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Related Experiment Videos

Calcium release-activated calcium current in rat mast cells

M Hoth1, R Penner

  • 1Department of Membrane Biophysics, Max-Planck-Institute for Biophysical Chemistry, Göttingen, Germany.

The Journal of Physiology
|June 1, 1993
PubMed
Summary

This study investigates the calcium release-activated calcium current (ICRAC) in rat peritoneal mast cells. Researchers found ICRAC is activated by store depletion and highly selective for calcium ions, with properties influenced by intracellular calcium buffering.

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Area of Science:

  • Cellular physiology
  • Ion channel biophysics
  • Calcium signaling

Background:

  • Intracellular calcium stores play a crucial role in regulating cellular processes.
  • Calcium release-activated calcium current (ICRAC) is a key mechanism for calcium influx following store depletion.
  • Understanding ICRAC biophysical properties is essential for elucidating mast cell function.

Purpose of the Study:

  • To characterize the biophysical properties of the calcium current activated by depletion of intracellular calcium stores in rat peritoneal mast cells.
  • To investigate the activation mechanisms, kinetics, and ion selectivity of this current.
  • To determine the influence of intracellular calcium buffering on ICRAC.

Main Methods:

  • Whole-cell patch clamp electrophysiology to record membrane currents.

Related Experiment Videos

  • Fura-2 fluorescence measurements to quantify intracellular calcium concentration ([Ca2+]i).
  • Induction of store depletion via InsP3 infusion, ionomycin application, or calcium chelators (EGTA, BAPTA).

    • Main Results:

    • ICRAC activation by store depletion showed a rapid onset (4-14 s) and monoexponential time course (20-30 s).
    • Current amplitude varied with intracellular buffers (EGTA vs. BAPTA), suggesting calcium-dependent inactivation.
    • ICRAC exhibited high selectivity for Ca2+ over other cations and was dependent on extracellular Ca2+ concentration (KD ≈ 3.3 mM).
    • Current amplitude was relatively insensitive to extracellular Mg2+ but dose-dependently inhibited by Cd2+.

    Conclusions:

    • The study elucidates key biophysical characteristics of ICRAC in rat mast cells.
    • ICRAC activation and properties are significantly influenced by intracellular calcium buffering dynamics.
    • Findings contribute to a deeper understanding of calcium signaling pathways in immune cells.