ZFIN ID: ZDB-FIG-200319-5
Marín-Juez et al., 2019 - Coronary Revascularization During Heart Regeneration Is Regulated by Epicardial and Endocardial Cues and Forms a Scaffold for Cardiomyocyte Repopulation. Developmental Cell   51:503-515.e4 Full text @ Dev. Cell
ADDITIONAL FIGURES
EXPRESSION / LABELING:
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Conditions:
Anatomical Terms:
Stage: Adult
PHENOTYPE:
Fish:
Condition:
Observed In:
Stage: Adult

Fig. 1 Regenerating Coronaries Sprout Superficially and toward the Cardiac Lumen (A) cEC proliferation at different times after injury (n = 4 ventricles per time point). (B) Ventricle at 7 dpci stained for coronaries (green), CMs (red), and DNA (blue); (Bi, Bii) high-magnification images of coronary vessels sprouting into the ventricle (white arrowheads) and superficially (magenta arrowheads). (C and D) 3D image reconstructions of the injured area after tissue clearing and staining for coronaries (red) and CMs (green). (C) Luminal view of intra-ventricular coronary sprouting (arrowheads) in the border zone (white arrows). Sprouts (yellow arrowheads) shown in high magnification (n = 3). (D) Intra-ventricular sprouting vessels in the apex (arrowheads). High-magnification images of sprouts (yellow arrowheads) shown in i and ii (n = 3). Asterisk marks the border zone. (E) Tg(-0.8flt1:RFP); TgBAC(apln:EGFP) ventricle at 96 hpci. Yellow dotted line delineates the area with apln:EGFP+ coronaries (n = 6). (F) Ventricular section at 96 hpci stained for coronaries (red), apln:EGFP expression (green), PCNA (white), and DNA (blue). High-magnification images of coronary vessels sprouting intra-ventricularly (Fi, white arrowheads) and superficially (Fii, white arrowheads) (n = 6). (G) qPCR analysis of ppargc1a, apln, ndufb5, and atp5j mRNA levels in sorted -0.8flt1:RFP+ cECs at 96 h post-sham (hps) (n = 4) and 96 hpci (n = 4). (H) qPCR analysis of ppargc1a, apln, ndufb5, and atp5j mRNA levels in sorted -0.8flt1:RFP+ cECs from 8 months post-fertilization (mpf) untouched WT (n = 4) and ppargc1a−/− (n = 4) ventricles. (I) qPCR analysis of ppargc1a, apln, ndufb5, and atp5j mRNA levels in sorted -0.8flt1:RFP+ cECs from 8 months post-fertilization (mpf) WT (n = 4) and ppargc1a−/− (n = 4) ventricles at 96 hpci. (J) cEC proliferation in 8 mpf WT (n = 5), ppargc1a−/− (n = 5), and apln−/− (n = 5) ventricles. (K) Section of a TgBAC(apln:CreERT2); Tg(ubb:GSR) ventricle at 7 dpci stained for GFP (white), mCherry (green), CMs (red), and DNA (blue); right panel shows high-magnification image of recombined coronaries (white arrows) (n = 6). White dotted lines delineate injured area. Data in graphs expressed as mean ± SEM. ns, no significant difference, ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001. Scale bars: 50 μm. See also Figures S1–S3; Videos S1 and S2.

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
apln hu5333Tg cryoablation: heart Adult coronary artery endothelial cell RTPCR
ppargc1abns176/bns176; hu5333Tg control Adult coronary artery endothelial cell RTPCR
ppargc1abns176/bns176; hu5333Tg cryoablation: heart Adult coronary artery endothelial cell RTPCR
atp5pf hu5333Tg cryoablation: heart Adult coronary artery endothelial cell RTPCR
ppargc1abns176/bns176; hu5333Tg control Adult coronary artery endothelial cell RTPCR
ppargc1abns176/bns176; hu5333Tg cryoablation: heart Adult coronary artery endothelial cell RTPCR
EGFP bns157Tg; hu5333Tg cryoablation: heart Adult regenerating tissue coronary artery IHC
mCherry bns310Tg; cz1701Tg cryoablation: heart, chemical treatment by injection: afimoxifene Adult coronary artery endothelial cell IHC
Adult regenerating tissue coronary artery IHC
ndufb5 ppargc1abns176/bns176; hu5333Tg control Adult coronary artery endothelial cell RTPCR
ppargc1abns176/bns176; hu5333Tg cryoablation: heart Adult coronary artery endothelial cell RTPCR
ppargc1a hu5333Tg cryoablation: heart Adult coronary artery endothelial cell RTPCR
ppargc1abns176/bns176; hu5333Tg control Adult coronary artery endothelial cell RTPCR
ppargc1abns176/bns176; hu5333Tg cryoablation: heart Adult coronary artery endothelial cell RTPCR
RFP bns157Tg; hu5333Tg cryoablation: heart Adult regenerating tissue coronary artery IHC
hu5333Tg cryoablation: heart Adult coronary artery IHC
Adult regenerating tissue coronary artery IHC
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
aplnmu267/mu267; hu5333Tg cryoablation: heart Adult blood vessel endothelial cell cell population proliferation decreased occurrence, abnormal
Adult regenerating tissue cell population proliferation decreased occurrence, abnormal
hu5333Tg cryoablation: heart Adult blood vessel endothelial cell cell population proliferation increased occurrence, abnormal
Adult coronary artery endothelial cell apln expression increased amount, abnormal
Adult coronary artery endothelial cell atp5pf expression decreased amount, abnormal
Adult coronary artery endothelial cell ppargc1a expression decreased amount, abnormal
ppargc1abns176/bns176; hu5333Tg control Adult coronary artery endothelial cell apln expression increased amount, abnormal
Adult coronary artery endothelial cell atp5pf expression decreased amount, abnormal
Adult coronary artery endothelial cell ndufb5 expression decreased amount, abnormal
Adult coronary artery endothelial cell ppargc1a expression decreased amount, abnormal
ppargc1abns176/bns176; hu5333Tg cryoablation: heart Adult blood vessel endothelial cell cell population proliferation increased occurrence, abnormal
Adult coronary artery endothelial cell apln expression increased amount, abnormal
Adult coronary artery endothelial cell atp5pf expression decreased amount, abnormal
Adult coronary artery endothelial cell ndufb5 expression decreased amount, abnormal
Adult coronary artery endothelial cell ppargc1a expression decreased amount, abnormal
Adult regenerating tissue cell population proliferation increased occurrence, 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, 51, Marín-Juez, R., El-Sammak, H., Helker, C.S.M., Kamezaki, A., Mullapuli, S.T., Bibli, S.I., Foglia, M.J., Fleming, I., Poss, K.D., Stainier, D.Y.R., Coronary Revascularization During Heart Regeneration Is Regulated by Epicardial and Endocardial Cues and Forms a Scaffold for Cardiomyocyte Repopulation, 503-515.e4, Copyright (2019) with permission from Elsevier. Full text @ Dev. Cell