPI (blue). (G) Corresponding DIC image with darkening of the ependyma due to X-gal precipitate.