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ZFIN ID: ZDB-FIG-170201-25
Rydeen et al., 2016 - Cyp26 Enzymes Facilitate Second Heart Field Progenitor Addition and Maintenance of Ventricular Integrity. PLoS Biology   14:e2000504 Full text @ PLoS Biol.
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Antibodies:
Fish:
Knockdown Reagents:
Anatomical Terms:
Stage: Long-pec
PHENOTYPE:
Fish:
Knockdown Reagents:
Observed In:
Stage: Long-pec

Fig. 3

Ventricular cardiomyocytes can exit the heart tube in Cyp26-deficient embryos.

(A,B) 30-min interval frames from confocal time-lapse movies of control and Cyp26-deficient Tg(myl7:EGFP) hearts. Images of hearts were depth-coded with the spectrum ranging from pink at 120 μm to blue at 0 μm. (C,D) Hematoxylin–eosin (HE) stained frontal sections of the hearts from control and Cyp26-deficient embryos. Endocardium (white arrowheads) and myocardium (black arrowheads). Control hearts n = 4. Cyp26-deficient hearts n = 5. (E,F) Control and Cyp26-deficient Tg(myl7:EGFP);Tg(kdrl:mCherry) embryos at 48 hpf. Endocardium (red) and myocardium (green). Arrowheads indicate the inner border of endocardial and outer border of myocardial cells. (G) Graph showing the distance between the endocardial and myocardial layers (control n = 5, Cyp26 deficient n = 7). (H–I”‘) Confocal images of control and Cyp26-deficient Tg(myl7:EGFP) hearts stained for zonula occludens 1 (ZO1) (red) and green fluorescent protein (GFP) (green), with schematized outlines of cell boundaries and ZO1 staining. Arrow denotes cardiomyocyte protruding into the pericardial space. (J) Graph depicting the percentage of ZO1 expression along the height of cardiomyocytes (control n = 15, Cyp26 deficient n = 15). (K) Graph depicting circularity measurement of ventricular cells (control n = 15, Cyp26 deficient n = 30). (L–M''') Confocal images of control and Cyp26-deficient Tg(myl7:EGFP) hearts stained for β-catenin (red) and GFP (green), with schematized outlines of cell boundaries and β-catenin staining. Error bars are SEM, asterisk denotes p < 0.05 by Student’s t test. Frontal views, anterior up (A–F,H,I,L,M); n > 20 embryos per group (E,F,H,I,L,M). A, apical; B, basal. Scale bar for C–M’: 50 μm. Scale bar for H''–M''': 25 μm.

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
EGFP twu34Tg control Long-pec ventricular myocardium IHC
twu34Tg + MO1-cyp26c1 + MO2-cyp26c1 + MO4-cyp26a1 + MO5-cyp26a1 control Long-pec ventricular myocardium IHC
twu34Tg ; ci5Tg control Long-pec ventricular myocardium IHC
twu34Tg ; ci5Tg + MO1-cyp26c1 + MO2-cyp26c1 + MO4-cyp26a1 + MO5-cyp26a1 control Long-pec ventricular myocardium IHC
mCherry twu34Tg ; ci5Tg control Long-pec ventricular endocardium IHC
twu34Tg ; ci5Tg + MO1-cyp26c1 + MO2-cyp26c1 + MO4-cyp26a1 + MO5-cyp26a1 control Long-pec ventricular endocardium IHC
Antibody Labeling Details
Antibody Assay Fish Conditions Stage Anatomy
Ab1-ctnnb IHC twu34Tg control Long-pec ventricular myocardium cell-cell junction
IHC twu34Tg + MO1-cyp26c1 + MO2-cyp26c1 + MO4-cyp26a1 + MO5-cyp26a1 control Long-pec ventricular myocardium cell-cell junction
Ab1-tjp1 IHC twu34Tg control Long-pec ventricular myocardium bicellular tight junction
IHC twu34Tg + MO1-cyp26c1 + MO2-cyp26c1 + MO4-cyp26a1 + MO5-cyp26a1 control Long-pec ventricular myocardium bicellular tight junction
Phenotype Details
Fish Conditions Stage Phenotype
twu34Tg + MO1-cyp26c1 + MO2-cyp26c1 + MO4-cyp26a1 + MO5-cyp26a1 control Long-pec cardiac ventricle cardiac muscle cell circular, abnormal
Long-pec cardiac ventricle cardiac ventricle morphogenesis decreased process quality, abnormal
Long-pec cardiac ventricle establishment of cell polarity decreased process quality, abnormal
Long-pec ventricular myocardium bicellular tight junction ab1-tjp1 labeling spatial pattern, abnormal
Long-pec ventricular myocardium cardiac muscle cell mislocalised, abnormal
Long-pec ventricular myocardium cell-cell junction ab1-ctnnb labeling spatial pattern, abnormal
twu34Tg ; ci5Tg + MO1-cyp26c1 + MO2-cyp26c1 + MO4-cyp26a1 + MO5-cyp26a1 control Long-pec cardiac ventricle cardiac ventricle morphogenesis decreased process quality, abnormal
Long-pec ventricular myocardium increased distance ventricular endocardium, abnormal
Acknowledgments:
ZFIN wishes to thank the journal PLoS Biology for permission to reproduce figures from this article. Please note that this material may be protected by copyright. Full text @ PLoS Biol.