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ZFIN ID: ZDB-FIG-180503-5
Pelliccia et al., 2017 - Gdf3 is required for robust Nodal signaling during germ layer formation and left-right patterning. eLIFE   6 Full text @ Elife
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Antibody:
Fish:
Anatomical Terms:
Stage: Shield
PHENOTYPE:
Fish:
Knockdown Reagents:
Observed In:
Stage Range: Shield to Prim-5

Fig. 3

Loss of Gdf3 leads to expansion of Bmp signaling.

(A) Knockdown of nodal related 1 (ndr1) and nodal related 2 (ndr2) in wild-type (WT) embryos causes complete loss of head and trunk mesendoderm and defects in tail patterning. (B) ndr1:ndr2 knockdown in MZgdf3 embryos causes loss of residual tail patterning, producing embryos that more closely resemble those completely lacking Nodal signaling (A). (C-E) Overexpression of 1 pg or 10 pg ndr1 led to increased expression of the Nodal target gene goosecoid (gsc) in WT embryos. (F-H) Overexpression of 10 pg ndr1, but not 1 pg, produced widespread expression of gsc in MZgdf3 mutants. (I) Immunostaining of phosphorylated Smad1/5/8 (green) counterstained with DAPI (blue). Ventral pSmad levels were increased in MZgdf3 mutants. (J) Plotting the normalized pSmad1/5/8 intensity over position on the ventral-dorsal axis confirms that MZgdf3 embryos (n = 14) have increased BMP signaling in the ventral region compared to WT (n = 13). (K-M) RNA in situ hybridization of eve1 showed expansion of expression in the ventral region of MZgdf3 embryos (K) compared to WT (L). (M) Quantitative comparison of WT (n = 14) and MZgdf3 (n = 12) embryos confirmed expansion of the ventral eve1 expression domain in MZgdf3 embryos. (N-Q) RNA in situ hybridization of chordin (chd) revealed its expression is reduced in MZgdf3 embryos. All views are animal unless otherwise indicated. In M, p-value obtained by Student’s t-test (two-sided, homoscedastic). Error bars in J and M, standard error of the mean. Matlab code and data are available as Figure 3—source code 1, 2, and 3 and Figure 3—source data 1 and 2.

Gene Expression Details
Gene Antibody Fish Conditions Stage Qualifier Anatomy Assay
chrd WT standard conditions Shield whole organism dorsal region ISH
gdf3pr05/pr05 standard conditions Shield whole organism dorsal region ISH
eve1 WT standard conditions Shield whole organism ventral region ISH
gdf3pr05/pr05 standard conditions Shield whole organism ventral region ISH
gsc WT standard conditions Shield whole organism dorsal region ISH
gdf3pr05/pr05 standard conditions Shield Not Detected whole organism dorsal region ISH
Antibody Labeling Details
Antibody Assay Fish Conditions Stage Anatomy
Ab1-smad IHC WT standard conditions Shield whole organism ventral region
IHC gdf3pr05/pr05 standard conditions Shield whole organism ventral region
Phenotype Details
Fish Conditions Stage Phenotype
WT + MO1-ndr2 + MO4-ndr1 standard conditions Prim-5 caudal fin morphology, abnormal
Prim-5 endoderm formation decreased occurrence, abnormal
Prim-5 mesoderm formation decreased occurrence, abnormal
gdf3pr05/pr05 standard conditions Shield whole organism dorsal region gsc expression absent, abnormal
Shield whole organism dorsal region chrd expression decreased amount, abnormal
Shield whole organism dorsal region chrd expression decreased distribution, abnormal
Shield whole organism ventral region ab1-smad labeling increased amount, abnormal
Shield whole organism ventral region eve1 expression increased amount, abnormal
Shield whole organism ventral region eve1 expression increased distribution, abnormal
Shield whole organism ventral region ab1-smad labeling increased distribution, abnormal
gdf3pr05/pr05 + MO1-ndr2 + MO4-ndr1 standard conditions Prim-5 caudal fin morphology, exacerbated
Prim-5 endoderm formation decreased occurrence, abnormal
Prim-5 mesoderm formation decreased occurrence, 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