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General Transcription Factors

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Tissue-specific transcription factors contribute to diverse cellular functions in mammals. For example, the gene for beta globin, a major component of hemoglobin, is present in all cells of the body. However, it is only expressed in red blood cells because the transcription factors that can bind to the promoter sequences of the beta globin gene are only expressed in these cells. Tissue-specific transcription factors also ensure that mutations in these factors may impair only the function of...
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Stem cell-like Xenopus Embryonic Explants to Study Early Neural Developmental Features In Vitro and In Vivo
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Functional differences between Tcf1 isoforms in early Xenopus development.

Giulietta Roël1, Olaf Van Den Broek, Olivier Destrée

  • 1Hubrecht Institute, Utrecht, the Netherlands.

The International Journal of Developmental Biology
|March 14, 2017
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Summary

Different Tcf1 isoforms in Xenopus embryos cause distinct developmental defects. The C-clamp is crucial for proper axis formation and gene expression, with its absence leading to severe developmental issues.

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Area of Science:

  • Developmental Biology
  • Molecular Biology
  • Genetics

Background:

  • Four Tcf1 isoforms (B, C, D, E) exist in Xenopus gastrula embryos, showing high conservation across species.
  • Functional differences between these Tcf1 isoforms during early Xenopus development were investigated.

Purpose of the Study:

  • To investigate the functional roles of different Tcf1 isoforms during early Xenopus development.
  • To determine the importance of the Tcf1 C-clamp domain in embryonic development and gene regulation.

Main Methods:

  • Overexpression of individual Tcf1 isoforms in Xenopus embryos.
  • Site-directed mutagenesis of the Tcf1 C-clamp domain.
  • Morpholino-based knockdown of the C-clamp exon.
  • In situ hybridization to analyze gene expression patterns (e.g., Lef1).

Main Results:

  • Overexpression of Tcf1 isoforms lacking a C-clamp (B, D) enhanced Wnt signaling, induced ectopic dorsal mesoderm, and caused antero-dorzalization.
  • Overexpression of the E-isoform (complete C-clamp) led to caudal truncation, not dorsal mesoderm induction.
  • Mutation of the C-clamp resulted in gain-of-function for ectopic organizer formation.
  • Depletion of the C-clamp exon severely shortened the AP-axis and impaired CNS, mesoderm, and blood vessel development.
  • Lef1 expression was downregulated in specific embryonic regions upon C-clamp depletion.

Conclusions:

  • The Tcf1 C-clamp is essential for proper anterior-posterior axis formation in Xenopus embryos.
  • The E-tail of Tcf1 is required for Lef1 expression and blood vessel formation in post-gastrula embryos.