Figure 3

Fractin and TubulinΔCsp6 labeling highlight degenerating axon tracts after ethanol-induced apoptosis. (a) Low power image of co-labeling in the striatum using anti-tubulinΔCsp6 (cTub; green) along with anti-neurofilament (NFM; red) reveals that tubulinΔCsp6 localizes to axon tracts 24 hours post-ethanol treatment. (b-c) High power image of co-labeling in the striatum using fractin or TubulinΔCsp6 (green) along with a marker for neurofilaments (red) reveals that both fractin and TubulinΔCsp6 highlight blebbing material within axon tracts after ethanol-induced apoptosis (b,d) and are absent in control (saline-treated) brain (c,e). (f-g) While ethanol-induced apoptosis led to CC3-positive cell bodies, CC3 was not usually seen in axon fibers via conventional staining techniques. Tyramide signal amplification (TSA) was required for visualization of CC3 in axon fibers after ethanol-induced apoptosis (shown are striatal pencil fibers). CC3 with TSA (CC3 + TSA) was only detected in tissue after ethanol-induced apoptosis (f), and CC3 staining was not seen in control tissue (g). Scale in (a) is 200 μm. Scale in (b-g) is 50 μm.