Translation and functional expression of cell-cell channel mRNA in Xenopus oocytes

Rudolf Werner, Todd Miller, Roobik Azarnia, Gerhard Dahl

Research output: Contribution to journalArticlepeer-review

31 Scopus citations

Abstract

mRNA from estrogen-stimulated rat myometrium, a tissue known to upregulate cell-cell channels in response to this hormone, was microinjected into Xenopus laevis oocytes. The oocytes had been freed from covering layers of follicle cells and vitelline to allow direct cell membrane interactions when paired. About 4 hours after the mRNA injection, paired oocytes become electrically coupled. This coupling was due to the presence of typical cell-cell channels characterized by size-limited intercellular tracer flux, the presence of gap junctions at the oocyte-oocyte interface, and the reversible uncoupling that occurred in the presence of carbon dioxide. The induction of new cell-cell channels in the oocyte membrane was observed against a zero background or a low level of endogenous coupling, depending on the maturation stage of the oocytes. The time course of development of cell-cell coupling after the microinjection of mRNA was determined. The mRNA capable of inducing cell-cell coupling was confined to an intermediate size class when fractionated on a sucrose gradient.

Original languageEnglish (US)
Pages (from-to)253-268
Number of pages16
JournalThe Journal of Membrane Biology
Volume87
Issue number3
DOIs
StatePublished - Oct 1 1985

Keywords

  • cell-cell channels
  • in vivo translation
  • mRNA
  • size fractionation
  • Xenopus oocytes

ASJC Scopus subject areas

  • Physiology
  • Cell Biology
  • Biophysics

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