Supplementary Figure 9: Light controls the behavior of zebrafish embryos transiently expressing BLINK2: comparison with BLINK1 and BLINK2 Q513D.
From: A light-gated potassium channel for sustained neuronal inhibition
(a) Altered escape response in 2-d-old zebrafish embryos expressing BLINK2 (squares) or GFP (circles) (both RNAs injected at 200 pg/embryo), measured in embryos kept in the dark (black) or after 60 min of exposure to blue light (465 nm, 85 μW/mm2) (blue). The escape response was considered altered when one or two touches were not sufficient to elicit it. BLINK2 Q513D RNA was used for the experiments shown in this graph. Similar results were obtained when BLINK2 RNA was injected (data not shown); mean and s.e.m. were calculated on three (GFP) and four (BLINK2) different experiments. Total number of embryos (n) is 49 and 119, respectively. (b) BLINK120 versus BLINK2. The graph shows the number of stimulations required in order to elicit an escape response in each embryo, injected with either BLINK1 (dots) or BLINK2 (squares) RNA. BLINK2 Q513D RNA was used for the experiments shown in this graph. Measurements were performed after 30 min of exposure to blue light. The blue dashed line highlights the maximum number of tactile stimuli (8 touches) required to elicit an escape response in BLINK1 injected embryos. (c) BLINK2 versus BLINK2 Q513Q. Reversibility in dark and kinetics of the light effect on the escape response of 2-d-old embryos expressing BLINK2 (continuous line) and BLINK2 Q513D (dashed line) Blue light was turned on at time 0, after the first measurement. After 60 min of exposure, the light was turned off for 30 min and then turned on again (as indicated by blue and black bars above the graph). The response to mechanical stimulation was checked and recorded every 15 min. The total number of embryos in each group was n = 86 (BLINK2) and n = 98 (BLINK2 Q513D) from 3 independent experiments. For a, we performed two-tailed t-tests comparing GFP versus BLINK2 in the dark, GFP versus BLINK2 in blue light, GFP dark versus blue light, BLINK2 dark versus blue light. Statistically significant results are shown with asterisks (P = 0.028 and 0.0023 for BLINK2 dark/light and GFP/BLINK2 light, respectively).
