ZFIN Figure: Han et al., 2019, Fig. 3

Fig. 3

Tissue-Intrinsic Effects of Vitamin D Signaling and Impact on Heart Regeneration

(A) MIP images of cyp24a1^mGFP expression in dissected hearts. Scale bar, 50 μm.

(B) MIP images of cyp24a1^mGFP expression in dissected hearts. Scale bar, 50 μm.

(C) MIP images of dissected cmlc2:FUCCI hearts in indicated transgenic backgrounds. Scale bar, 50 μm.

(D) Quantification of FUCCI⁺ CMs in (C). Mean ± SD, Mann-Whitney test and Student's t test. ^∗∗∗p 0.001; ^∗∗∗∗p < 0.0001. n = 9–10.

(E) Examples of 1 mpf cmlc2:ca-vdra and WT zebrafish. Red arrow indicates gross pericardial enlargement caused by cardiomegaly in cmlc2:ca-vdra zebrafish. Scale bar, 2 mm.

(F) Darkfield images of dissected 1 mpf hearts. a, atrium; ba, bulbus arteriosus; v, ventricle. Scale bar, 500 μm.

(G) Darkfield images of dissected livers (l) and hearts (h) from 1 mpf zebrafish. Scale bar, 500 μm.

(H) Examples of Mef2/PCNA staining of 7 dpa hearts after a single injection. Dashed lines represent amputation planes. Scale bar, 100 μm.

(I) Quantification of CM proliferation indices in (H). Mean ± SD, Mann-Whitney test. ^∗p < 0.05. n = 9.

(J) Quantified heart regeneration scores of WT or hsp70:dn-vdra(dn-vdra) zebrafish given daily heat shocks from 6 to 50 dpa. Regeneration scores (reg scores) indicate vigorous regeneration (1), partial regeneration (2), or blocked regeneration (3), as shown in Figure S3N. Two-tailed Fisher’s exact test. ^∗∗∗p < 0.001. n = 8–11.

(K) Representative images of cTnT staining at 50 dpa, after daily heat shock from 6 dpa. Scale bar, 100 μm.

(L) Representative images of AFOG stains to indicate fibrin (red) and collagen (blue) within the same hearts in (K). Arrowheadindicates scar tissue.

Expression Data

Genes:	GFP mCherry TagBFP Venus
Antibodies:	Ab1-mef2 Ab2-pcna
Fish:	WT cyp24a1^pd272Tg/+ cyp24a1^pd272Tg/+; pd301Tg/+ cyp24a1^pd272Tg/+; pd302Tg/+ pd57Tg/+; pd58Tg/+ pd301Tg/+; pd57Tg/+; pd58Tg/+ pd302Tg/+; pd57Tg/+; pd58Tg/+
Conditions:	resection: cardiac ventricle chemical treatment by injection: alfacalcidol, resection: cardiac ventricle
Anatomical Terms:	atrium cardiac muscle cell cardiac ventricle cardiac ventricle regenerating tissue heart
Stage Range:	Day 4 to Adult

Expression Detail

Antibody Labeling

Phenotype Data

Fish:	WT pd276Tg/+ pd302Tg/+ pd304Tg/+ cyp24a1^pd272Tg/+; pd301Tg/+ cyp24a1^pd272Tg/+; pd302Tg/+ pd301Tg/+; pd57Tg/+; pd58Tg/+ pd302Tg/+; pd57Tg/+; pd58Tg/+
Conditions:	heat shock, resection: cardiac ventricle chemical treatment by injection: alfacalcidol, resection: cardiac ventricle
Observed In:	atrium cardiac muscle tissue regeneration cardiac ventricle cardiac ventricle cardiac muscle cell proliferation heart liver regenerating tissue cardiac muscle cell proliferation
Stage Range:	Day 4 to Adult

Phenotype Detail

Acknowledgments

This image is the copyrighted work of the attributed author or publisher, and ZFIN has permission only to display this image to its users. Additional permissions should be obtained from the applicable author or publisher of the image.

Reprinted from Developmental Cell, 48(6), Han, Y., Chen, A., Umansky, K.B., Oonk, K.A., Choi, W.Y., Dickson, A.L., Ou, J., Cigliola, V., Yifa, O., Cao, J., Tornini, V.A., Cox, B.D., Tzahor, E., Poss, K.D., Vitamin D Stimulates Cardiomyocyte Proliferation and Controls Organ Size and Regeneration in Zebrafish, 853-863.e5, Copyright (2019) with permission from Elsevier. Full text @ Dev. Cell