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Journal of Neuroscience, Vol 15, 2592-2608, Copyright © 1995 by Society for Neuroscience
Inositol 1,4,5-trisphosphate and ryanodine receptor distributions and patterns of acetylcholine- and caffeine-induced calcium release in cultured mouse hippocampal neurons
KJ Seymour-Laurent and ME Barish
Division of Neurosciences, Beckman Research Institute of the City of Hope, Duarte, California 91010, USA.
The distributions of inositol 1,4,5-trisphosphate and ryanodine receptors
(InsP3R and RyR) and the patterns of increase in intracellular calcium ion
concentration ([Ca2+]i) elicited by their activation were compared in
cultured hippocampal neurons. InsP3R and RyR were labeled using specific
antibodies and formed small aggregations in the somata and dendrites of
pyramidally shaped neurons. Both receptors were densest in somata. In
dendrites the InsP3R and RyR were not distributed homogeneously; InsP3R was
found in all regions, while RyR was least dense in fine processes.
Increases in [Ca2+]i elicited by acetylcholine (to activate InsP3 receptors
via muscarinic receptors) and caffeine (to stimulate ryanodine receptors)
were measured in dendrites using Ca(2+)-sensitive fluorescent dyes and
confocal microscopy. Ca2+ responses to acetylcholine were transient and
observed in proximal and distal dendritic regions. In contrast,
caffeine-induced responses were sustained and restricted to proximal
dendrites. Thus the patterns of calcium release in fine dendrites mirrored
the distributions of InsP3R and RyR. Calcium responses to both
acetylcholine and caffeine were observed in the absence of external calcium
and thus were dependent on Ca2+ release. Ca2+ responses showed localized
fluctuations and variations in response delay times. Sequential activation
of InsP3R and RyR in somata resulted in mutual occlusion of Ca2+ release.
The existence of InsP3-gated and Ca(2+)- induced Ca2+ release as spatially
distinct, but mutually interacting, mechanisms may be important in the
generation of oscillations and propagating Ca2+ waves in somata and
dendrites of hippocampal neurons.
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