ZFIN Image: Halabi et al., 2022, Fig. 4

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Figure Caption

Fig. 4

Ventricle and atrium are smaller in sema3fb mutant embryos. Whole mount ISH of 48 hpf hearts. Dotted yellow lines mark the presumptive border between the atria and ventricle placed at the morphological atrioventricular constriction. A-C The cardiomyocyte marker tnnt2a reveals large chambers in the WT heart (N = 2, n = 16/18) (A), while both sema3fb mutant alleles present with small hearts (B, N = 2, 14/16; C, N = 2, 15/16). D Antero-posterior heart length (purple line, schematic) reveals smaller sema3fb^ca305 (p < 0.0001) and sema3fb^ca306 (p = 0.0016) hearts as compared to WT. E–G Mutant alleles (F,G) present with smaller myh7 + ventricles (N = 2, 17/20 sema3fb^ca305; N = 2, 15/18 sema3fb^ca306) than WT (N = 2, n = 19/20) (E), with loss of myh7 label at the AVC (yellow arrowheads). H Shorter ventricles (purple line, schematic) in sema3fb^ca305 (p = 0.009) and sema3fb^ca306 (p = 0.013) as compared to WT embryos. I-K Mutant alleles present with smaller myh6 + atria (N = 2, 18/21 sema3fb^ca305; N = 2, 15/17 sema3fb^ca306) (J,K) than in WT (N = 2, n = 19/19) (I), and ectopic myh6 label in the ventricles (yellow arrowheads). L Quantitation of area (purple fill) reveals smaller atria in sema3fb^ca305 (p = 0.0001) and sema3fb^ca306 (p < 0.0001) hearts as compared to WT. M, N myh6 expression at 72 hpf reveals a smaller sema3fb^ca305 (N = 2, n = 27) atrium as compared to WT (n = 18). O-R Expression of irx1a (O-P; N = 2; WT n = 15/15; sema3fb^ca305 n = 22/22) and tbx5a (Q-R; N = 2; WT n = 15/15; sema3fb^ca305 n = 15/15) in the ventricle, with arrows marking the morphological atrioventricular constriction. Insets (O,P) reveal the irx1a border at the AVC, and the dotted lines the length of the irx1a domain border. S Mean area of myh6 ISH label at 72 hpf (p = 0.0021). T Mean myh7(-) area measured between the morphological AVC and the posterior border of high myh7 expression (p < 0.0001). U, V myh7 ISH to mark ventricular cardiomyocytes and S46 antibody to label atrial cardiomyocytes. In the 48 hpf sema3fb^ca305 heart there is a gap between the two labels (bar) into which S46 immunolabel extends (arrows). A: anterior; a, atrium; P: posterior; v, ventricle. Scale bar 50 µm

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Acknowledgments

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