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Experiment details for tbxt

Kam RK et al. (2013) Assay

Dhrs3 protein attenuates retinoic acid signaling and is required for early embryonic patterning.

Gene Clone Species Stages Anatomy
tbxt.S laevis NF stage 10.25 marginal zone , ventral marginal zone , ventro-lateral marginal zone , dorso-lateral marginal zone , dorsal marginal zone , [+]

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  FIGURE 6. Expression patterns of mesoderm and organizer marker genes in Dhrs3 morphants and embryos treated with 1 μM atRA during gastrulation. Embryos were injected at the two-cell stage with either 30 ng of Dhrs3 MO or 30 ng of Dhrs3 MO plus 1 ng of dhrs3 mRNA or treated with 1 μM of atRA at gastrulation stages. Whole mount in situ hybridization was used to visualize mesoderm marker gene brachyury (bra) (A1�A4), organizer marker genes lhx1 (B1�B4), and goosecoid (gsc) (C1�C4). Expression of bra was disrupted in Dhrs3 morphants (A2, arrow) and in embryos treated with atRA (A4, arrow). Injection of 1 ng of dhrs3 mRNA rescued the disruption (A3). The expression intensity of lhx1 was elevated by Dhrs3 knockdown (B2) and atRA treatment (B4). Compared with the control embryos (C1), the expression domain of gsc was closer to the blastopore in Dhrs3 morphants (C2) and was abolished by atRA treatment (C4). Dotted line outlined the blastopore.

Gene Clone Species Stages Anatomy
tbxt.S laevis NF stage 13 axial mesoderm , posterior

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  FIGURE 7. Defective convergent extension movement induced by knockdown of Dhrs3. Expression of mesoderm and notochord marker genes brachyury (bra), not, and goosecoid (gsc), and CE marker gene papc in control embryos (A-D), in response to dhrs3 knockdown (E-H), dhrs3 overexpression (I-L), or exogenous atRA (M-P). The expression domain of bra and not was shortened by Dhrs3 knockdown (E and F) and embryos treated with atRA (M and N). A similar but less severe shortening of bra expression domain was observed in embryos overexpressing dhrs3 (I). The not expression domain remained in normal shape, but the signal was less intensified in embryos overexpressing dhrs3 (J). The expression of gsc was suppressed by Dhrs3 knockdown (G) and atRA treatment (O) but was unaffected by dhrs3 overexpression (K). papc expression domain was more diffused in Dhrs3 morphants (H) and atRA treated embryos (P) when compared with control embryos (D). Percentages of embryos showing the phenotypes were marked in the figure. All embryos are dorsal view.