Fig. 3

Focal brain lesion in an alpaca (Vicugna pacos): Fluorescence labeling to illustrate lesions at the border of the damaged tissue (a–c) as well as in in the core of affected neocortical brain regions (d, e). (a–c): Double staining of microglia/macrophages (Iba/STL) and vascular endothelial cells (STL) combined with the detection of neuronal nuclei (NeuN). The dashed line (a, b) separates the altered tissue (upper left part) from nonaffected brain parenchyma. In the upper left part (a), Iba-immunolabeling (Cy3, red fluorescence) reveals ameboid microglia/macrophages, simultaneously visualized in (b) by STL-binding sites (Cy5-staining, color-coded in blue). Additionally, the lectin-histochemical Cy5-staining (color-coded in blue) detects some endothelial cells. The merged staining patterns of Iba, STL and NeuN in (c) clearly show the neuronal loss in the gliotic tissue. (d, e): Simultaneous detection of the lesion markers collagen IV as component of the vascular basement membrane and neuronal NF-L counterstained by STL detecting vascular endothelial cells. In d, strong Cy2-immunostaining of collagen IV (Coll, Cy2, green) indicates lesioned tissue, whereas in (e) the merged staining patterns of Coll (Cy2, green) and STL (Cy5, color-coded in blue) results in the turquoise appearance of vessels. These are located in close vicinity to pyramidal cells displaying enhanced NF-L-immunoreactivity (Cy3, red). Scale bars: c (also valid for a and b): 100 µm, d (also valid for e): 150 µm