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. Author manuscript; available in PMC: 2022 Sep 1.
Published in final edited form as: Dev Dyn. 2020 Oct 26;250(9):1330–1339. doi: 10.1002/dvdy.261

Figure 2. Inhibition of vitamin D receptor signaling impairs fin regeneration.

Figure 2.

(A) In situ hybridization experiments for expression of cyp24a1, a VDR target gene, or cyp26a1, an RAR target gene. Embryos microinjected with 25 ng/μL dn-vdra mRNA at the single-cell stage displayed visibly reduced cyp24a1, but not cyp26a1, expression level at 24 hpf. n = 20 for each group. (B, C) Whole-animal induction of dominant-negative VDR in transgenic fish significantly delayed fin regeneration at 4 and 7 dpa. n = 8 for each group. (D-G) Transient, local inhibition of vitamin D signaling using regeneration-responsive lepb regulatory elements to induce cyp24a1 (D, F) or dn-vdra (E, G) impairs fin regeneration. In cyp24a1 overexpression studies, n = 13 for wild-type fish at 4 dpa; n = 12 for wild-types at 7 dpa; n = 11 for transgenic fish at both time points. In dn-vdra overexpression groups, n = 11 for wild-types; n = 13 for transgenic fish. (H) Fin regeneration was modestly delayed by 7 dpa in vdra/vdrb double knockout zebrafish. Values on right incorporate corrections for the tail fin sizes of heterozygous and (the smaller) mutant animals, as explained in the main text, which is why relative lengths are plotted. Values in (H) were normalized to the heterozygous 7 dpa average lengths (set at 1.0). n = 14 for each group. Dashed lines indicate approximate amputation planes.