To directly monitor the glial activity in the CNS of the pond snail, Lymnaea stagnalis, we optically measured the electrical responses in the cerebral ganglion and median lip nerve to electrical stimulation of the distal end of the median lip nerve. Using a voltage-sensitive dye, RH155, we detected a composite depolarizing response in the cerebral ganglion, which consisted of a fast transient depolarizing response corresponding to a compound action potential and a slow depolarizing response. The slow depolarizing response was observed more clearly in an isolated median lip nerve and also detected by extracellular recording. In the median lip nerve preparation, the slow depolarizing response was suppressed by an L-type Ca2+ channel blocker, nifedipine, and was resistant to tetrodotoxin and Na+-free conditions. Together with the fact that a delay from the compound action potential to the slow depolarizing response was not constant, these results suggested that the slow depolarizing response was not a postsynaptic response. Because the signals of the action potentials appeared on the saturated slow depolarizing responses during repetitive stimulation, the slow depolarizing response was suggested to originate from glial cells. The contribution of the L-type Ca2+ current to the slow depolarizing response was confirmed by optical recording in the presence of Ba2+ and also supported by intracellular Ca2+ measurement.Our results suggested that electrical stimulation directly triggers glial Ca2+ entry through L-type Ca2+ channels, providing evidence for the generation of glial depolarization independent of neuronal activity in invertebrates. Copyright (C) 2000 IBRO.
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