header logo image header logo text
Downloads Login
Research
General Information
ZIRC
ZFIN ID: ZDB-FIG-180417-29
Rogers et al., 2017 - Nodal patterning without Lefty inhibitory feedback is functional but fragile. eLIFE   6 Full text @ Elife
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Fish:
Condition:
Anatomical Terms:
Stage: 75%-epiboly
PHENOTYPE:
Fish:
Condition:
Observed In:
Stage Range: 75%-epiboly to Prim-5

Fig. 6

Artificial, uniform Nodal inhibition rescues Lefty loss.

Wild type and lft1-/-;lft2-/- mutant embryos were exposed to 0, 1.25, 2.5, 3.75, 5, or 6.25 μM SB-505124 (Nodal inhibitor drug) starting at the 8 cell stage, sphere stage, or shield stage. (Exposure to 30–50 μM fully inhibits Nodal signaling (Fan et al., 2007; Hagos et al., 2007; Hagos and Dougan, 2007).) Drug was removed at 24 hr post-fertilization (hpf). (A) Inhibition is decoupled from signaling in drug-treated lft1-/-;lft2-/- mutants. (B–F) Exposure to 2.5 μM SB-505124 at the 8 cell (D) or sphere (E) stage or 5 μM at shield stage (F) rescues gross morphological defects in lft1-/-;lft2-/- mutants (C); compare to wild type embryo (B). Embryos were imaged at 1 day post-fertilization (dpf). (G,H) Drug exposure rescues lft1-/-;lft2-/- mutants to adulthood. (G) 10 month old wild type TE adult male. (H) 5 month old lft1-/-;lft2-/- mutant male treated with 3.5 μM SB-505124 from sphere stage to 24 hpf. (I) Percent of wild type (black bars) or double mutant (gray bars) embryos with normal gross morphology at 1 dpf after drug exposure is plotted. See Figure 6—figure supplements 1 and 2 for further details. (J–U’’) Endodermal (sox32/casanova) and mesodermal (noto/floating head) gene expression in embryos treated as indicated was assessed using in situ hybridization. lft1-/-;lft2-/- mutants treated starting at the 8 cell stage (2.5 μM) exhibit decreases in endoderm compared to wild type embryos (J–L’’), but have mesodermal gene expression similar to wild type embryos (M–O’’). lft1-/-;lft2-/- mutants treated at shield stage (5 μM) exhibit upregulation of mesendoderm until ~5 hr post-exposure (P–U’’). See Figure 6—figure supplement 3 for in situ hybridization of embryos treated with drug at sphere and shield stage, as well as additional time points and treatments.

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
noto WT standard conditions 75%-epiboly axis ISH
lft1a145/a145 ; lft2a146/a146 standard conditions 75%-epiboly axis ISH
lft1a145/a145 ; lft2a146/a146 chemical treatment by environment: SB 505124 75%-epiboly axis ISH
sox32 WT standard conditions 75%-epiboly endoderm ISH
lft1a145/a145 ; lft2a146/a146 standard conditions 75%-epiboly endoderm ISH
lft1a145/a145 ; lft2a146/a146 chemical treatment by environment: SB 505124 75%-epiboly endoderm ISH
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
lft1a145/a145 ; lft2a146/a146 standard conditions 75%-epiboly axis noto expression increased amount, abnormal
75%-epiboly axis noto expression increased distribution, abnormal
75%-epiboly axis noto expression spatial pattern, abnormal
75%-epiboly endoderm sox32 expression increased amount, abnormal
75%-epiboly endoderm sox32 expression increased distribution, abnormal
75%-epiboly endoderm sox32 expression spatial pattern, abnormal
Prim-5 caudal fin decreased length, abnormal
Prim-5 caudal fin morphology, abnormal
Prim-5 eye aplastic, abnormal
Prim-5 eye morphology, abnormal
Prim-5 whole organism decreased length, abnormal
Prim-5 whole organism morphology, abnormal
Prim-5 whole organism necrotic, abnormal
lft1a145/a145 ; lft2a146/a146 chemical treatment by environment: SB 505124 75%-epiboly axis noto expression amount, ameliorated
75%-epiboly axis noto expression spatial pattern, ameliorated
75%-epiboly endoderm sox32 expression amount, ameliorated
75%-epiboly endoderm sox32 expression spatial pattern, ameliorated
Prim-5 caudal fin length, ameliorated
Prim-5 caudal fin morphology, ameliorated
Prim-5 eye morphology, ameliorated
Prim-5 whole organism morphology, ameliorated
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