Spontaneous Ca2+ transients in developing hippocampal pyramidal cells.

Abstract

To determine the spatiotemporal pattern of hippocampal pyramidal cell activity during development, we examined cytosolic Ca2+ dynamics in tissue slices derived from early postnatal rats. After a brief (12-60 h) culture period, slices were stained with a calcium-sensitive dye, Fluo-3. Fluorescence imaging of the Fluo-3-stained slices with a scanning laser confocal microscope afforded simultaneous observation of many cells at high spatial resolution. Time-lapse imaging revealed spontaneous Ca2+ transients in the somata dendrites of many pyramidal cells in areas CA1 and CA3. For the most part, Ca2+ activity in neighboring pyramidal cells appeared to be uncorrelated, although we occasionally observed synchronous Ca2+ transients in adjacent cells. The transients were blocked by both tetrodotoxin (1 microM) and a mixture of the glutamate receptor antagonists, APV (50 microM) + CNQX (10 microM). Thus, spontaneous Ca2+ transients appear to be a consequence of activity-dependent release of glutamate acting postsynaptically through ionotropic glutamate receptors. Although gamma-aminobutyric acid (GABA) is thought to be an excitatory neurotransmitter during hippocampal development (Cherubini et al., 1991, Trends Neurosci. 14 (12):515-519), the spontaneous Ca2+ transients were not blocked by the GABAA receptor antagonist picrotoxin (100 microM). Furthermore, application of GABA (50 microM) abolished the spontaneous Ca2+ events, possibly via GABAB receptor-mediated inhibition of postsynaptic cells. The present results join other recent observations suggesting that isolated neural tissues support spontaneous activity, although the patterns and mechanisms of the activity reported here appear to differ from those of previous studies. Differences in the patterns of spontaneous activity during development may contribute to variations in the functional organization of different regions of CNS tissue.