ZFIN ID: ZDB-FIG-180426-14
Bisgrove et al., 2017 - Maternal Gdf3 is an obligatory cofactor in nodal signaling for embryonic axis formation in zebrafish. eLIFE   6 Full text @ Elife
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Fish:
Anatomical Terms:
Stage: Prim-5
PHENOTYPE:
Fish:
Observed In:
Stage: Prim-5

Fig. 1 Zygotic gdf3 mutants are viable and maternal zygotic mutants have phenotypes indicative of loss of Nodal signaling.

(A–C) gdf3 mutants were generated using TALEN-mediated mutagenesis. (A) Gdf3, a TGFβ family member, comprises a signal peptide, pro-domain and mature TGFβ domain. TALENs were designed to target genomic sequences located near the amino end of the pro-domain. (B) Mutants were identified by high-resolution melt analysis (HRMA). (C) Three mutant alleles, gdf3zy51, gdf3zy52, gdf3z53, had 8, 1 and 6 bp deletions respectively. (D–G) Morphological phenotypes of gdf3 mutants at 24 hpf. (D) Wild-type (WT) and (E) zygotic (Z) gdf3 mutants were phenotypically indistinguishable. (F) Maternal-zygotic (MZ) gdf3 mutants lacked notochord, pharyngeal endoderm and had reduced anterior neural tissues. (G) MZgdf3 mutants were completely rescued by injection of 100 pg gdf3 RNA at the one-cell stage. (H–W) WISH analysis of gene expression in WT (columns H-T and J-V) and MZgdf3 (columns I-U and K-W) mutants at 24 hpf. (H, I) ta (ntl) expression in the notochord was absent from MZgdf3 although tailbud expression was maintained. (J, K) tbx16 (spt) expression in spinal cord neurons was absent in MZgdf3 while tailbud expression is unaffected. (L, M) Trunk and tail somites expressing myod1 were reduced in number and altered in shape in MZgdf3. (N, O) Expression of hand2 in the heart, pharyngeal arch mesoderm and pectoral fin buds is absent in MZgdf3. (P–S) Ventral neural tissues and pharyngeal endoderm expressing foxa2 (axial) were absent in MZgdf3. Patterns of expression of otx2 in the forebrain and midbrain (T, U), and erg2b (krox20) in hindbrain rhobomeres 3 and 5 (V, W) were altered in MZgdf3 mutants. All embryos (D–W) are shown in lateral view with rostral to the left except N, O, R, S which are dorsal views in the same orientation. Each panel is a representative image from at least 15 embryos.

Gene Expression Details
Gene Antibody Fish Conditions Stage Qualifier Anatomy Assay
egr2b AB standard conditions Prim-5 rhombomere 3 ISH
Prim-5 rhombomere 5 ISH
gdf3zy51/zy51(AB) standard conditions Prim-5 rhombomere 3 ISH
Prim-5 rhombomere 5 ISH
gdf3zy52/zy52(AB) standard conditions Prim-5 rhombomere 3 ISH
Prim-5 rhombomere 5 ISH
foxa2 AB standard conditions Prim-5 brain ventro-medial region ISH
Prim-5 floor plate spinal cord region ISH
Prim-5 pharyngeal endoderm ISH
gdf3zy51/zy51(AB) standard conditions Prim-5 Not Detected brain ventro-medial region ISH
Prim-5 Not Detected floor plate spinal cord region ISH
Prim-5 Not Detected pharyngeal endoderm ISH
gdf3zy52/zy52(AB) standard conditions Prim-5 Not Detected brain ventro-medial region ISH
Prim-5 Not Detected floor plate spinal cord region ISH
Prim-5 Not Detected pharyngeal endoderm ISH
hand2 AB standard conditions Prim-5 heart tube ISH
Prim-5 pectoral fin bud ISH
Prim-5 pharyngeal mesoderm ISH
gdf3zy51/zy51(AB) standard conditions Prim-5 Not Detected heart tube ISH
Prim-5 Not Detected pectoral fin bud ISH
Prim-5 Not Detected pharyngeal mesoderm ISH
gdf3zy52/zy52(AB) standard conditions Prim-5 Not Detected heart tube ISH
Prim-5 Not Detected pectoral fin bud ISH
Prim-5 Not Detected pharyngeal mesoderm ISH
myod1 AB standard conditions Prim-5 somite ISH
gdf3zy51/zy51(AB) standard conditions Prim-5 somite ISH
gdf3zy52/zy52(AB) standard conditions Prim-5 somite ISH
otx2b AB standard conditions Prim-5 forebrain ISH
Prim-5 midbrain ISH
gdf3zy51/zy51(AB) standard conditions Prim-5 forebrain ISH
Prim-5 midbrain ISH
gdf3zy52/zy52(AB) standard conditions Prim-5 forebrain ISH
Prim-5 midbrain ISH
tbx16 AB standard conditions Prim-5 spinal cord neuron ISH
Prim-5 tail bud ISH
gdf3zy51/zy51(AB) standard conditions Prim-5 Not Detected spinal cord neuron ISH
Prim-5 tail bud ISH
gdf3zy52/zy52(AB) standard conditions Prim-5 Not Detected spinal cord neuron ISH
Prim-5 tail bud ISH
tbxta AB standard conditions Prim-5 notochord ISH
Prim-5 tail bud ISH
gdf3zy51/zy51(AB) standard conditions Prim-5 Not Detected notochord ISH
Prim-5 tail bud ISH
gdf3zy52/zy52(AB) standard conditions Prim-5 Not Detected notochord ISH
Prim-5 tail bud ISH
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
gdf3zy51/zy51(AB) standard conditions Prim-5 brain ventro-medial region foxa2 expression absent, abnormal
Prim-5 central nervous system anterior region malformed, abnormal
Prim-5 eye fused with eye, abnormal
Prim-5 floor plate spinal cord region foxa2 expression absent, abnormal
Prim-5 heart tube hand2 expression absent, abnormal
Prim-5 notochord tbxta expression absent, abnormal
Prim-5 notochord absent, abnormal
Prim-5 pectoral fin bud hand2 expression absent, abnormal
Prim-5 pharyngeal endoderm foxa2 expression absent, abnormal
Prim-5 pharyngeal endoderm absent, abnormal
Prim-5 pharyngeal mesoderm hand2 expression absent, abnormal
Prim-5 somite decreased size, abnormal
Prim-5 somite myod1 expression spatial pattern, abnormal
Prim-5 somite U-shaped, abnormal
Prim-5 spinal cord neuron tbx16 expression absent, abnormal
gdf3zy51/zy51(AB) standard conditions Prim-5 whole organism morphology, normal
gdf3zy52/zy52(AB) standard conditions Prim-5 brain ventro-medial region foxa2 expression absent, abnormal
Prim-5 central nervous system anterior region malformed, abnormal
Prim-5 eye fused with eye, abnormal
Prim-5 floor plate spinal cord region foxa2 expression absent, abnormal
Prim-5 heart tube hand2 expression absent, abnormal
Prim-5 notochord absent, abnormal
Prim-5 notochord tbxta expression absent, abnormal
Prim-5 pectoral fin bud hand2 expression absent, abnormal
Prim-5 pharyngeal endoderm foxa2 expression absent, abnormal
Prim-5 pharyngeal endoderm absent, abnormal
Prim-5 pharyngeal mesoderm hand2 expression absent, abnormal
Prim-5 somite decreased size, abnormal
Prim-5 somite myod1 expression spatial pattern, abnormal
Prim-5 somite U-shaped, abnormal
Prim-5 spinal cord neuron tbx16 expression absent, abnormal
gdf3zy52/zy52(AB) standard conditions Prim-5 whole organism morphology, normal
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