Fig. 6

Effect of IL1RAPL 1B and IL1RAPL 1B-P455H expression on the VAMP2-EGFP punctum formation in axon terminals of olfactory sensory neurons. (A) Omp promoter-driven expression vectors for VAMP2-EGFP (Pomp-VG, top), for IL1RAPL 1B and VAMP2-EGFP (Pomp-VG-Il1rapl1b, middle) and for IL1RAPL 1B-P455H and VAMP2-EGFP (Pomp-VG-P455H, bottom) in olfactory sensory neurons. Black boxes, the omp promoter; crosshatched boxes, the 3′ downstream sequence of the omp gene; hatched boxes, SV40 polyadenylation signal sequence; lines, pBluescript II SK+. (B-D) Representative VAMP2-EGFP signals in olfactory neuron axon terminals in Pomp-VG (B)-, Pomp-VG-Il1rapl1b (C)-or Pomp-VG-P455H (D)-injected embryos at 36 hpf (left), 60 hpf (middle) and 84 hpf (right) are shown in the upper panels. The threshold images of VAMP2-EGFP signals are on the bottom. Arrowheads point axonal shafts. Bar = 5 μm. (E) The area of VAMP2-EGFP puncta in axon terminals of olfactory sensory neurons in Pomp-VG (open bars)-, Pomp-VG-Il1rapl1b (filled bars)-and Pomp-VG-P455H (stippled bars)-injected embryos at 36 hpf, 60 hpf and 84 hpf. Pomp-VG, n = 33, 41 and 40 for 25, 29 and 27 embryos at 36 hpf, 60 hpf and 84 hpf, respectively; Pomp-VG-Il1rapl1b, n = 41, 39 and 49 for 25, 27 and 26 embryos at 36 hpf, 60 hpf and 84 hpf, respectively; Pomp-VG-P455H, n = 41, 43 and 44 for 26, 27 and 27 embryos at 36 hpf, 60 hpf and 84 hpf, respectively. Two-way ANOVA, expression vector effect, F = 27.9, p < 0.001. All values represent mean ± SEM. ** and ***, p < 0.01 and< 0.001, respectively.