Fig. 8.

Single 24 h VPA treatment prior to 78 hpf still result in developmentally delayed OTs. Short, 24 h VPA treatments were administered during three different time-windows followed by replacement of the solution with EM. Daily imaging of control (A) and 250 µM VPA-treated y304Et(cfos:Gal4); Tg(UAS:Kaede) embryos show that one 24 h VPA-treatment at 6-30 hpf (B) or 30-54 hpf (C) still leads to a smaller OT and neuropil (B. II-V, C. III-V, E,F,G) and that the development, despite the shorter treatment time, did not recuperate. (D) When the VPA-treatment was applied between 54-78 hpf, OT development was affected less than in previous treatments (B,C). (E, F,G) Three embryos per experimental condition were imaged and quantified (except for the 6-30 hpf group, where the image from only one embryo prevented analysis at 96 hpf and 120 hpf). At 48 hpf, length, width, and depth are difficult to measure since during this time neuroepithelium is transitioning into neuroblasts. A two-way ANOVA with Šidák's multiple comparisons test was performed in E-G *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001. Data are shown as mean±s.e.m.