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Regulatory volume decrease and intracellular Ca2+ in murine neuroblastoma cells studied with fluorescent probes

J Altamirano1, M S Brodwick, F J Alvarez-Leefmans

  • 1Departamento de Neurobiología, Instituto Mexicano de Psiquiatría, México 14370, D.F. México.

The Journal of General Physiology
|August 5, 1998
PubMed
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Calcium ions (Ca2+) do not appear essential for triggering regulatory volume decrease (RVD) in neural cells, even though RVD is often accompanied by a transient Ca2+ increase. This finding challenges the established role of Ca2+ in cell volume regulation.

Area of Science:

  • Cell Biology
  • Physiology
  • Biochemistry

Background:

  • Regulatory Volume Decrease (RVD) is a critical cellular process for maintaining homeostasis in response to osmotic stress.
  • The role of intracellular calcium (Ca2+) as a second messenger in mediating RVD has been previously suggested but not definitively proven.
  • Neural cell lines offer a model system to investigate complex cellular signaling pathways.

Purpose of the Study:

  • To investigate the necessity of Ca2+ in triggering and regulating RVD in murine neural cell lines.
  • To simultaneously measure intracellular Ca2+ levels and cell volume changes during RVD.
  • To elucidate the precise role of Ca2+ in the inactivation of RVD.

Main Methods:

  • Utilized novel microspectrofluorimetric techniques for simultaneous single-cell measurement of intracellular Ca2+ ([Ca2+]i) and cell volume.

Related Experiment Videos

  • Employed fura-2 for ratiometric [Ca2+]i measurement and calcein as an independent volume probe.
  • Applied hyposmotic challenges and Ca2+ chelation (EGTA, BAPTA) to assess RVD responses.

    • Main Results:

    • Hyposmotic stress induced RVD, accompanied by a transient increase in [Ca2+]i that preceded RVD.
    • [Ca2+]i increase correlated with the degree of hyposmotic challenge and cell swelling.
    • Ca2+ chelation attenuated but did not abolish RVD, indicating a Ca2+-independent component.
    • Ca2+-independent RVD occurred even with decreased [Ca2+]i, and Ca2+ restoration did not affect RVD rate or extent.
    • RVD and [Ca2+]i increases were attenuated upon repeated hyposmotic challenges, irrespective of Ca2+ presence.

    Conclusions:

    • Simultaneous measurements demonstrate that an increase in intracellular Ca2+ is not essential for initiating RVD or its inactivation in these neuroblastoma cells.
    • The observed attenuation of RVD upon Ca2+ chelation may be due to secondary effects or a requirement for optimal, rather than essential, RVD.
    • These findings necessitate a re-evaluation of the role of Ca2+ as a primary mediator of RVD in neural cells.