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Related Experiment Videos

Smooth muscle gelsolin and a Ca2+-sensitive contractile cell model.

K Kanno1, Y Sasaki

  • 1Bio-Science Laboratory, Asahi Chemical Industry Co., Ltd, Nobeoka, Japan.

Journal of Cellular Physiology
|April 1, 1989
PubMed
Summary

Smooth muscle gelsolin severs actin filaments in human lung cells, inhibiting contraction. Prolonged high calcium levels are needed to activate gelsolin and disrupt stress fibers for cell relaxation.

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Area of Science:

  • Cell Biology
  • Biochemistry
  • Muscle Physiology

Background:

  • Smooth muscle gelsolin, a 90-kDa protein, was purified from bovine aorta.
  • Gelsolin's role in cell contraction and actin dynamics was investigated.
  • Human lung fibroblast MRC-5 cells were used as a model system.

Purpose of the Study:

  • To investigate the role of smooth muscle gelsolin in cell contraction.
  • To determine the effect of gelsolin on actin filament integrity in MRC-5 cells.
  • To elucidate the relationship between calcium levels, gelsolin activity, and cell contractility.

Main Methods:

  • Purification of smooth muscle gelsolin.
  • Immunofluorescence microscopy with phalloidin and anti-gelsolin antibody.
  • Immunoblotting and indirect immunofluorescence techniques.

Related Experiment Videos

  • Treatment of MRC-5 cells with purified gelsolin and calcium ionophore A23187.

    • Main Results:

    • Gelsolin was found along stress fibers in growth-arrested MRC-5 cells.
    • Purified gelsolin dose-dependently destroyed stress fibers and inhibited contractility.
    • Extracellular calcium-dependent contraction occurred with A23187, but without massive stress fiber destruction.
    • Endogenous gelsolin appeared inactive under transient calcium increases.

    Conclusions:

    • Continuous actin filaments in stress fibers are essential for cell contraction.
    • High cytoplasmic calcium concentrations maintained for minutes are required to activate endogenous gelsolin and disrupt stress fibers, inhibiting contraction.
    • Gelsolin-mediated stress fiber destruction is a key mechanism for cell relaxation under sustained calcium elevation.