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Related Experiment Videos

Functionally separate intracellular Ca2+ stores in smooth muscle.

E R Flynn1, K N Bradley, T C Muir

  • 1Neuroscience and Biomedical Systems, Institute of Biomedical and Life Sciences, West Medical Bldg., University of Glasgow, Glasgow G12 8QQ, United Kingdom.

The Journal of Biological Chemistry
|July 31, 2001
PubMed
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Smooth muscle calcium signaling involves inositol 1,4,5-trisphosphate receptors (Ins(1,4,5)P(3)R) and ryanodine receptors (RyR) interacting with distinct sarcoplasmic reticulum (SR) stores. These findings reveal two functionally separate SR Ca(2+) stores with different refilling mechanisms.

Area of Science:

  • Cellular Biology
  • Physiology
  • Biochemistry

Background:

  • Cytosolic Ca(2+) concentrations ([Ca(2+)](c)) in smooth muscle are regulated by inositol 1,4,5-trisphosphate receptors (Ins(1,4,5)P(3)R) and ryanodine receptors (RyR) on the sarcoplasmic reticulum (SR).
  • The spatial organization and interplay of Ins(1,4,5)P(3)R and RyR on SR Ca(2+) stores critically influence Ca(2+) signaling dynamics.
  • The precise arrangement of these receptors on common or distinct SR stores remains unresolved.

Purpose of the Study:

  • To investigate the functional arrangement and Ca(2+) store relationships of Ins(1,4,5)P(3)R and RyR in isolated colonic myocytes.
  • To elucidate how the organization of these receptors impacts Ca(2+) release and signaling.

Main Methods:

  • Utilized isolated single voltage-clamped colonic myocytes.

Related Experiment Videos

  • Employing caffeine application to deplete ryanodine-sensitive stores.
  • Manipulating external Ca(2+) to assess Ins(1,4,5)P(3)-sensitive store dependence.
  • Inhibition of SR Ca(2+) ATPase with thapsigargin and cyclopiazonic acid to study store refilling.

    • Main Results:

    • Depletion of ryanodine-sensitive stores abolished Ins(1,4,5)P(3)-induced Ca(2+) release, suggesting a shared Ca(2+) store.
    • Ins(1,4,5)P(3)R-mediated Ca(2+) release did not trigger Ca(2+)-induced Ca(2+) release at RyR.
    • Depletion of Ins(1,4,5)P(3)-sensitive stores resulted in a minor reduction in caffeine-evoked transients, indicating not all RyR reside on the common store.
    • Ins(1,4,5)P(3)-sensitive stores depend on external Ca(2+) for refilling, unlike a subset of RyR-containing stores refilled from the cytosol.

    Conclusions:

    • Evidence supports the existence of two functionally distinct Ca(2+) stores within the SR.
    • One store appears to contain only RyR and is refilled from the cytosol ([Ca(2+)](c)).
    • A second, distinct store contains both Ins(1,4,5)P(3)R and RyR and relies on external Ca(2+) for replenishment.