ZFIN ID: ZDB-FIG-050120-6
Little et al., 2004 - Twisted gastrulation promotes BMP signaling in zebrafish dorsal-ventral axial patterning. Development (Cambridge, England)   131(23):5825-5835 Full text @ Development
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Fish:
Knockdown Reagent:
Anatomical Terms:
Stage Range: 75%-epiboly to 5-9 somites

Fig. 2 High level tsg1 knockdown dorsalizes the embryo. Uninjected embryos (A-E,K-M) compared with siblings injected with 25 ng MO5 (F-J,N,P,Q) or 32 ng MO1 (O). (A-J) Whole-mount in situ hybridization at 80% epiboly (mid-gastrulation); animal pole views, dorsal towards the right. Reduced ventral domain of eve1 (n=32/54), dlx3 (n=25/41) and gata2 (n=28/50) expression (A-C,F-H) and expanded foxb1.2 (n=34/46) and chordin (chd) (n=16/30) expression in dorsal regions (D,E,I,J) in injected compared with uninjected embryos (delineated by arrowheads) was observed. gsc expression in the dorsal midline prechordal plate mesoderm is not affected (A,F), similar to results in other dorsalized BMP signaling pathway mutants in zebrafish (Mullins et al., 1996; Nguyen et al., 1998). (K-Q) During somitogenesis, injected embryos display dorsalized characteristics similar to class 3 (N) and class 4 (O) dorsalized mutants, as assessed by the lateral extent of somites in live embryos (arrowheads in K,N,O), by whole-mount in situ hybridization with myod in anterior somitic mesoderm (M,P,Q), and by expansion of pax2.1 (arrowhead) and krox20 (bracket) expression in the MHB and rhombomeres 3 and 5, respectively (L,Q). The embryos in N,P display a class 3 or moderate dorsalization, whereas those in O,Q exhibit a greater expansion of the somitic mesoderm and neural tissue, similar to a class 4 dorsalization. (L) Dorsoanterior view; (M) a more posterior view of the same uninjected embryo. (R) Anti-FLAG western blot on lysates of embryos injected with FLAG-tagged tsg1 alone or with 32 ng MO1 or mismatch MO1. Bars to the left of the blot represent the positions of protein standards with molecular weights of 50, 37 and 25 kDa.

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
chrd WT standard conditions 75%-epiboly unspecified ISH
WT + MO3-twsg1b standard conditions 75%-epiboly unspecified ISH
dlx3b WT standard conditions 75%-epiboly unspecified ISH
WT + MO3-twsg1b standard conditions 75%-epiboly unspecified ISH
egr2b WT standard conditions 5-9 somites presumptive rhombomere 3 ISH
5-9 somites presumptive rhombomere 5 ISH
WT + MO3-twsg1b standard conditions 5-9 somites presumptive rhombomere 3 ISH
5-9 somites presumptive rhombomere 5 ISH
eve1 WT standard conditions 75%-epiboly unspecified ISH
WT + MO3-twsg1b standard conditions 75%-epiboly unspecified ISH
foxb1a WT standard conditions 75%-epiboly unspecified ISH
WT + MO3-twsg1b standard conditions 75%-epiboly unspecified ISH
gata2a WT standard conditions 75%-epiboly unspecified ISH
WT + MO3-twsg1b standard conditions 75%-epiboly unspecified ISH
gsc WT standard conditions 75%-epiboly unspecified ISH
WT + MO3-twsg1b standard conditions 75%-epiboly unspecified ISH
myod1 WT standard conditions 5-9 somites mesoderm ISH
WT + MO3-twsg1b standard conditions 5-9 somites mesoderm ISH
pax2a WT standard conditions 5-9 somites midbrain hindbrain boundary neural rod ISH
WT + MO3-twsg1b standard conditions 5-9 somites midbrain hindbrain boundary neural rod ISH
Antibody Labeling Details No data available
Phenotype Details No data available
Acknowledgments:
This image is the copyrighted work of the attributed author or publisher, and ZFIN has permission only to display this image to its users. Additional permissions should be obtained from the applicable author or publisher of the image. Full text @ Development