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Related Experiment Video

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High Resolution Whole Mount In Situ Hybridization within Zebrafish Embryos to Study Gene Expression and Function
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Zebrafish Rnf111 is encoded by multiple transcripts and is required for epiboly progression and prechordal plate

Dmitri A Bessarab1, Sinnakaruppan Mathavan2, C Michael Jones1

  • 1Institute of Medical Biology, Agency for Science, Technology and Research (A⁎STAR), 8A Biomedical Grove, #06-06 Immunos, Singapore 138648, Singapore.

Differentiation; Research in Biological Diversity
|January 27, 2015
PubMed
Summary

Zebrafish Arkadia (RING finger 111) is crucial for early development, regulating Nodal signaling. Specific transcript variants are essential for epiboly progression during gastrulation.

Keywords:
Alternative splicingEpibolyEvolutionary conservationPrechordal plateZebrafishrnf111

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Area of Science:

  • Developmental Biology
  • Molecular Biology
  • Genetics

Background:

  • Arkadia (RING finger 111) is a nuclear E3 ubiquitin ligase involved in TGFβ/Nodal signaling.
  • Mouse Arkadia loss causes embryonic lethality due to impaired Nodal signaling.
  • The role of Arkadia in zebrafish development and its transcript variants is not well understood.

Purpose of the Study:

  • To investigate the function of zebrafish arkadia/rnf111 during early embryonic development.
  • To identify the roles of different arkadia/rnf111 transcript variants in developmental processes.
  • To understand Arkadia's contribution to Nodal signaling regulation in zebrafish.

Main Methods:

  • Isolation and characterization of zebrafish arkadia/rnf111 and its transcript variants.
  • Analysis of arkadia/rnf111 expression patterns during zebrafish embryogenesis.
  • Morpholino-mediated knockdown to assess arkadia/rnf111 function in blastula organization and epiboly.
  • Use of splice junction morpholinos to investigate the role of specific transcript variants.

Main Results:

  • Zebrafish arkadia/rnf111 has 5 transcript variants and is broadly expressed in early embryos, particularly in anterior mesendoderm.
  • Knockdown of arkadia/rnf111 disrupts early blastula organization and epiboly progression.
  • Specific arkadia/rnf111 transcript variants containing a 3' alternative exon are critical for late gastrula epiboly.

Conclusions:

  • Zebrafish Arkadia/Rnf111 plays a vital role in early embryonic organization and gastrulation movements.
  • Alternative splicing of arkadia/rnf111 generates functionally distinct protein isoforms.
  • These isoforms provide regulatory flexibility to the Nodal signaling pathway, essential for proper zebrafish development.