Analysis of localization and reorganization of germ plasm in Xenopus transgenic line with fluorescence-labeled mitochondria

Dev Growth Differ. 2012 Oct;54(8):767-76. doi: 10.1111/dgd.12005.

Abstract

Germ plasm is found in germ-line cells of Xenopus and thought to include the determinant of primordial germ cells (PGCs). As mitochondria is abundant in germ plasm, vital staining of mitochondria was used to analyze the movement and function of germ plasm; however, its application was limited in early cleavage embryos. We made transgenic Xenopus, harboring enhanced green fluorescent protein (EGFP) fused to the mitochondria transport signal (Dria-line). Germ plasm with EGFP-labeled mitochondria was clearly distinguishable from the other cytoplasm, and retained mostly during one generation of germ-line cells in Dria-line females. Using the Dria-line, we show that germ plasm is reorganized from near the cell membrane to the perinuclear space at St. 9, dependent on the microtubule system.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Animals, Genetically Modified
  • Blastomeres / metabolism
  • Blastomeres / ultrastructure
  • Cell Nucleus / metabolism
  • Cells, Cultured
  • Cleavage Stage, Ovum / metabolism
  • Cytoplasm / ultrastructure*
  • Embryo, Nonmammalian / metabolism
  • Embryo, Nonmammalian / ultrastructure*
  • Female
  • Fluorescent Dyes
  • Gastrulation / physiology
  • Germ Cells / metabolism
  • Germ Cells / ultrastructure*
  • Green Fluorescent Proteins / genetics
  • Green Fluorescent Proteins / metabolism
  • Microtubules / metabolism*
  • Mitochondria / metabolism
  • Mitochondria / ultrastructure
  • Oocytes / cytology
  • Oocytes / metabolism
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / metabolism
  • Tubulin / metabolism
  • Xenopus laevis / embryology

Substances

  • Fluorescent Dyes
  • RNA, Messenger
  • Recombinant Fusion Proteins
  • Tubulin
  • enhanced green fluorescent protein
  • Green Fluorescent Proteins