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Figure 5. Local Synthesis of β-actin Is Required for Axon Branching In Vivo(A and B) Axon branching in the tectum after local delivery of MO at stages 41–43. More protrusions were added than removed in Control MO (Con MO) condition (filopodia: t17 = 3.1, p = 0.007; branches: t17 = 2.4, p = 0.03) (A′ and A″). No significant differences were observed in the number of protrusions that were added and removed in β-actin MO (β-aMO) condition (addition: t25 = 1.4, p = 0.16; removal: t25 = 1.9, p = 0.07) (B′ and B″).(C) Filopodia dynamics were inhibited in β-aMO condition (addition: t42 = 3.9, p = 0.0004; removal: t42 = 2.7, p = 0.01).(D) Branch dynamics were inhibited in β-aMO condition (addition: t42 = 3.1, p = 0.004; removal: t42 = 3.0, p = 0.005).(E) Local delivery of MO into RGC axons by tectum electroporation and imaged immediately thereafter. Electroporated eye was removed to eliminate somatic contribution.(F and G) Time-lapse images of axonal branching in the tectum at stages 41–43 after local delivery of MO at stages 35/36–37/38. More protrusions were added than removed in Con MO condition (filopodia: t11 = 3.4, p = 0.006; branches: t11 = 4.9, p = 0.0005) (F′ and F″). More protrusions were added than removed in β-aMO condition (addition: t17 = 4.2, p = 0.0006; removal: t17 = 4.0, p = 0.0008) (G′ and G″).(H and I) The dynamics of filopodia (H; addition: t28 = 0.6, p = 0.58; removal: t28 = 0.6, p = 0.55) and branches (I; addition: t28 = 0.3, p = 0.78; removal: t28 = 0.07, p = 0.95) were unaffected in β-aMO condition.(J) Local delivery of MO into the tectum before tectal entry of RGC axons (stages 35/36–37/38) and live imaging of axonal branching after tectal entry of RGC axons (stages 41–43). Scissors and dashed line denote that only the skin overlying the tectal area was removed to minimize damage to the brain.Error bars represent SEM. ∗p < 0.01, ∗∗p < 0.01, ∗∗∗p < 0.001 (paired t test for A, B, F, and G and unpaired t test for C, D, H, and I). Scale bars, 5 μm. Red diamonds represent the averages (A, B, F, and G). See also Figures S4 and S6. 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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