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Experiment details for hhex

Fletcher G et al. (2006) Assay

A role for GATA factors in Xenopus gastrulation movements.

Gene Clone Species Stages Anatomy
hhex.L laevis NF stage 10.5 endomesoderm

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  Fig. 1. GATA4 and GATA6 are expressed in migratory leading edge mesendoderm throughout gastrulation. (A–L) In situ hybridisation on embryos just prior to (stage 9), during (stages 10.25 and 12) and at the end of (stage 13) gastrulation. Serial sections were hybridised with probes for GATA6, GATA4 and the mesodermal marker Xbra. (M–O) In situ hybridisation showing coexpression of GATA6 with cerberus and Xhex (markers of leading edge mesendoderm) during gastrulation. The vegetal pole is to the bottom and dorsal is to the right in all panels except J–L, where dorsal is towards the top and anterior is left. Arrows in (A) and (B) denote suprablastoporal endoderm, where punctate staining is visible in a few cells. Arrow in (J) denotes archenteron roof. All sections are sagittal and 10 μm.

Gene Clone Species Stages Anatomy
hhex.L laevis NF stage 11 endomesoderm

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  Fig. 3. Inhibition of GATA function disrupts gastrulation movements. (A) Induction of Sox17α by mdlGATA6 in animal caps is inhibited by myqGATA6EnR. Relative gene expression in animal caps at sib stage 12 measured by real-time RT-PCR. Fifty picograms of each RNA was injected into animal caps of one-cell embryos. This experiment was repeated twice giving similar results each time. (B) Transactivation of a GATA-driven luciferase reporter gene by endogenous factors in the marginal zone is inhibited by myqGATA6EnR. The marginal zone of each blastomere at the four-cell stage was injected with reporter DNA ±myqGATA6EnR mRNA. Luciferase activity is expressed as fold activation relative to background activity in uninjected embryos. This experiment was repeated twice giving similar results each time. (C and D) Inhibition of GATA function disrupts gastrulation movements and anterior development. (C) myqGATA6EnR-injected embryos display delayed gastrulation, poor head development and shortened anterior posterior axes. Fluorescent image shows nlsGFP expression in a myqGATA6EnR-injected embryo (see bright-field image above) and demonstrates accurate targeting of the marginal zone. (D) Pooled embryo phenotype counts from three independent experiments. Only predominant defects were scored to avoid multiple counting of the same specimen. The phenotype observed varied between embryo batches, however gastrulation was always delayed and disturbed. (E) Morphology and gene expression in myqGATA6EnR-injected embryos. Following injection of myqGATA6EnR or EnR (control) RNA into each blastomere at the four cell stage, embryos were cultured to stage 11, sagitally sectioned and hybridised with anti-sense probes for the indicated genes. Morphology and gene expression are normal in EnR-injected embryos, but are disrupted by myqGATA6EnR-injection.