Temperature-sensitive pan mutant has defects in juvenile growth. (A) Timeline of the temperature shift experiment during zebrafish development. TS, temperate shift; wpf, weeks post-fertilization. Red bar indicates time at 34°C. (B) Whole-mount images of pan and pan/+ individuals at 8 wpf (stitched) and sde1 ts mutants, which were included as an experiment control for the effect of the restrictive temperature on regeneration mutants. Blue solid line indicates the measurement of standard length (SL). Red solid line indicates the measurement of height at anterior of anal fin (HAA). Red arrows point to severely degraded fin tissues in sde1. Scale bar: 2 mm. (C) Measurements of SL and HAA at 8 wpf (n=38 pan/+, 20 pan, 46 sde1/+, 28 sde1; mean±s.d.; two-tailed Student's t-test). (D) Timeline of a second temperature shift experiment during zebrafish development. (E) Whole-mount images of pan and pan/+ individuals at 10 wpf (stitched) and sde1 ts mutants (experiment control for the effect of the restrictive temperature on regeneration mutants). Red arrow points to degraded fin tissues in sde1. Scale bar: 2 mm. (F) Measurements of SL and HAA at 10 wpf (n=24 pan/+, 17 pan, 37 sde1/+, 18 sde1; mean±s.d.; two-tailed Student's t-test). (G) Bright-field images of pan/+ and pan at 8 wpf after a 4-week incubation at 34°C. Red arrowhead points to the growth of adult barbels. Red dashed line indicates a swim bladder. Trunk areas and tailfins in pan fish remain mostly transparent, presumably as a result of defects in the generation of adult pigmentation patterns. Scale bars: 0.5 mm (left); 1 mm (middle and right). (H) Fluorescence images of pan at 8 wpf after a 4-week incubation at 34°C. Mature calcified bone structures were detected by Alizarin Red staining. pan lacks several craniofacial bones (left, dashed lines), displays defects in scale maturation (middle, dashed lines depict scale margins) and the tailfin has no bifurcation points (right, white arrows), a landmark feature in adult tailfin bony rays. Scale bars: 0.5 mm.
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