a) Minimum intensity projections of a two-minute
time-lapse of fish freely swimming in a 3 cm petri dish in different
treatment conditions (n=6/7/3/3 animals in control/4-AP/TTX/4-AP+TTX, 3
independent experiments).
b) Traces of GCaMP transients Tg(elavl3:h2b-GCaMP6s)
zebrafish at 4 dpf and after overnight incubation in 0.1 mM 4-AP, and before
/ after 10 μM TTX (7 animals per conditions in 2 experiments).
c) Confocal images of Tg(mfap4:memCerulean),
Tg(olig1:nls-mApple) zebrafish at 4 dpf after treatment with 0.1 mM 4-AP,
0.5 mM 4-AP, or Danieau’s solution as control. Transmitted light
images to show spinal cord morphology and tissue integrity following drug
treatment. Scale bars: 100 μm. The graph shows that number of
macrophages which accumulate in 400 μM length of spinal cord of
Tg(mfap4:memCerulean) zebrafish after 1 day of control (2±0.25/2
cells), 0.1 mM (2±1/2 cells), and 0.5 mM (3±0.25/2 cells) 4-AP
treatment (median (25%/75% percentiles); p=0.43 (control
vs. 0.1mM 4-AP), p=0.03 (control vs. 0.5
mM 4-AP) (Kruskal-Wallis test, test statistic=3.003), n=16/19/8 animals in 3
experiments.
d) Representative images of Tg(mbp:nls-EGFP),
Tg(olig1:nls-mApple) zebrafish in control and after 2 days of 0.1 mM 4-AP
treatment (see Fig 7g for n numbers).
Scale bar: 20 μm.