Intracellular Ca(2+) release from endoplasmic reticulum regulates slow wave currents and pacemaker activity of interstitial cells of Cajal

Am J Physiol Cell Physiol. 2015 Apr 15;308(8):C608-20. doi: 10.1152/ajpcell.00360.2014. Epub 2015 Jan 28.


Interstitial cells of Cajal (ICC) provide pacemaker activity in gastrointestinal muscles that underlies segmental and peristaltic contractions. ICC generate electrical slow waves that are due to large-amplitude inward currents resulting from anoctamin 1 (ANO1) channels, which are Ca(2+)-activated Cl(-) channels. We investigated the hypothesis that the Ca(2+) responsible for the stochastic activation of ANO1 channels during spontaneous transient inward currents (STICs) and synchronized activation of ANO1 channels during slow wave currents comes from intracellular Ca(2+) stores. ICC, obtained from the small intestine of Kit(+/copGFP) mice, were studied under voltage and current clamp to determine the effects of blocking Ca(2+) uptake into stores and release of Ca(2+) via inositol 1,4,5-trisphosphate (IP3)-dependent and ryanodine-sensitive channels. Cyclocpiazonic acid, thapsigargin, 2-APB, and xestospongin C inhibited STICs and slow wave currents. Ryanodine and tetracaine also inhibited STICs and slow wave currents. Store-active compounds had no direct effects on ANO1 channels expressed in human embryonic kidney-293 cells. Under current clamp, store-active drugs caused significant depolarization of ICC and reduced spontaneous transient depolarizations (STDs). After block of ryanodine receptors with ryanodine and tetracaine, repolarization did not restore STDs. ANO1 expressed in ICC has limited access to cytoplasmic Ca(2+) concentration, suggesting that pacemaker activity depends on Ca(2+) dynamics in restricted microdomains. Our data from studies of isolated ICC differ somewhat from studies on intact muscles and suggest that release of Ca(2+) from both IP3 and ryanodine receptors is important in generating pacemaker activity in ICC.

Keywords: ANO1 channel; IP3 receptor; SERCA pump; ryanodine receptor.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • Anoctamin-1
  • Calcium / metabolism*
  • Calcium Channel Blockers / pharmacology
  • Calcium-Transporting ATPases / antagonists & inhibitors
  • Cells, Cultured
  • Chloride Channels / biosynthesis
  • Chloride Channels / metabolism*
  • Endoplasmic Reticulum / metabolism*
  • Enzyme Activation
  • Enzyme Inhibitors / pharmacology
  • HEK293 Cells
  • Humans
  • Indoles / pharmacology
  • Inositol 1,4,5-Trisphosphate / chemistry
  • Interstitial Cells of Cajal / metabolism*
  • Intestine, Small / cytology
  • Macrocyclic Compounds / pharmacology
  • Membrane Potentials / drug effects
  • Mice
  • Muscle Contraction / physiology
  • Myocytes, Smooth Muscle / metabolism
  • Oxazoles / pharmacology
  • Patch-Clamp Techniques
  • Ryanodine / pharmacology
  • Ryanodine Receptor Calcium Release Channel / metabolism*
  • Thapsigargin / pharmacology


  • ANO1 protein, mouse
  • Anoctamin-1
  • Calcium Channel Blockers
  • Chloride Channels
  • Enzyme Inhibitors
  • Indoles
  • Macrocyclic Compounds
  • Oxazoles
  • Ryanodine Receptor Calcium Release Channel
  • xestospongin C
  • Ryanodine
  • Thapsigargin
  • Inositol 1,4,5-Trisphosphate
  • Calcium-Transporting ATPases
  • Calcium
  • cyclopiazonic acid