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Related Experiment Videos

Analysis of cell surface galactosyltransferase activity during mouse trophectodermal differentiation.

H J Hathaway1, L C Romagnano, B S Babiarz

  • 1Department of Biological Sciences, Rutgers University, Piscataway, New Jersey 08855-1059.

Developmental Biology
|August 1, 1989
PubMed
Summary

Mouse embryo trophoblast cells use a galactosyltransferase (GalTase)-mediated adhesion system involving lactosaminoglycans (LAGs). This system is crucial for cell-cell adhesion in the ectoplacental cone but is lost during differentiation into invasive giant cells.

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Area of Science:

  • Developmental Biology
  • Cell Biology
  • Glycobiology

Background:

  • The ectoplacental cone (EPC) of mouse embryos contains adhesive trophoblast cells that differentiate into invasive giant cells during implantation.
  • Adhesive trophoblast cells express cell surface lactosaminoglycans (LAGs), which are lost upon differentiation.
  • LAGs can act as substrates for cell surface galactosyltransferase (GalTase), mediating cell-cell adhesion.

Purpose of the Study:

  • To investigate the hypothesis that LAGs in the EPC serve as substrates for a GalTase-mediated cell-cell adhesion system.
  • To characterize the cell surface GalTase activity and its substrate in EPC trophoblast cells.
  • To determine the role of this adhesion system in trophoblast differentiation.

Main Methods:

  • Demonstration of cell surface GalTase activity in EPC trophoblast using radiolabeled galactose incorporation.

Related Experiment Videos

  • Immunofluorescence localization of GalTase to cell-cell contact areas in cultured EPC trophoblast.
  • Characterization of the GalTase substrate using size exclusion chromatography and enzymatic digestion.
  • Perturbation of the GalTase-substrate complex using specific inhibitors and antibodies to assess effects on cell adhesion.

    • Main Results:

    • Cell surface GalTase activity was detected on Day 7.5 EPC trophoblast.
    • Immunofluorescence localized GalTase to areas of cell-cell contact, with reduced labeling in differentiated giant cells.
    • A 15,000 Da glycopeptide, partially sensitive to endo-beta-galactosidase, was identified as the GalTase substrate containing LAG structures.
    • Perturbation of the GalTase-substrate complex disrupted cell-cell contacts in EPC outgrowths, and differentiated giant cells showed reduced sensitivity.

    Conclusions:

    • Adhesive EPC trophoblast cells utilize a GalTase-mediated cell-cell adhesion system involving LAGs.
    • This adhesion system is downregulated during the differentiation of trophoblast cells into invasive giant cells.
    • The findings elucidate a novel mechanism of cell adhesion critical for early embryonic development and implantation.