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Related Experiment Videos

Swelling-induced changes in cytosolic [Ca2++] in insulin-secreting cells: a role in regulatory volume decrease?

E A Sheader1, P D Brown, L Best

  • 1Department of Medicine, University of Manchester, M13 9WL, Manchester, UK.

Molecular and Cellular Endocrinology
|July 31, 2001
PubMed
Summary
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Hypotonic cell swelling in insulin-secreting cells activates chloride and potassium channels for regulatory volume decrease (RVD). While calcium influx occurs, it is not essential for RVD, which relies on low resting intracellular calcium.

Area of Science:

  • Cell physiology
  • Ion channel function
  • Endocrinology

Background:

  • Insulin-secreting cells swell in hypotonic solutions, initiating regulatory volume decrease (RVD).
  • Previous work identified a chloride conductance activated during RVD.
  • The role of intracellular calcium (Ca(2+)) in this process remained unclear.

Purpose of the Study:

  • Investigate the involvement of cytosolic Ca(2+) changes in hypotonic swelling and RVD of insulinoma cells.
  • Determine the specific ion channels and Ca(2+) signaling pathways mediating RVD.

Main Methods:

  • Utilized RINm5F insulinoma cells.
  • Applied hypotonic solutions to induce cell swelling.
  • Measured cytosolic Ca(2+) using fluorescence indicators.

Related Experiment Videos

  • Assessed RVD and ion channel activity using pharmacological blockers (D600, Gd(3+), DIDS, TEA, IbTx, tolbutamide) and ion-free solutions.

    • Main Results:

    • Hypotonic swelling increased cytosolic Ca(2+), dependent on extracellular Ca(2+) and voltage-gated Ca(2+) channels, but not maxi-K(+) (BK(Ca)) channels.
    • RVD was impaired without extracellular Ca(2+), indicating Ca(2+) dependence.
    • RVD was inhibited by BK(Ca) channel blockers (TEA, IbTx) but not tolbutamide.
    • Cell swelling activated a K(+) conductance sensitive to TEA and IbTx.

    Conclusions:

    • Cell swelling activates volume-sensitive anion channels, leading to depolarization and Ca(2+) influx.
    • RVD is Ca(2+)-dependent, requiring low resting intracellular Ca(2+), but the swelling-induced Ca(2+) increase is not essential.
    • RVD involves simultaneous activation of Cl(-) and K(+) channels, with BK(Ca) channels being the primary K(+) conductance.