Figure 5.
Proteasome Inhibition Changes ASPD Distribution from Axon to Dendrites
(A) Upper panels: primary rat hippocampal neuronal cultures with AAV-APPswe transduction were quadruple stained at 30 DIV with the anti-ASPD rpASD1 antibody, anti-NFH antibody, anti-MAP2 antibody, and DAPI (see Transparent Methods). Lower panels: primary rat hippocampal neuronal cultures with AAV-APPswe transduction were treated with 75 nM MG132 for 24 h and quadruple stained at 22 DIV as in the upper panels. A representative image, along with the corresponding single red, green, or magenta image, is shown with 50-μm scale bars. Orange arrows mark the axon, whereas white double-lined arrows mark dendrites. ASPD were co-localized with NFH (orange arrows) in the upper panels, but co-localized with MAP2 (white double-lined arrows) in the lower panels.
(B) APPswe-transduced primary rat hippocampal neuronal cultures were treated with 75 nM MG132 for 24 h. The cultures were triple stained with antibodies against MAP2, Bassoon, and ASPD (rpASD1) on the left side. Representative images are shown. Scale bar, 1 μm.
(C) Spatiotemporal changes in the ASPD distribution observed in Figures 3B and 4A are schematically shown in red.