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Volume 17, Number 1, Issue of January 1, 1997 pp. 160-170
Copyright ©1997 Society for Neuroscience

Low-Threshold Ca²⁺ Currents in Dendritic Recordings from Purkinje Cells in Rat Cerebellar Slice Cultures

Received July 12, 1996; revised Oct. 15, 1996; accepted Oct. 22, 1996.

Didier Mouginot, Jean-Louis Bossu, and Beat H. Gähwiler

Brain Research Institute, University of Zurich, CH-8029 Zurich, Switzerland

Voltage-dependent Ca²⁺ conductances were investigated in Purkinje cells in rat cerebellar slice cultures using the whole-cell and cell-attached configurations of the patch-clamp technique. In the presence of 0.5 mM Ca²⁺ in the extracellular solution, the inward current activated with a threshold of 55 ± 1.5 mV and reached a maximal amplitude of 2.3 ± 0.4 nA at 31 ± 2 mV. Decay kinetics revealed three distinct components: a fast (24.6 ± 2 msec time constant), a slow (304 ± 46 msec time constant), and a nondecaying component. Rundown of the slow and sustained components of the current, or application of antagonists for the P/Q-type Ca²⁺ channels, allowed isolation of the fast-inactivating Ca²⁺ current, which had a threshold for activation of 60 mV and reached a maximal amplitude of 0.7 nA at a membrane potential of 33 mV. Both activation and steady-state inactivation of this fast-inactivating Ca²⁺ current were described with Boltzmann equations, with half-activation and inactivation at 51 mV and 86 mV, respectively. This Ca²⁺ current was nifedipine-insensitive, but its amplitude was reduced reversibly by bath-application of NiCl₂ and amiloride, thus allowing its identification as a T-type Ca²⁺ current. Channels with a conductance of 7 pS giving rise to a fast T-type ensemble current (insensitive to -Aga-IVA) were localized with a high density on the dendritic membrane. Channel activity responsible for the ensemble current sensitive to -Aga-IVA was detected with 10 mM Ba²⁺ as the charge carrier. These channels were distributed with a high density on dendritic membranes and in rare cases were also seen in somatic membrane patches.

Key words: T-type calcium channels; cerebellum; Purkinje cells; patch clamp; dendritic recordings; cerebellar slice cultures

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