ZFIN ID: ZDB-FIG-180621-57
González-Rosa et al., 2018 - Myocardial Polyploidization Creates a Barrier to Heart Regeneration in Zebrafish. Developmental Cell   44:433-446.e7 Full text @ Dev. Cell
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Fish:
Condition:
Anatomical Terms:
Stage: Adult
PHENOTYPE:
Fish:
Condition:
Observed In:
Stage: Adult

Fig. 1

Zebrafish Cardiomyocytes Are Mononucleated, Diploid, and Upregulate Ect2 during Heart Regeneration

(A) Schematic depicting cardiomyocyte nucleation and ploidy. See also Figure S1.

(B) Cardiomyocytes (white arrows) from dissociated Tg(cmlc2:nucGFP) hearts.

(C and D) Mononucleated and binucleated Tg(cmlc2:nucGFP) cardiomyocytes.

(E) Average percentages of mononucleated (MonoN) and binucleated (BiN) cardiomyocytes in uninjured (UI), 7-dpr, and 60-dpr ventricles (mean ± SD, n = 9,309, 9,269, and 9,578 total cells, respectively, from 4 biological replicates per group; 2 pooled ventricles per replicate; n.s., not significant by one-way ANOVA test).

(F–H) DAPI-stained mononucleated diploid, binucleated tetraploid, and mononucleated tetraploid cardiomyocytes isolated from Tg(cmlc2:nucGFP) hearts. Insets show DAPI signal.

(I) Distributions of non-cardiomyocyte (gray) and cardiomyocyte (green) DNA content in homeostatic ventricles (n = 779 and 552 cells, respectively, from 3 biological replicates; 1 ventricle per replicate). Inset is magnification to show low-frequency events (black arrowhead indicates tetraploid cardiomyocytes).

(J) Distributions of cardiomyocyte DNA content in 7-dpr and 60-dpr ventricles (blue and orange bars, respectively; overlap appears in brown). Inset is magnification to show low-frequency events (black arrowhead indicates tetraploid cardiomyocytes).

(K) Quantification of indicated cardiomyocyte populations from UI, 7-dpr, and 60-dpr ventricles (mean ± SD, n = 3,250, 2,699, and 4,306 total cells from 7, 3, and 4 biological replicates per group, respectively; 3 ventricles per replicate; ∗∗∗∗p < 0.0001; ∗∗p < 0.01 by one-way ANOVA followed by Tukey's multiple comparisons test).

(L) qPCR analysis showing relative expression of three genes involved in cytokinesis in UI, 7-dpr, and 60-dpr ventricles (mean ± SD, n = 3 technical replicates, 10 biological replicates, 1 ventricle per replicate, ∗∗∗∗p < 0.0001; ∗∗p < 0.01; p < 0.05 by one-way ANOVA followed by Tukey's multiple comparisons test). See also Figure S2.

(M and N) Single confocal sections of ect2 RNAScope (arrowheads) in UI and 7-dpr hearts. Boxed regions are shown at higher magnifications. n = 4 hearts per group with 3 sections per heart. Asterisk indicates wound edge.

CM, cardiomyocytes. Scale bars, 50 μm.

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
ect2 AB amputation: cardiac ventricle Adult regenerating tissue cardiac muscle cell RTPCR
Adult regenerating tissue cardiac muscle cell ISH
EGFP fb18Tg standard conditions Adult cardiac muscle cell IFL
kif23 AB amputation: cardiac ventricle Adult regenerating tissue cardiac muscle cell RTPCR
racgap1 AB amputation: cardiac ventricle Adult regenerating tissue cardiac muscle cell RTPCR
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
fb18Tg amputation: cardiac ventricle Adult regenerating tissue cardiac muscle cell polyploid, abnormal
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, 44, González-Rosa, J.M., Sharpe, M., Field, D., Soonpaa, M.H., Field, L.J., Burns, C.E., Burns, C.G., Myocardial Polyploidization Creates a Barrier to Heart Regeneration in Zebrafish, 433-446.e7, Copyright (2018) with permission from Elsevier. Full text @ Dev. Cell