ZFIN ID: ZDB-FIG-060307-9
Rentzsch et al., 2006 - Crossveinless 2 is an essential positive feedback regulator of Bmp signaling during zebrafish gastrulation. Development (Cambridge, England)   133(5):801-811 Full text @ Development
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Fish:
Knockdown Reagent:
Anatomical Terms:
Stage Range: 75%-epiboly to 10-13 somites

Fig. 2 cvl2 is required for ventral specification and Bmp signaling. (A) Western blot; translation of myc-tagged cvl2 mRNA is suppressed by cvl2 ATG-MO. (B) Western blots, revealing strongly reduced pSmad1/5 levels in embryos of 80% epiboly stage after injection of cvl2 MO1 (upper panel) or co-injection of cvl2 MO1+2 (lower panel). (C,D) Whole-mount immunostaining, lateral view at 80% epiboly; cvl2 MO1 leads to a reduction of ventral α-pSmad1/5 immunoreactivity. (E,F) p-Smad1/5 intensity along dorsoventral axis of cvl2 morphant (gray in F) and control sibling (black in F) at 80% epiboly stage. Signal intensities were measured from vegetal views, along a yellow line indicated in E for one of the investigated controls (n=7). (G-X) Whole-mount in situ hybridization with probes indicated in lower left corners. (G-N) Eighty percent epiboly stage. (G,H,K-N) Lateral views; (I,J) vegetal views, dorsal rightwards. Ratios of affected embryos were: eve1, 33/39; cvl2, 66/71; gata2, 31/32; otx2, 45/53. (J) Only the ventral expression of cvl2 is reduced in the morphant, while expression in the prechordal plate is normal. (O,P) Ten-somite stage, view of tailbud, anterior leftwards; expression domain of the blood precursor marker gata1 is reduced in cvl2 morphant (29/37 embryos). (Q-X) Five-somite stage, anterior leftwards; the krox20 expression domains are ventrally expanded in cvl2 morphant (R, 53/68 embryos), but rescued to near wild-type condition in embryo co-injected with cvl2 MO and Drosophila cvl2 mRNA (T, 46/47 embryos). (U-X) Low doses of cvl2 MO1 enhance the mild dorsalization of tolloid/minifin mutants (40/43 in cross of two mfn homozygotes, 22/97 in cross of two mfn heterozygotes), while having no or a much weaker effect in wild-type embryos injected in parallel (0/33). Control embryos are injected with a mismatch cvl2 morpholino. Abbreviations: Coom, Coomassie Blue staining of gel loaded with same amounts of protein; PonRed, Ponceau Red staining of blots after immunoreaction.

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
bmper WT standard conditions 75%-epiboly blastoderm ISH
75%-epiboly prechordal plate ISH
WT + MO1-bmper standard conditions 75%-epiboly prechordal plate ISH
egr2b WT standard conditions 5-9 somites presumptive rhombomere 3 ISH
5-9 somites presumptive rhombomere 5 ISH
WT + MO1-bmper standard conditions 5-9 somites presumptive rhombomere 3 ISH
5-9 somites presumptive rhombomere 5 ISH
tll1tc263a/tc263a(TU) standard conditions 5-9 somites presumptive rhombomere 3 ISH
5-9 somites presumptive rhombomere 5 ISH
eve1 WT standard conditions 75%-epiboly ventral mesoderm ISH
WT + MO1-bmper standard conditions 75%-epiboly ventral mesoderm ISH
gata1a WT standard conditions 10-13 somites lateral mesoderm ISH
WT + MO1-bmper standard conditions 10-13 somites lateral mesoderm ISH
gata2a WT standard conditions 75%-epiboly ectoderm ISH
WT + MO1-bmper standard conditions 75%-epiboly ectoderm ISH
gsc WT standard conditions 75%-epiboly blastoderm ISH
WT + MO1-bmper standard conditions 75%-epiboly blastoderm ISH
otx2b WT standard conditions 75%-epiboly neuroectoderm ISH
WT + MO1-bmper standard conditions 75%-epiboly neuroectoderm ISH
Antibody Labeling Details No data available
Phenotype Details No data available
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