header logo image header logo text
Downloads Login
Research
General Information
ZIRC
ZFIN ID: ZDB-FIG-180123-12
Wei et al., 2017 - A GEF activity-independent function for nuclear Net1 in Nodal/Smad2 signal transduction and mesendoderm formation. Journal of Cell Science   130(18):3072-3082 Full text @ J. Cell Sci.
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Fish:
Knockdown Reagent:
Anatomical Terms:
Stage Range: Shield to Prim-25
PHENOTYPE:
Fish:
Knockdown Reagent:
Observed In:
Stage Range: Shield to Prim-25

Fig. 1

net1 is crucial for mesendoderm formation in zebrafish embryos. (A) Visualization of net1 expression in zebrafish embryos at shield and 75% epiboly (ep) stages using whole-mount in situ hybridization. Shield stage, lateral views with dorsal to the right; 75% epiboly stage, dorsal views with animal pole to the top. (B–F) The expression of mesendoderm marker genes and mesendoderm derivative marker genes in cMO- and net1 MO1-injected embryos at the indicated stages. (B–D) Shield stage, animal views with dorsal to the right; 75% epiboly stage, dorsal views with animal pole to the top. (E,F) Bud stage, and 24 and 36 hpf. All images are shown in dorsal views with anterior to the top. (G) Wild-type embryos were injected with 50 pg ΔN-tcf3 mRNA alone or co-injected with 3 pg tar* mRNA and 4 ng net1 MO1, and then collected at the shield and 75% epiboly stages for whole-mount in situ hybridization with a gsc or sox17 probe. (H) Overexpression of net1 rescues mesendoderm induction defects in net1 morphants. Wild-type embryos were injected with cMO or net1 3Mix (4 ng net1 MO1, 200 pg Flag-net1 mRNA and 1 ng AS-Flag-photo-MO), and then a subset of net1 3Mix-injected embryos were exposed to UV light at the sphere stage. The expression of gsc and sox17 was examined by in situ hybridization at shield and 75% epiboly stages, respectively. In B–H, the percentages (mean±s.d.) of the affected embryos are shown as calculated from three independent biological repeats with ∼15–30 embryos in each group. UIC, uninjected control.

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
foxa1 TU control Prim-5 endoderm ISH
TU + MO1-net1 standard conditions Prim-5 endoderm ISH
gsc TU control Shield presumptive mesoderm ISH
75%-epiboly mesoderm ISH
TU + MO1-net1 standard conditions Shield presumptive mesoderm ISH
75%-epiboly mesoderm ISH
hhex TU control Prim-5 liver primordium ISH
Prim-5 pancreas primordium ISH
Prim-25 liver ISH
Prim-25 pancreatic bud ISH
TU + MO1-net1 standard conditions Prim-5 liver primordium ISH
Prim-5 pancreas primordium ISH
Prim-25 liver ISH
Prim-25 pancreatic bud ISH
net1 TU standard conditions Shield shield ISH
75%-epiboly axial mesoderm ISH
nkx2.3 TU control Prim-25 pharyngeal pouch ISH
TU + MO1-net1 standard conditions Prim-25 pharyngeal pouch ISH
shha TU control Bud notochord ISH
TU + MO1-net1 standard conditions Bud notochord ISH
sox17 TU control Shield presumptive endoderm ISH
75%-epiboly endoderm ISH
TU + MO1-net1 standard conditions Shield presumptive endoderm ISH
75%-epiboly endoderm ISH
sox32 TU control Shield presumptive endoderm ISH
75%-epiboly endoderm ISH
TU + MO1-net1 standard conditions Shield presumptive endoderm ISH
75%-epiboly endoderm ISH
tbxta TU control Shield presumptive mesoderm ISH
75%-epiboly mesoderm ISH
Bud notochord ISH
TU + MO1-net1 standard conditions Shield presumptive mesoderm ISH
75%-epiboly mesoderm ISH
Bud notochord ISH
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
TU + MO1-net1 standard conditions Shield presumptive endoderm sox32 expression decreased amount, abnormal
Shield presumptive endoderm sox17 expression decreased amount, abnormal
Shield presumptive mesoderm gsc expression decreased amount, abnormal
Shield presumptive mesoderm tbxta expression decreased amount, abnormal
75%-epiboly endoderm sox17 expression decreased amount, abnormal
75%-epiboly endoderm sox32 expression decreased amount, abnormal
75%-epiboly mesoderm tbxta expression decreased amount, abnormal
75%-epiboly mesoderm gsc expression decreased amount, abnormal
Bud notochord shha expression decreased amount, abnormal
Bud notochord tbxta expression decreased amount, abnormal
Bud notochord tbxta expression spatial pattern, abnormal
Bud notochord shha expression spatial pattern, abnormal
Prim-5 endoderm foxa1 expression decreased amount, abnormal
Prim-5 liver primordium hhex expression decreased amount, abnormal
Prim-5 pancreas primordium hhex expression decreased amount, abnormal
Prim-25 liver hhex expression decreased amount, abnormal
Prim-25 pancreatic bud hhex expression decreased amount, abnormal
Prim-25 pharyngeal pouch nkx2.3 expression decreased amount, abnormal
Acknowledgments:
ZFIN wishes to thank the journal Journal of Cell Science for permission to reproduce figures from this article. Please note that this material may be protected by copyright. Full text @ J. Cell Sci.