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ZFIN ID: ZDB-FIG-131211-61
Arrington et al., 2013 - Sdc2 and Tbx16 regulate Fgf2-dependent epithelial cell morphogenesis in the ciliated organ of asymmetry. Development (Cambridge, England)   140(19):4102-4109 Full text @ Development
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Antibodies:
Fish:
Knockdown Reagent:
Anatomical Terms:
Stage Range: 75%-epiboly to 5-9 somites
PHENOTYPE:
Fish:
Knockdown Reagents:
Observed In:
Stage Range: 75%-epiboly to 14-19 somites

Fig. 5 Sdc2 controls DFC/KV-specific expression of Tbx16, upstream of both Fgf2-dependent KV morphogenesis and Fgf2-independent cilia length regulation. (A-F) KV morphogenesis and cilia length defects in spadetail (spt; tbx16) mutants. (A,C) Normal KV morphology (A) and cilia length (C) in wild type or spt heterozygotes. (B,D) spt mutants had defective KV morphogenesis (B) and cilia length (D), similar to sdc2 morphants. (E,F) Quantification of (E) cilia length and (F) cilia number (spt+/?: eight embryos, 361 cilia; spt-/-: nine embryos, 242 cilia) indicated that both are decreased in spt mutants (*P<0.05, **P<0.001). Error bars indicate s.e.m. (G-L) Sdc2 regulates Tbx16 expression in DFCs/KV. (G,H) Green arrowheads indicate (G) tbx16 RNA and (H) Tbx16 protein expression in DFCs in wild-type embryos. (I,J) Red arrowheads indicate the position of DFCs lacking (I) tbx16 mRNA and (J) Tbx16 protein expression in sdc2 morphants. By contrast, tbx16 RNA and Tbx16 protein expression in adjacent non-DFC domains was normal, revealing a domain-specific regulation of Tbx16 by Sdc2. (K) fgf2 was downregulated in spt mutants (compare with wild type in Fig. 4B). (L) Synergy between Sdc2, Tbx16 and Fgf2 was tested by injection of subeffective amounts of each MO individually (sdc2 SB2 MO, n=431; tbx16 MO, n=396; fgf2 MO, n=475) compared with co-injections of pairs of subthreshold MOs. Co-injection of sdc2 SB2 and tbx16 MOs (n=251) and co-injection of tbx16 and fgf2 MOs (n=111) resulted in significantly affected spaw expression (P<0.001) compared with single injections of subthreshold MOs. P-values by Student’s t-test.

Gene Expression Details
Gene Antibody Fish Conditions Stage Qualifier Anatomy Assay
fgf2 tbx16b104/b104 standard conditions Bud Not Detected forerunner cell group ISH
tbx16 AB standard conditions 75%-epiboly forerunner cell group ISH
75%-epiboly forerunner cell group IHC
75%-epiboly margin ISH
75%-epiboly margin IHC
75%-epiboly presumptive paraxial mesoderm IHC
75%-epiboly presumptive paraxial mesoderm ISH
AB + MO5-sdc2 standard conditions 75%-epiboly Not Detected forerunner cell group IHC
75%-epiboly Not Detected forerunner cell group ISH
75%-epiboly margin ISH
75%-epiboly margin IHC
75%-epiboly presumptive paraxial mesoderm ISH
75%-epiboly presumptive paraxial mesoderm IHC
Antibody Labeling Details
Antibody Assay Fish Conditions Stage Anatomy
Ab1-prkcz IHC AB standard conditions 5-9 somites Kupffer's vesicle apical plasma membrane
IHC tbx16b104/b104 standard conditions 5-9 somites Kupffer's vesicle apical plasma membrane
Ab1-tuba IHC AB standard conditions 5-9 somites Kupffer's vesicle motile cilium
IHC tbx16b104/b104 standard conditions 5-9 somites Kupffer's vesicle motile cilium
Phenotype Details
Fish Conditions Stage Phenotype
AB + MO1-fgf2 + MO3-tbx16 standard conditions 14-19 somites determination of left/right asymmetry in lateral mesoderm process quality, abnormal
AB + MO3-tbx16 + MO5-sdc2 standard conditions 14-19 somites determination of left/right asymmetry in lateral mesoderm process quality, abnormal
AB + MO5-sdc2 standard conditions 75%-epiboly forerunner cell group physical object quality, abnormal
tbx16b104/b104 standard conditions Bud fibroblast growth factor receptor signaling pathway process quality, abnormal
Bud Kupffer's vesicle physical object quality, abnormal
Bud Kupffer's vesicle development process quality, abnormal
5-9 somites Kupffer's vesicle decreased size, abnormal
5-9 somites Kupffer's vesicle malformed, abnormal
5-9 somites Kupffer's vesicle development process quality, abnormal
5-9 somites Kupffer's vesicle motile cilium decreased amount, abnormal
5-9 somites Kupffer's vesicle motile cilium decreased length, abnormal
Acknowledgments:
ZFIN wishes to thank the journal Development (Cambridge, England) for permission to reproduce figures from this article. Please note that this material may be protected by copyright. Full text @ Development