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Figure 2. Acute Inhibition of Translation Disrupts Axonal Branching Dynamics In Vivo(A) Live imaging experiment on branching dynamics of somaless RGC axons in the tectum in vivo. Electroporated eye was removed to eliminate somatic contribution.(B–D) Axonal branching in control condition (B) and after incubation in translation inhibitors cycloheximide (C; CHX) and anisomycin (D; ANI). A merged overlay of three time points (0′, 5', and 10′ in blue, red, and green, respectively) is shown for each condition (far right). More protrusions were added than removed in control condition (filopodia: t11 = 3.8, p = 0.003; branches: t11 = 4.6, p = 0.0008) (B′ and B″). No significant differences were observed in the number of protrusions that were added and removed in CHX condition (filopodia: t18 = 0.2, p = 0.82; branches: t18 = 1.1, p = 0.29) (C′ and C″). No significant differences were observed in the number of protrusions that were added and removed in ANI condition (filopodia: t21 = 0.5, p = 0.66; branches: t21 = 1.4, p = 0.18) (D′ and D″).(E and F) The dynamics of filopodia (E; addition: F2,50 = 18.7, p < 0.0001; removal: F2,50 = 13.0, p < 0.0001) and branches (F; addition: F2,50 = 20.2, p < 0.0001; removal: F2,50 = 9.5, p = 0.0003) were inhibited by CHX or ANI treatment.Error bars represent SEM. ∗∗p < 0.01, ∗∗∗p < 0.001 (paired t test for B–D) versus Control ∗∗∗p < 0.001 (one-way ANOVA with Tukey multiple comparisons test for E and F). Red diamonds represent the averages (B–D). Scale bars, 5 μm. See also Figure S4. Image published in: Wong HH et al. (2017) © 2017 The Authors. This image is reproduced with permission of the journal and the copyright holder. This is an open-access article distributed under the terms of the Creative Commons Attribution license Larger Image Printer Friendly View

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