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ZFIN ID: ZDB-FIG-171227-7
Shao et al., 2017 - Vegetally localised Vrtn functions as a novel repressor to modulate bmp2b transcription during dorsoventral patterning in zebrafish.. Development (Cambridge, England)   144:3361-3374 Full text @ Development
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Fish:
Condition:
Anatomical Terms:
Stage Range: Sphere to 5-9 somites
PHENOTYPE:
Fish:
Condition:
Observed In:
Stage Range: 50%-epiboly to Prim-5

Fig. 5

The dorsalising effect of VAb@2cell is suppressed in MZvrtn mutants. (A) Targeted mutation of the vrtn gene. The Cas9/gRNA targeting sequence and the PAM region (red) are shown, and the mutated allele with the predicted peptide is indicated. (B) Representative images of indicated embryos at 27 hpf, showing smaller eyes and reduced head size in vrtn mutants. (C) Decreased rx3 and six3b expression domains in vrtn mutants at the bud stage. (D) Expression patterns of six3b and myod1 in flat-mounted embryos with equal somite numbers. (E,F) Analysis by scatter plots of head length and six3b expression area at the six-somite stage. The red broken lines in the insets define the parameters of measurement. Horizontal lines represent the mean±s.d. from three batches of embryos with the total number indicated. (G-J) Analyses by in situ hybridisation and qRT-PCR of the expression of gsc, dha/boz, bmp2b and otx2 in MZvrtn mutants. The expression of gsc and dha/boz remains unchanged at sphere stage (animal pole views), the expression of bmp2b is enhanced at 50% epiboly, and the expression of otx2 is reduced at 80% epiboly (lateral views with dorsal on the right). The relative expression level in wild-type embryos is normalised as 1; data are mean±s.d. from three independent experiments. (K) MZvrtn embryos are ventralised following VAb@2cell. (L) Statistical analysis of the ventralising effect. Numbers at the top represent total embryos from three independent experiments. (M,N) Analyses by in situ hybridisation and qRT-PCR of bmp2b expression at the shield stage in wild-type and MZvrtn embryos after VAb@2cell. The embryos are shown as lateral views. The relative bmp2b level in VAb@2cell wild-type embryos is set as 1; data are mean±s.d. from six independent experiments. Scale bars: 250 µm.

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
bmp2b WT standard conditions 50%-epiboly anatomical structure ISH
WT ablation: yolk Shield margin ISH
vrtnsdu1/sdu1 standard conditions 50%-epiboly anatomical structure ISH
50%-epiboly whole organism RTPCR
vrtnsdu1/sdu1 ablation: yolk Shield margin RTPCR
Shield margin ISH
dharma WT standard conditions Sphere anatomical structure ISH
vrtnsdu1/sdu1 standard conditions Sphere anatomical structure ISH
Sphere whole organism RTPCR
gsc WT standard conditions Sphere anatomical structure ISH
vrtnsdu1/sdu1 standard conditions Sphere anatomical structure ISH
Sphere whole organism RTPCR
myod1 WT standard conditions 5-9 somites somite ISH
vrtnsdu1/sdu1 standard conditions 5-9 somites somite ISH
vrtnsdu1 standard conditions 5-9 somites somite ISH
otx2b WT standard conditions 75%-epiboly anatomical structure ISH
vrtnsdu1/sdu1 standard conditions 75%-epiboly anatomical structure ISH
75%-epiboly whole organism RTPCR
rx3 WT standard conditions Bud optic primordium ISH
vrtnsdu1/sdu1 standard conditions Bud optic primordium ISH
vrtnsdu1 standard conditions Bud optic primordium ISH
six3b WT standard conditions Bud forebrain neural plate ISH
5-9 somites forebrain neural rod ISH
vrtnsdu1/sdu1 standard conditions Bud forebrain neural plate ISH
5-9 somites forebrain neural rod ISH
vrtnsdu1 standard conditions Bud forebrain neural plate ISH
5-9 somites forebrain neural rod ISH
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
WT ablation: yolk 5-9 somites whole organism wholly dorsalized, abnormal
Prim-5 whole organism wholly dorsalized, abnormal
vrtnsdu1/sdu1 standard conditions 50%-epiboly anatomical structure bmp2b expression increased amount, abnormal
50%-epiboly whole organism bmp2b expression increased amount, abnormal
75%-epiboly anatomical structure otx2b expression decreased amount, abnormal
75%-epiboly whole organism otx2b expression decreased amount, abnormal
Bud forebrain neural plate six3b expression decreased distribution, abnormal
Bud optic primordium rx3 expression decreased distribution, abnormal
5-9 somites forebrain neural rod six3b expression decreased distribution, abnormal
5-9 somites head decreased length, abnormal
Prim-5 head decreased size, abnormal
Prim-5 whole organism wholly ventralized, abnormal
vrtnsdu1/sdu1 ablation: yolk Shield margin bmp2b expression increased amount, abnormal
Shield margin bmp2b expression increased amount, abnormal
5-9 somites whole organism wholly ventralized, abnormal
Prim-5 whole organism wholly ventralized, abnormal
vrtnsdu1 standard conditions Bud forebrain neural plate six3b expression decreased distribution, abnormal
Bud optic primordium rx3 expression decreased distribution, abnormal
5-9 somites forebrain neural rod six3b expression decreased distribution, abnormal
5-9 somites head decreased length, abnormal
Prim-5 head decreased size, abnormal
Prim-5 whole organism wholly ventralized, 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