ZFIN ID: ZDB-FIG-190716-2
Han et al., 2019 - Vitamin D Stimulates Cardiomyocyte Proliferation and Controls Organ Size and Regeneration in Zebrafish. Developmental Cell   48(6):853-863.e5 Full text @ Dev. Cell
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Gene:
Antibody:
Fish:
Conditions:
Anatomical Terms:
Stage Range: Protruding-mouth to Day 4
PHENOTYPE:
Fish:
Conditions:
Observed In:
Stage Range: Protruding-mouth to Adult

Fig. 2

Vitamin D Signaling Enhances Organismal Size

(A) MIP images and quantification of H3P staining of 3 dpf embryos. hpt, hours post-treatment; H3P, phospho-Histone H3. Mean ± SD, Mann-Whitney test and Student's t test. ∗∗∗∗p < 0.0001. n = 11–13. Scale bar, 500 μm.

(B) Example and quantification of 4 dpf zebrafish growth. Mean ± SD, Mann-Whitney test and Student’s t test. ∗∗p < 0.01; ∗∗∗p < 0.001. n = 12-14. Scale bar, 500 μm.

(C) Growth and rescue of 2 mpf juvenile wild-type (WT) and ubb:cyp24a1 zebrafish. Scale bar, 4 mm.

(D) Quantification of zebrafish length before and after treatment. Mean ± SD. n = 14–15.

(E) mGFP and TagBFP expression in 4 dpf embryos treated with vehicle or 10 nM Alfa. Scale bar, 500 μm.

(F) Quantified body length of two independent ubb:cyp24a1 lines and WT siblings at 4 dpf. Mean ± SD, Mann-Whitney test. p < 0.05. n = 14–23.

(G) cyp24a1mGFP expression in 1 μM Alfa-treated embryos.

(H) Quantification of relative growth of 4 dpf embryos treated with vehicle or Alfa for 24 h. Mean ± SD, Student’s t test. n = 13.

(I) Representative images and body length quantification of 6 mpf zebrafish. mpf, months post-fertilization. Mean ± SD, Student's t test. n = 7. Scale bar, 5 mm.

(J) mGFP and TagBFP expression of 3 dpf embryos after a single heat-shock and 24-h drug treatment. Scale bar, 500 μm.

(K) Quantified relative growth of 4 dpf embryos after a heat shock and 24-h drug treatment. Mean ± SD, Student’s t test. n = 10.

Gene Expression Details
Gene Antibody Fish Conditions Stage Qualifier Anatomy Assay
GFP cyp24a1pd272Tg/+ control Protruding-mouth heart IFL
Day 4 heart IFL
cyp24a1pd272Tg/+ chemical treatment by environment: alfacalcidol Protruding-mouth heart IFL
Protruding-mouth liver IFL
Protruding-mouth whole organism IFL
Day 4 heart IFL
Day 4 liver IFL
cyp24a1pd272Tg/+; pd273Tg/+ control Day 4 Not Detected heart IFL
cyp24a1pd272Tg/+; pd273Tg/+ chemical treatment by environment: alfacalcidol Day 4 heart IFL
Day 4 liver IFL
cyp24a1pd272Tg/+; pd276Tg/+ heat shock Protruding-mouth heart IFL
cyp24a1pd272Tg/+; pd276Tg/+ heat shock, chemical treatment by environment: alfacalcidol Protruding-mouth heart IFL
Protruding-mouth liver IFL
vdrapd307/pd307; vdrbpd308/pd308; cyp24a1pd272Tg/+ chemical treatment by environment: alfacalcidol Day 4 Not Detected heart IFL
Day 4 Not Detected whole organism IFL
Antibody Labeling Details
Antibody Assay Fish Conditions Stage Anatomy
Ab19-h3 IHC WT control Protruding-mouth trunk
IHC WT chemical treatment by environment: alfacalcidol Protruding-mouth trunk
Phenotype Details
Fish Conditions Stage Phenotype
WT chemical treatment by environment: alfacalcidol Protruding-mouth trunk ab19-h3 labeling increased amount, abnormal
Protruding-mouth trunk cell population proliferation increased occurrence, abnormal
Protruding-mouth - Day 4 growth increased rate, abnormal
Day 4 growth increased rate, abnormal
Day 4 whole organism increased length, abnormal
Adult growth increased rate, abnormal
Adult whole organism increased length, abnormal
WT chemical treatment by environment: calcipotriol Protruding-mouth - Day 4 growth increased rate, abnormal
Day 4 whole organism increased length, abnormal
cyp24a1pd272Tg/+ chemical treatment by environment: alfacalcidol Protruding-mouth heart GFP expression increased amount, abnormal
Protruding-mouth liver GFP expression increased amount, abnormal
Protruding-mouth whole organism GFP expression increased amount, abnormal
Day 4 heart GFP expression increased amount, abnormal
Day 4 liver GFP expression increased amount, abnormal
cyp24a1pd272Tg/+; pd273Tg/+ control Day 4 heart GFP expression absent, abnormal
cyp24a1pd272Tg/+; pd273Tg/+ chemical treatment by environment: alfacalcidol Day 4 heart GFP expression amount, ameliorated
Day 4 liver GFP expression amount, ameliorated
cyp24a1pd272Tg/+; pd276Tg/+ heat shock Protruding-mouth heart GFP expression decreased amount, abnormal
cyp24a1pd272Tg/+; pd276Tg/+ heat shock, chemical treatment by environment: alfacalcidol Protruding-mouth heart GFP expression amount, ameliorated
Protruding-mouth liver GFP expression amount, ameliorated
pd273Tg/+ control Adult growth decreased rate, abnormal
Adult whole organism decreased length, abnormal
pd273Tg/+ chemical treatment by environment: alfacalcidol Adult growth rate, ameliorated
Adult whole organism length, ameliorated
pd274Tg/+ control Adult growth decreased rate, abnormal
Adult whole organism decreased length, abnormal
pd274Tg/+ chemical treatment by environment: alfacalcidol Adult growth rate, ameliorated
Adult whole organism length, ameliorated
pd276Tg/+ heat shock Protruding-mouth growth decreased rate, abnormal
pd276Tg/+ heat shock, chemical treatment by environment: alfacalcidol Protruding-mouth growth decreased rate, abnormal
vdrapd307/pd307 ; vdrbpd308/pd308 control Adult whole organism decreased length, abnormal
vdrapd307/pd307 ; vdrbpd308/pd308 chemical treatment by environment: alfacalcidol Day 4 growth rate, ameliorated
Day 4 whole organism length, ameliorated
vdrapd307/pd307; vdrbpd308/pd308; cyp24a1pd272Tg/+ chemical treatment by environment: alfacalcidol Day 4 heart GFP expression absent, abnormal
Day 4 whole organism GFP expression absent, abnormal
Acknowledgments:
ZFIN wishes to thank the journal Developmental Cell for permission to reproduce figures from this article. Please note that this material may be protected by copyright.

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