ZFIN ID: ZDB-FIG-180208-10
Donat et al., 2018 - Heg1 and Ccm1/2 proteins control endocardial mechanosensitivity during zebrafish valvulogenesis. eLIFE   7:e28939 Full text @ Elife
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Antibody:
Fish:
Anatomical Terms:
Stage Range: Long-pec to Day 4
PHENOTYPE:
Fish:
Observed In:
Stage Range: Long-pec to Day 4

Fig. 3

Krit1 is required for abluminal cell fates during valvulogenesis.

(A–F) Single confocal z-section images of endocardial cells within the atrioventricular canal (AVC) region marked by Tg(kdrl:EGFP)s843 expression (cyan) and Alcam staining (magenta). (A) At 48 hpf, wild-type (WT) endocardial cushion cells express Alcam (asterisks). (B) In krit1ty219c mutants, endocardial cells of the AVC region do not express Alcam and cushions do not form (arrow). (C) Injection of egfp-krit1 mRNA rescues cardiac cushion formation in krit1ty219c mutants (n = 15/15). krit1ty219c mutant hearts form cardiac cushions and endocardial cushion cells express Alcam. (D) By 96 hpf, Alcam expression is mainly restricted to luminal endocardial cells of the developing WT cardiac valve leaflet (asterisks). (E) krit1ty219c mutants do not form valve leaflets but AVC endocardial cells express Alcam (asterisks). (F) krit1ty219c mutant that was initially rescued by egfp-krit1 mRNA injection has a dysmorphic cardiac valve leaflet at 96 hpf with an agglomeration of luminal Alcam-positive cells (asterisks). (G–L) Whole-mount in situ hybridization of klf2a cardiac expression. (G) At 54 hpf, klf2a expression is restricted to endocardial cells of the AVC in WT while (H) klf2a is strongly expressed throughout the entire endocardium in krit1ty219c mutant. (I) krit1ty219c mutant rescued by injection of egfp-krit1 mRNA has a restricted and strong klf2a expression at the AVC (n = 6/6). (J) By 96 hpf, klf2a expression is restricted to the AVC in WT while (K) klf2a is strongly expressed throughout the entire endocardium in krit1ty219c mutants. (L) krit1ty219c mutant embryo injected with egfp-krit1 mRNA has high klf2a levels throughout the entire endocardium (n = 8/8). A: atrium, V: ventricle, L: luminal, AL: abluminal. Scale bars are 10 μm.

Gene Expression Details
Gene Antibody Fish Conditions Stage Qualifier Anatomy Assay
alcama zn-8 krit1ty219c/ty219c ; s843Tg standard conditions Long-pec Not Detected atrioventricular canal endocardium IHC
zn-8 Day 4 atrioventricular canal endocardium IHC
zn-8 s843Tg standard conditions Long-pec atrioventricular canal endocardial cushion IHC
zn-8 Day 4 atrioventricular valve endocardium IHC
EGFP krit1ty219c/ty219c ; s843Tg standard conditions Long-pec atrioventricular canal endocardium IHC
Long-pec endocardium IHC
Day 4 endocardium IHC
s843Tg standard conditions Long-pec atrioventricular canal endocardial cushion IHC
Long-pec endocardium IHC
Day 4 endocardium IHC
klf2a WT standard conditions Long-pec Not Detected atrial endocardium ISH
Long-pec atrioventricular canal endocardium ISH
Long-pec Not Detected ventricular endocardium ISH
Day 4 Not Detected atrial endocardium ISH
Day 4 atrioventricular canal endocardium ISH
Day 4 Not Detected ventricular endocardium ISH
krit1ty219c/ty219c standard conditions Long-pec atrial endocardium ISH
Long-pec endocardium ISH
Long-pec ventricular endocardium ISH
Day 4 atrial endocardium ISH
Day 4 endocardium ISH
Day 4 ventricular endocardium ISH
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
krit1ty219c/ty219c standard conditions Long-pec atrial endocardium klf2a expression mislocalised, abnormal
Long-pec endocardium klf2a expression increased distribution, abnormal
Long-pec ventricular endocardium klf2a expression mislocalised, abnormal
Day 4 atrial endocardium klf2a expression mislocalised, abnormal
Day 4 endocardium klf2a expression increased amount, abnormal
Day 4 endocardium klf2a expression increased distribution, abnormal
Day 4 ventricular endocardium klf2a expression mislocalised, abnormal
krit1ty219c/ty219c ; s843Tg standard conditions Long-pec atrioventricular canal endocardial cushion formation arrested, abnormal
Long-pec atrioventricular canal endocardium alcama expression absent, abnormal
Long-pec atrioventricular canal lacks all parts of type atrioventricular canal endocardial cushion, abnormal
Day 4 atrioventricular canal endocardial cushion formation arrested, abnormal
Day 4 atrioventricular canal lacks all parts of type atrioventricular canal endocardial cushion, abnormal
Acknowledgments:
ZFIN wishes to thank the journal eLIFE for permission to reproduce figures from this article. Please note that this material may be protected by copyright. Full text @ Elife