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Regulatory volume decrease by SV40-transformed rabbit corneal epithelial cells requires ryanodine-sensitive

X Wu1, H Yang, P Iserovich

  • 1Department of Biological Sciences, State University of New York, College of Optometry, New York, NY 10010, USA.

The Journal of Membrane Biology
|July 15, 1997
PubMed
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Hypotonic stress triggers cell swelling and a calcium transient in tRCE cells, which is essential for regulatory volume decrease (RVD). Ryanodine-sensitive calcium release precedes increased calcium influx, both critical for RVD.

Area of Science:

  • Cell Biology
  • Physiology
  • Biochemistry

Background:

  • Cell volume regulation is crucial for cellular function.
  • Hypotonicity induces cell swelling, necessitating a regulatory volume decrease (RVD).
  • Intracellular calcium ([Ca2+]i) dynamics are implicated in cellular responses to osmotic stress.

Purpose of the Study:

  • To characterize the relationship between cell volume changes and intracellular calcium transients during hypotonic exposure.
  • To elucidate the temporal sequence of calcium influx and release mechanisms involved in RVD.
  • To identify the specific calcium stores and pathways critical for RVD in tRCE cells.

Main Methods:

  • Light scattering measurements to assess relative cell volume changes.
  • Fura2-fluorescence single-cell imaging to monitor intracellular calcium ([Ca2+]i) dynamics.

Related Experiment Videos

  • Mn2+ quenching assays to quantify plasmalemma calcium permeability and differentiate calcium influx/release.

    • Main Results:

    • Hypotonicity caused rapid cell swelling followed by 97% volume recovery (RVD) with a characteristic time of 5.9 min.
    • [Ca2+]i increased by 216% within 30 seconds, returning to baseline over 100 seconds.
    • RVD and [Ca2+]i transients were inhibited by blocking calcium influx (Ni2+), depleting intracellular stores (CPA), or inhibiting ryanodine-sensitive release.

    Conclusions:

    • Ryanodine-sensitive calcium release from intracellular stores precedes and triggers an increase in plasmalemma calcium influx.
    • Both intracellular calcium release and subsequent influx are essential components for initiating regulatory volume decrease (RVD).
    • This study provides the first temporal characterization of calcium signaling events driving RVD in response to hypotonic stress.