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. 2016 Apr 5;12(4):e1005551. doi: 10.1371/journal.ppat.1005551

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Fig 4 — Cell-type associations of PrPSc in cortex, substantia nigra and hypothalamus after striatal microinjection of scrapie strains 22L (A-D), RML (E-H) or ME7 (I-L). PrPSc was detected by IHC with monoclonal antibody D13 (brown color), and some slides were also dual-stained with a second antibody to detect neurons (NeuN) or astroglia (GFAP) as described in methods. (A) At 40 dpi 22L PrPSc is associated with a stellate GFAP-positive astrocyte (arrow). Nearby larger neurons and vascular endothelial cells are PrPSc-negative. (B) At 40 dpi stellate cells (arrows), likely to be astrocytes, are positive for 22L PrPSc and are intermingled with neurons stained with anti-NeuN (pink). (C) At 40 dpi in substantia nigra 22L PrPSc is not associated with Neu-N-positive neurons (pink) but instead surrounds cells with small blue nuclei, possibly astrocytes. (D) In hypothalamus at 60 dpi single stained 22L PrPSc surrounds small cells with blue nuclei (arrows) and is less often adjacent to round neurons with prominent nucleoli. (E) At 60 dpi RML PrPSc is associated with a GFAP-positive astrocyte (arrow) and also with perivascular astrocyte surrounding a small blood vessel (arrowhead). (F) In cortex at 60 dpi RML PrPSc surrounds cells with small nuclei (arrows) near Neu-N-positive neurons, similar to 22L in panel (B). (G) In substantia nigra at 60 dpi RML PrPSc is associated with cell surface of two Neu-N-positive neurons (arrows). PrPSc is also seen in the neuropil between neurons (arrowhead). (H) At 80 dpi RML PrPSc in hypothalamus, detected by single staining with D13, is associated with neuropil (arrowheads), blood vessel (blue arrow) and probable astroglia with small nuclei (black arrow). (I) At 100 dpi ME7 PrPSc in cortex is deposited in fine and coarse aggregates in neuropil (arrowheads), but is not associated with GFAP-positive astroglia (arrow). (J) At 100 dpi in cortex ME7 PrPSc is associated with NeuN-positive neurons (arrows). Occasional coarse aggregates are also seen in the neuropil (arrowhead). (K) At 60 dpi in substantia nigra ME7 PrPSc surrounds some Neu-N-positive neurons and also is seen as punctate deposits (arrowheads) near neurons and as linear staining suggestive of axons. (L) At 100 dpi in hypothalamus ME7 PrPSc is shown using single staining in a perineuronal distribution (arrows) and also as coarse neuropil aggregates (arrowheads). Scale bar shown in panel A is 50 μm and applies to all panels.