Figure 3

Identification of neural cell type containing TNFα protein. This photographic collage shows the result of co-localization studies identifying the neural cell type containing TNFα protein in the brain of the transgenic animal. All images were taken of TNFα-transgenic rat brain that had not been subjected to ischemic injury. Coronal brain sections were incubated with primary antibodies identifying mouse or rat TNFα protein and neurons. All images were made by fluorescence microscopy at 40× magnification. In Panel a, cortical neurons in TNFα-Tg rat brain are immuno-labeled with the anti-neuronal antibody NeuN tagged with a green fluorophore. In Panel b, co-localization images are merged to show that several NeuN-labeled neurons (green) express TNFα protein immuno-labeled with anti-mouse/rat TNFα antibody tagged with a red fluorophore. In Panel c, co-localization images are merged to show NeuN-labeled neurons (green), TNFα protein (red), and cellular nuclei labeled with Hoechst stain (blue). A representative neuron with cytoplasmic inclusions labeled to show expression of TNFα protein is identified in each Panel (arrow). In Panel c, nuclei of several non-neuronal cells that do not express TNFα protein are shown. Scale bar = 50 μm