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The Journal of Neuroscience, October 1, 2000, 20(19):7394-7403
L-Type Calcium Channels Mediate Calcium Oscillations in Early Postnatal Purkinje Neurons
Patricia Liljelund, Jeffrey G. Netzeband, and Donna L. Gruol Department of Neuropharmacology, The Scripps Research Institute, La Jolla, California 92037
Ca2+ signaling is important in many fundamental neuronal processes including neurotransmission, synaptic plasticity, neuronal development, and gene expression. In cerebellar Purkinje neurons, Ca2+ signaling has been studied primarily in the dendritic region where increases in local Ca2+ have been shown to occur with both synaptic events and spontaneous electrical activity involving P-type voltage-gated Ca2+ channels (VGCCs), the predominant VGCC expressed by Purkinje neurons. Here we show that Ca2+ signaling is also a prominent feature of immature Purkinje neurons at developmental stages that precede expression of dendritic structure and involves L-type rather than P-type VGCCs. Immature Purkinje neurons acutely dissociated from postnatal day 4-7 rat pups exhibit spontaneous cytoplasmic Ca2+ oscillations. The Ca2+ oscillations require entry of extracellular Ca2+, are blocked by tetrodotoxin, are communicated to the nucleus, and correlate closely with patterns of endogenously generated spontaneous and evoked electrical activity recorded in the neurons. Immunocytochemistry showed that L-, N-, and P/Q-types of VGCCs are present on the somata of the Purkinje neurons at this age. However, only the L-type VGCC antagonist nimodipine effectively antagonized the Ca2+ oscillations; inhibitors of P/Q and N-type VGCCs were relatively ineffective. Release of Ca2+ from intracellular Ca2+ stores significantly amplified the Ca2+ signals of external origin. These results show that a somatic signaling pathway that generates intracellular Ca2+ oscillations and involves L-type VGCCs and intracellular Ca2+ stores plays a prominent role in the Ca2+ dynamics of early developing Purkinje neurons and may play an important role in communicating developmental cues to the nucleus.
Key words: cerebellum; development; acutely isolated neurons; Ca2+ signaling; nuclear Ca2+; intracellular Ca2+ stores
Copyright © 2000 Society for Neuroscience 0270-6474/00/20197394-10$05.00/0
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