Author response image 3. Probing the recruitment of Rohon-Beard neurons expressing GFP-Aequorin during movement.
(A, B) Lateral view showing the expression of GFP-Aequorin in 4 dpf Tg(isl2b:gal4;UAS:GFP- Aequorin-opt) larvae throughout the animal (A) and in the spinal cord (B). The arrowhead points to one Rohon-Beard cell in the dorsal spinal cord. The white dashed lines delineate the ventral and dorsal limits of the spinal cord. As previously stated for Tg(mnx1:gal4; UAS:GFP-Aequorin-opt) larvae, we verified on 5 animals that expression never targeted muscle fibers in these double transgenic animals. Ventral is bottom, rostral is left. (C) Bioluminescence signals emitted from 10 isl2b+following an acoustic stimulus during motion (blue) or after blockade of muscle contraction using pancuronium bromide in the bath (red). (D) Quantification of the amplitude of bioluminescence signals across fish in active and paralyzed larvae (4.53 +/- 0.21 versus 2.20 +/- 0.10 photons / 10 ms, n = 10 fish, n = 600 trials, p < 0.001).