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First published online 19 January 2005
doi: 10.1242/dev.01647
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,
Institute of Genetics, University of Nottingham, Queen's Medical Centre, Nottingham NG7 2UH, UK
Author for correspondence (e-mail:
roger.patient{at}imm.ox.ac.uk)
Accepted 10 December 2004
The individual contributions of the three vertebrate GATA factors to
endoderm formation have been unclear. Here we detail the early expression of
GATA4, 5 and 6 in presumptive endoderm in Xenopus embryos and their
induction of endodermal markers in presumptive ectoderm. Induction of
HNF3ß by all three GATA factors was abolished when protein synthesis was
inhibited, showing that these inductions are indirect. In contrast, whereas
induction of Sox17
and HNF1ß by GATA4 and 5 was substantially
reduced when protein synthesis was inhibited, induction by GATA6 was minimally
affected, suggesting that GATA6 is a direct activator of these early
endodermal genes. GATA4 induced GATA6 expression in the same assay and
antisense morpholino oligonucleotides (MOs), designed to knock down
translation of GATA6, blocked induction of Sox17
and HNF1ß by
GATA4, suggesting that GATA4 induces these genes via GATA6 in this assay. All
three GATA factors were induced by activin, although GATA4 and 6 required
lower concentrations. GATA MOs inhibited Sox17
and HNF1ß induction
by activin at low and high concentrations in the order:
GATA6>GATA4>GATA5. Together with the timing of their expression and the
effects of GATA MOs in vivo, these observations identify GATA6 as the
predominant GATA factor in the maintenance of endodermal gene expression by
TGFß signaling in gastrulating embryos. In addition, examination of gene
expression and morphology in later embryos, revealed GATA5 and 6 as the most
critical for the development of the gut and the liver.
Key words: Activin, Animal caps, Cycloheximide, Emetine, Endoderm, GATA factors, Gut, HNF1ß, HNF3ß, Liver, Nodal-related, Sox17
, TGFß, Transcription, VegT, Xenopus
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