Release and sequestration of calcium by ryanodine-sensitive stores in rat hippocampal neurones

J Physiol. 1997 Jul 1;502 ( Pt 1)(Pt 1):13-30. doi: 10.1111/j.1469-7793.1997.013bl.x.


1. The properties of ryanodine-sensitive Ca2+ stores in CA1 pyramidal cells were investigated in rat hippocampal slices by using whole-cell patch-clamp recordings combined with fura-2-based fluorometric digital imaging of cytoplasmic Ca2+ concentration ([Ca2+]i). 2. Brief pressure applications of caffeine onto the somata of pyramidal cells caused large transient increases in [Ca2+]i (Ca2+ transients) of 50-600 nM above baseline. 3. The Ca2+ transients evoked by caffeine at -60 mV were not associated with an inward current, persisted after blocking voltage-activated Ca2+ currents and were completely blocked by bath-applied ryanodine. Similar transients were also evoked at +60 mV. Thus, these transients reflect Ca2+ release from intracellular ryanodine-sensitive Ca2+ stores. 4. The Ca2+ transients evoked by closely spaced caffeine pulses rapidly decreased in amplitude, indicating progressive depletion of the Ca2+ stores. The amplitude of the Ca2+ transients recovered spontaneously with an exponential time constant of 59 s. Recovery was accelerated by depolarization-induced elevations in [Ca2+]i and blocked by cyclopiazonic acid (CPA) and thapsigargin, indicating that store refilling is mediated by endoplasmic reticulum Ca(2+)-ATPases. 5. Even without prior store depletion the caffeine-induced Ca2+ transients disappeared after 6 min exposure to CPA, suggesting that ryanodine-sensitive Ca2+ stores are maintained at rest by continuous Ca2+ sequestration. 6. Caffeine-depleted Ca2+ stores did not refill in Ca(2+)-free saline, suggesting that the refilling of the stores depends upon Ca2+ influx through a 'capacitative-like' transmembrane influx pathway operating at resting membrane potential. The refilling of the stores was also blocked by Ni2+ and gallopamil (D600). 7. Elevations of basal [Ca2+]i produced by bath-applied KCl markedly potentiated (up to 6-fold) the caffeine-induced Ca2+ transients. The degree of potentiation was positively related to the increase in basal [Ca2+]i. The Ca2+ transients remained potentiated up to 9 min after reversing the KCl-induced [Ca2+]i increase. Thus, the ryanodine-sensitive Ca2+ stores can 'overcharge' when challenged with an increase in [Ca2+]i and slowly discharge excess Ca2+ after basal [Ca2+]i returns to its resting level. 8. Pressure applications of caffeine onto pyramidal cell dendrites evoked local Ca2+ transients similar to those separately evoked in the respective somata. Thus, dendritic ryanodine-sensitive Ca2+ stores are also loaded at rest and can function as independent compartments. 9. In conclusion, the ryanodine-sensitive Ca2+ stores in hippocampal pyramidal neurones contain a releasable pool of Ca2+ that is maintained by a Ca2+ entry pathway active at subthreshold membrane potentials. Ca2+ entry through voltage-gated Ca2+ channels transiently overcharges the stores. Thus, by acting as powerful buffers at rest and as regulated sources during activity, Ca2+ stores may control the waveform of physiological Ca2+ signals in CA1 hippocampal pyramidal neurones.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Caffeine / pharmacology
  • Calcium / metabolism*
  • Calcium / pharmacology
  • Calcium Channels / metabolism*
  • Calmodulin-Binding Proteins / metabolism
  • Central Nervous System Stimulants / pharmacology
  • Dendrites / chemistry
  • Dendrites / metabolism
  • Enzyme Inhibitors / pharmacology
  • Hippocampus / cytology*
  • Hippocampus / physiology
  • Image Processing, Computer-Assisted
  • Indoles / pharmacology
  • Membrane Potentials / drug effects
  • Membrane Potentials / physiology
  • Muscle Proteins / metabolism*
  • Patch-Clamp Techniques
  • Potassium / metabolism
  • Pyramidal Cells / chemistry*
  • Pyramidal Cells / drug effects
  • Pyramidal Cells / ultrastructure
  • Rats
  • Rats, Wistar
  • Ryanodine Receptor Calcium Release Channel
  • Synapses / chemistry
  • Synapses / drug effects
  • Synapses / physiology
  • Thapsigargin / pharmacology


  • Calcium Channels
  • Calmodulin-Binding Proteins
  • Central Nervous System Stimulants
  • Enzyme Inhibitors
  • Indoles
  • Muscle Proteins
  • Ryanodine Receptor Calcium Release Channel
  • Caffeine
  • Thapsigargin
  • Potassium
  • Calcium
  • cyclopiazonic acid