Journal of Neurophysiology, Vol 70, Issue 2 710-722, Copyright © 1993 by APS

ARTICLES

A caffeine- and ryanodine-sensitive Ca2+ store in avian sensory neurons

A. Ivanenko, M. D. Baring, J. A. Airey, J. L. Sutko and J. L. Kenyon
Department of Physiology, University of Nevada School of Medicine, Reno 89557.

1. We identified and studied the function of ryanodine receptors in neurons isolated from dorsal root ganglia (DRG) of 10-day-old chick embryos. 2. A monoclonal antibody (mAb 34C) that recognizes all known ryanodine receptor isoforms in skeletal and cardiac muscle and CNS identified ryanodine receptor-like immunoreactivity in cultured DRG neurons. 3. Using the permeabilized patch technique to record membrane currents, we found that calcium currents were followed by a current with characteristics of a Ca(2+)-activated Cl- current (ICl(Ca)) in approximately two-thirds of the neurons. In these cells, acute application of 10 mM caffeine activated a similar ICl(Ca) and this effect was inhibited by 10 microM ryanodine. The activation of ICl(Ca) by caffeine was not dependent on extracellular Ca2+. These data suggest that caffeine raises intracellular free Ca2+ (Cai2+) by activating the release of Ca2+ from an intracellular store and that this Ca2+ activates the membrane conductance responsible for ICl(Ca). 4. The magnitude of ICl(Ca) activated by depolarization was not affected by ryanodine, implying that the Ca2+ that activates ICl(Ca) in this protocol is supplied by the Ca2+ current without amplification by a ryanodine-sensitive mechanism such as Ca(2+)-induced Ca2+ release. 5. We also used indo-1 to measure Cai2+ in DRG neurons. Ten millimolar caffeine caused a transient increase in Cai2+ that was inhibited by 10 microM ryanodine. 6. The ability of caffeine to elevate Cai2+ and activate ICl(Ca) was reduced at higher temperatures, suggesting increased Ca2+ sequestration. 7. These data demonstrate the existence of an intracellular store of Ca2+ that can be mobilized by a caffeine- and ryanodine-sensitive mechanism. The release of Ca2+ from this store can elevate Cai2+ and modulate membrane conductances.

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