ZFIN ID: ZDB-FIG-120522-15
Das et al., 2012 - Bmps and id2a act upstream of twist1 to restrict ectomesenchyme potential of the cranial neural crest. PLoS Genetics   8(5):e1002710 Full text @ PLoS Genet.
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Fish:
Knockdown Reagents:
Anatomical Terms:
Stage Range: 14-19 somites to Long-pec
PHENOTYPE:
Fish:
Knockdown Reagents:
Observed In:
Stage: Day 5

Fig. 1

Twist1 genes are required for ectomesenchyme specification in zebrafish.

(A–D) Whole mount in situ hybridizations of sox10 expression at 18 hpf show ectopic expression in the arches (numbered and second arch indicated by black arrow) of twist1a/1b-MO (n = 16/16) and sox10:Gal4VP16; UAS:dnTwist1b (n = 4/4) embryos compared to un-injected (n = 0/14) and sox10:Gal4VP16 only (n = 0/9) controls. White arrowheads indicate otic expression. (E–H) Whole mount in situs at 24 hpf show reduction of fli1a expression in the arch ectomesenchyme (numbered) of twist1a/1b-MO (n = 12/12) and sox10:Gal4VP16; UAS:dnTwist1b (n = 5/5) embryos compared to un-injected (n = 0/13) and sox10:Gal4VP16 only (n = 0/8) controls. Insets in E and F highlight arch ectomesenchyme which is reduced in twist1a/1b-MO embryos. Vascular expression of fli1a (red arrowheads) is unaffected. (I–L) Whole mount in situs at 18 hpf show a slight reduction of dlx2a in sox10:Gal4VP16; UAS:dnTwist1b embryos (n = 4/4) but not un-injected (n = 0/6), twist1a/1b-MO (n = 0/8), and sox10:Gal4VP16 only (n = 0/6) embryos. (M-P) Skeletal staining at 5 dpf shows severe loss of CNCC-derived head skeleton in twist1a/1b-MO embryos (n = 21/21) and primarily jaw reductions in sox10:Gal4VP16; UAS:dnTwist1b embryos (n = 9/9) compared to no defects in un-injected (n = 0/24) and sox10:Gal4VP16 only (n = 0/16) controls. Whereas only small remnants remain of the CNCC-derived skeleton (arrows), the mesoderm-derived otic capsule cartilage (arrowheads) and posterior neurocranium are less affected in twist1a/1b-MO embryos. (Q–T) In situs for dct expression at 28 hpf show normal melanophore precursors in un-injected (n = 14), twist1a/1b-MO (n = 12), sox10:Gal4VP16 only (n = 8), and sox10:Gal4VP16; UAS:dnTwist1b (n = 8) embryos. (U–X) In situs for xdh expression at 28 hpf show normal xanthophore precursors in un-injected (n = 7), twist1a/1b-MO (n = 5), sox10:Gal4VP16 only (n = 9), and sox10:Gal4VP16; UAS:dnTwist1b (n = 8) embryos. (Y-AB) In situs for foxd3 expression at 48 hpf reveal largely normal patterns of glia in un-injected (n = 9), twist1a/1b-MO (n = 8), sox10:Gal4VP16 only (n = 9), and sox10:Gal4VP16; UAS:dnTwist1b (n = 8) embryos. Scale bar = 50 μm.

Gene Expression Details
Gene Antibody Fish Conditions Stage Qualifier Anatomy Assay
aox5 WT standard conditions Prim-5 xanthoblast ISH
WT + MO1-twist1a + MO3-twist1b standard conditions Prim-5 xanthoblast ISH
el159Tg standard conditions Prim-5 xanthoblast ISH
el159Tg; el179Tg standard conditions Prim-5 xanthoblast ISH
dct WT standard conditions Prim-5 melanoblast ISH
Prim-5 melanocyte ISH
WT + MO1-twist1a + MO3-twist1b standard conditions Prim-5 melanoblast ISH
Prim-5 melanocyte ISH
el159Tg standard conditions Prim-5 melanoblast ISH
Prim-5 melanocyte ISH
el159Tg; el179Tg standard conditions Prim-5 melanoblast ISH
Prim-5 melanocyte ISH
dlx2a WT standard conditions 14-19 somites pharyngeal arch ISH
14-19 somites pharyngeal arch 1 ISH
14-19 somites pharyngeal arch 2 ISH
14-19 somites pharyngeal arch 3 ISH
WT + MO1-twist1a + MO3-twist1b standard conditions 14-19 somites pharyngeal arch ISH
14-19 somites pharyngeal arch 1 ISH
14-19 somites pharyngeal arch 2 ISH
14-19 somites pharyngeal arch 3 ISH
el159Tg standard conditions 14-19 somites pharyngeal arch ISH
14-19 somites pharyngeal arch 1 ISH
14-19 somites pharyngeal arch 2 ISH
14-19 somites pharyngeal arch 3 ISH
el159Tg; el179Tg standard conditions 14-19 somites pharyngeal arch ISH
14-19 somites pharyngeal arch 1 ISH
14-19 somites pharyngeal arch 2 ISH
14-19 somites pharyngeal arch 3 ISH
fli1 WT standard conditions Prim-5 pharyngeal arch ectomesenchyme ISH
Prim-5 pharyngeal arch 1 ISH
Prim-5 pharyngeal arch 2 ISH
Prim-5 pharyngeal arch 3 ISH
Prim-5 pharyngeal arch 4 ISH
Prim-5 vasculature ISH
WT + MO1-twist1a + MO3-twist1b standard conditions Prim-5 pharyngeal arch ectomesenchyme ISH
Prim-5 pharyngeal arch 1 ISH
Prim-5 pharyngeal arch 2 ISH
Prim-5 pharyngeal arch 3 ISH
Prim-5 pharyngeal arch 4 ISH
Prim-5 vasculature ISH
el159Tg standard conditions Prim-5 pharyngeal arch ectomesenchyme ISH
Prim-5 pharyngeal arch 1 ISH
Prim-5 pharyngeal arch 2 ISH
Prim-5 pharyngeal arch 3 ISH
Prim-5 pharyngeal arch 4 ISH
Prim-5 vasculature ISH
el159Tg; el179Tg standard conditions Prim-5 pharyngeal arch ectomesenchyme ISH
Prim-5 pharyngeal arch 1 ISH
Prim-5 pharyngeal arch 2 ISH
Prim-5 pharyngeal arch 3 ISH
Prim-5 pharyngeal arch 4 ISH
Prim-5 vasculature ISH
foxd3 WT standard conditions Long-pec glial cell ISH
WT + MO1-twist1a + MO3-twist1b standard conditions Long-pec glial cell ISH
el159Tg standard conditions Long-pec glial cell ISH
el159Tg; el179Tg standard conditions Long-pec glial cell ISH
sox10 WT standard conditions 14-19 somites otic vesicle ISH
14-19 somites Not Detected pharyngeal arch ISH
WT + MO1-twist1a + MO3-twist1b standard conditions 14-19 somites otic vesicle ISH
14-19 somites pharyngeal arch ISH
14-19 somites pharyngeal arch 1 ISH
14-19 somites pharyngeal arch 2 ISH
14-19 somites pharyngeal arch 3 ISH
el159Tg standard conditions 14-19 somites otic vesicle ISH
14-19 somites Not Detected pharyngeal arch ISH
el159Tg; el179Tg standard conditions 14-19 somites otic vesicle ISH
14-19 somites pharyngeal arch ISH
14-19 somites pharyngeal arch 1 ISH
14-19 somites pharyngeal arch 2 ISH
14-19 somites pharyngeal arch 3 ISH
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
WT + MO1-twist1a + MO3-twist1b standard conditions Day 5 mandibular arch skeleton hypoplastic, abnormal
Day 5 splanchnocranium hypoplastic, abnormal
el159Tg; el179Tg standard conditions Day 5 mandibular arch skeleton hypoplastic, abnormal
Day 5 splanchnocranium hypoplastic, abnormal
Acknowledgments:
ZFIN wishes to thank the journal PLoS Genetics for permission to reproduce figures from this article. Please note that this material may be protected by copyright. Full text @ PLoS Genet.