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The Journal of Neuroscience, August 4, 2004, 24(31):6946-6957; doi:10.1523/JNEUROSCI.1397-04.2004

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Cellular/Molecular
Action Potential-Evoked and Ryanodine-Sensitive Spontaneous Ca²⁺ Transients at the Presynaptic Terminal of a Developing CNS Inhibitory Synapse

Rossella Conti,¹ Yusuf P. Tan,² and Isabel Llano¹

¹Laboratoire de Physiologie Cérébrale, Université Paris 5, 75006 Paris, France, and ²Bogazici Universitesi, Bebek, Istanbul 80815, Turkey

The existence of spontaneous calcium transients (SCaTs) dependent on intracellular store activation has been reported in putative axonal terminals of cerebellar basket interneurons. We used the two-photon imaging technique to optically identify basket terminals in acute cerebellar slices of young rats (11-16 d old) and study the properties of SCaTs unambiguously localized in these regions. The whole-cell recording configuration and preloading technique were alternatively used to load the calcium-dependent dye in the interneuron and compare SCaTs with action potential evoked calcium transients. SCaTs were observed in the basket terminals at frequencies that were significantly increased after bath application of 10 µM ryanodine and did not depend on P/Q- or N-type voltage-dependent calcium channel activation. They originated at specific sites where bursts of events with temporal separation as small as 200 msec could be generated. Their sites of origin were spaced on average 6 µm apart and were preferentially located near axonal endings. SCaTs had amplitudes comparable with those of Ca²⁺ rises evoked by single action potentials that lead to release of neurotransmitter, as confirmed by parallel recordings of preloaded terminals and evoked IPSCs in the postsynaptic Purkinje cells. These results support the hypothesis that SCaTs at basket terminals underlie the large miniature IPSCs characteristic of Purkinje cells.

Key words: confocal imaging; presynaptic terminal; ryanodine; calcium stores; cerebellum; GABAergic

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Received Dec 20, 2003; revised June 18, 2004; accepted June 18, 2004.

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