Figure 1.

Chemical screening protocol for modulators of hair cell regeneration. A, Schematic diagramming the regeneration screen. Five to 6 dpf Tg(pou4f3:gap43-GFP) zebrafish expressing membrane-bound GFP under the pou4f3 promoter were treated with 400 μm neomycin for 1 h, rinsed and transferred into 48-well plates. Fish were then either treated for 48 h with a test drug, DMSO vehicle, or DAPT (a positive control for enhanced hair cell regeneration) and subsequently scored for GFP+ hair cells on a 1–5 scale, with a score of 3 representing normal regeneration. B, Fluorescent images of the MI₁ neuromasts illustrate the scoring scale used in the screen for regeneration modulators. Examples from Tg(pou4f3:gap43-GFP) larval zebrafish that would have received a score of 1, 3, and 5 are shown. Scale bar, 10 μm.