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Related Experiment Videos

Spontaneous calcium oscillations in embryonic stem cell-derived primitive endodermal cells

H Sauer1, C Hofmann, M Wartenberg

  • 1Institute for Neurophysiology, University of Cologne, Germany. hs@physiologie.uni-koeln.de

Experimental Cell Research
|February 11, 1998
PubMed
Summary

Spontaneous intracellular calcium (Ca2+) oscillations in mouse embryoid bodies increase in frequency and decrease in duration during differentiation. These oscillations are crucial for endo/exocytotic vesicle transport.

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Area of Science:

  • Developmental Biology
  • Cell Biology
  • Stem Cell Research

Background:

  • Embryoid bodies mimic early embryonic development, including endoderm differentiation.
  • Primitive endoderm differentiates into visceral and parietal endoderm.

Purpose of the Study:

  • To investigate spontaneous intracellular calcium ([Ca2+]i) oscillations during endoderm differentiation in mouse embryoid bodies.
  • To elucidate the signaling pathways and mechanisms regulating these Ca2+ oscillations.

Main Methods:

  • Confocal laser-scanning microscopy was used to monitor [Ca2+]i oscillations in primitive endodermal cells.
  • Pharmacological agents were employed to probe the dependence of oscillations on extracellular Ca2+, intracellular stores, and signaling pathways (PLC, PKC, InsP3).

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Main Results:

  • [Ca2+]i oscillation frequency increased, while duration decreased during differentiation from day 4 to day 19.
  • Oscillations required extracellular Ca2+ and intracellular Ca2+ release, modulated by phospholipase C (PLC) and protein kinase C (PKC) pathways.
  • Inhibition of endo/exocytotic pathways affected spiking activity, and absence of extracellular Ca2+ or presence of brefeldin A/thapsigargin abolished endocytotic vesicles.

Conclusions:

  • Spontaneous [Ca2+]i oscillations are a key feature of endoderm differentiation in embryoid bodies.
  • These oscillations are regulated by complex signaling pathways involving Ca2+ influx, release, PLC, and PKC.
  • Oscillating [Ca2+]i transients appear to play a role in the exo/endocytotic vesicle shuttle during differentiation.