Fig. 7.
In vivo inhibition of coiling activity of zebrafish using fused-bPAC-K. a Schematic representation of the construct injected to generate the UAS:bPAC-SthK-tRPFtmpn157 transgenic line. The transgenic zebrafish expresses a fusion protein of bPAC(wt), SthK, and tagRFPt under the control of UAS. b Expression of bPAC-SthK-tRFPt in the spinal cord of a 24 h post-fertilization Gal4s1020t, UAS:bPAC-Sthk-tRFPtmpn157 embryo. Scale bar: 50 µm. c High magnification of the spinal cord in fused-bPAC-K transgenic zebrafish. Scale bar: 10 µm. d–f Raster plots of coiling events in 25 fused-bPAC-K transgenic embryos (black) and 10 non-expressing embryos from the same clutch (gray) before, during, and after illumination with blue or red light. g–i Average number of coils per 5 s over three replicates in fused-bPAC-SthK-positive (black) and non-expressing (gray) embryos. All zebrafish embryos show low-level photoinhibition of motor responses due to endogenous light sensitivity. Error bars represent SEM