Visualization of the Xenopus primordial germ cells using a green fluorescent protein controlled by cis elements of the 3' untranslated region of the DEADSouth gene

Mech Dev. 2006 Oct;123(10):746-60. doi: 10.1016/j.mod.2006.07.006. Epub 2006 Jul 21.

Abstract

We succeeded in visualization of the primordial germ cells (PGCs) in a living Xenopus embryo. The mRNA of the reporter Venus protein, fused to the 3' untranslated region (UTR) of DEADSouth, which is a component of the germ plasm in Xenopus eggs, was microinjected into the vegetal pole of fertilized eggs and then the cells with Venus fluorescence were monitored during development. The behavior of the cells was identical to that previously described for PGCs. Almost all Venus-expressing cells were Xdazl-positive in the stage 48 tadpoles, indicating that they were PGCs. In addition, we found three sub-regions (A, B and C) in the 3' UTR, which were involved in the PGC-specific expression of the reporter protein. Sub-region A, which was identified previously as a localization signal for the germ plasm during oogenesis, participated in anchoring of the mRNA at the germ plasm and the degradation of the mRNA in the somatic cells. Sub-regions B and C were also involved in anchoring of the mRNA at the germ plasm. Sub-region B participated in the enhancement of translation.

MeSH terms

  • 3' Untranslated Regions*
  • Amino Acid Sequence
  • Animals
  • Bacterial Proteins / genetics
  • Bacterial Proteins / metabolism*
  • Base Sequence
  • DNA Helicases / genetics*
  • DNA Helicases / metabolism
  • Female
  • Fluorescent Dyes / metabolism*
  • Genes, Reporter
  • Germ Cells / cytology*
  • In Situ Hybridization
  • Luminescent Proteins / genetics
  • Luminescent Proteins / metabolism*
  • Male
  • Microinjections
  • Molecular Sequence Data
  • Oocytes / metabolism*
  • RNA Stability
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / metabolism
  • Xenopus Proteins / genetics
  • Xenopus Proteins / metabolism
  • Xenopus laevis* / anatomy & histology
  • Xenopus laevis* / embryology
  • Xenopus laevis* / genetics

Substances

  • 3' Untranslated Regions
  • Bacterial Proteins
  • Fluorescent Dyes
  • Luminescent Proteins
  • Recombinant Fusion Proteins
  • Xenopus Proteins
  • yellow fluorescent protein, Bacteria
  • DNA Helicases