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. 2009 Apr 1;29(13):4172–4188. doi: 10.1523/JNEUROSCI.4956-08.2009

Figure 3.

Figure 3.

Expression of BDNF in the adult RMS. A, In situ hybridization (ISH) for BDNF and immunostaining for PECAM (green) showing that endothelial cells of blood vessels synthesize BDNF. Note that BDNF mRNA is present in the endothelial cells (arrowheads) but not in other cell types of the adult RMS (asterisks). Scale bar, 5 μm. B, RT-PCR analysis of FACS-purified endothelial cells (Endoth.), astrocytes (Astro), and neuroblasts (Neuro). BDNF is expressed in endothelial cells but not in astrocytes and neuroblasts. C, Immunostaining for BDNF in the adult mouse RMS. Inset shows colocalization of BDNF (green) and GFAP (red). Scale bar, 10 μm. D, Immunostaining for BDNF (green) and GFAP (red) in the adult mouse RMS after osmotic minipump infusion of TrkB–Fc or IgG–Fc, as a control. BDNF immunostaining is drastically reduced after 7 d of TrkB–Fc infusion. Scale bar, 10 μm. E, Coculture of astrocytes with BDNF–GFP-transfected endothelial cells. Note the strong GFP signal in the endothelial cells and punctate staining (arrows) of astrocyte. Bright-field (BF) images are also shown. Bleaching area is indicated by the red square. After 8 min of recovery period, the punctate staining reappears at the surface of astrocyte. Scale bar, 10 μm.