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. 2013 Jun 13;22(21):4253–4266. doi: 10.1093/hmg/ddt276

Figure 9.

Figure 9.

PrP-WT enhances the neurotoxicity of α3M>S mutants. (A) Micrographs show maximum projection of Z-stacks of mushroom body lobes. Two copies of PrP-WT (2×) result in normal mushroom bodies followed by generalized blebbing in older flies, arrowhead) with thinning of both dorsal and medial projections. Co-expression of PrP-WT and –M205S induces abnormal development of the dorsal lobe, arrow) and progressive degeneration of all lobes co-expression of PrP-WT and -M205,212S results in thin and short dorsal and medial projections, arrows) followed by potent degeneration of all lobes after 40 days, arrows) and overall blebbing, arrowhead). (B) Statistical analysis of the mushroom body surface. N = 12 for each group. ***P < 0.001. (C) Locomotor activity of flies co-expressing WT and mutant PrP. Control flies expressing LacZ under the control of BG380-Gal4 show strong climbing ability (black dashed line). Flies expressing 2×WT stop climbing at Day 6 (black lines). Flies expressing PrP-WT + M205S (grey line) stop climbing at Day 4. Flies co-expressing PrP-WT + M205,212S (black dotted line) start with lower climbing ability and stop climbing at Day 5. ***P < 0.001. (D and E) Immunoprecipitation with 15B3. (D) Using 6H4, PrP-WT and -M205S accumulate 15B3-positive conformations at Day 30, but 6H4 does not recognize PrP-M205,212S. (E) 6D11 does not recognize 15B3-positive conformations in PrP-WT flies, but detects them in PrP-M205S at Day 30. PrP-M205,212S immunoreacts with 15B3 at Day 1 and accumulates at higher levels at Day 30. On the left, controls with no 15B3 or no PrP demonstrate the specificity of the assay. (F) 15B3 immunoprecipitation in flies co-expressing WT and mutant PrPs. PrP-2×WT flies accumulate 15B3-positive isoforms at Day 30. PrP-WT + M205S flies show accumulation of 15B3 conformations at Day 1 corresponding to PrP-M205S. In Day 30 flies, both PrP-WT and -M205S are recognized by 15B3. In flies expressing PrP-WT + M205,212S, both PrP-WT and -M205,212S accumulate 15B3-positive isoforms from Day 1.