Figure 6.
Brain endothelial cells express N-methyl--aspartate (NMDA) receptor subunits. (A) Purified primary adult mouse brain endothelial cultures expressed mRNA for housekeeping gene, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), and NR1. C denotes cortex; E denotes endothelial culture; B denotes blank control. (B) Western blot of protein from cortex (C) and endothelial cultures (E) produced a band for NR1 at 116 kDa. (C–E) Immunofluorescence shows colocalization of NR1 (C, green) and the endothelial cell marker, isolectin B4 (D, red), in cultured endothelial cells. Nuclei were stained with ToPro-3. Scale bars for (C–E) are 100 μm. (F–H) Coimmunofluorescence of NR2C (F, red) and p-glycoprotein (G, green) in cortical arteries near the pial surface. (I–K) Immunofluorescence for NR2C (I, red) and astrocyte glial fibrillary acidic protein (GFAP; J, green) does not overlap (K) in cortical arteries near the pial surface. Scale bars for (F–K) are 50 μm.