Abstract
The relationship between calcium conductances and developmental changes in the active and passive membrane properties of cerebellar Purkinje neurons from rats was studied in a culture model system by using current-clamp and voltage-clamp techniques. These cultures, at 6-21 d of age, represented the main period of morphological and physiological development of the Purkinje neuron. In the current-clamp studies, input resistance decreased and the current-voltage curve became more S-shaped as the neurons matured in culture. Spike-generating properties also changed. Immature Purkinje neurons without dendritic structure produced repetitive, fast TTX-sensitive simple spikes when stimulated electrically. The simple spike frequency increased with maturation. In older neurons (greater than or equal to 12 d in vitro) with well-developed dendritic structure, a burst event, the complex spike, preceded the repetitive simple spike firing. Magnesium (10 mM) and cadmium (50-100 microM), calcium channel blockers, antagonized the repetitive simple spike firing in both young and old neurons. The complex spike of the older neurons was also antagonized by magnesium (10 mM) but was resistant to cadmium (50-100 microM), suggesting that a pharmacologically distinct calcium conductance mediated this spike event. Whole-cell voltage-clamp recordings showed that the older Purkinje neurons expressed two calcium currents, a low-threshold rapidly inactivating calcium current resistant to cadmium (50-100 microM) and a high-threshold slowly inactivating calcium current antagonized by cadmium (50-100 microM). In young Purkinje neurons without dendritic structure (6-9 d in vitro), only the high-threshold calcium current was evident. The amplitude of this current increased approximately 50% during development. These results indicate that the developmental expression of calcium conductances plays a prominent role in the physiological maturation of the cultured Purkinje neurons, which closely simulate the physiologic cells they model. The high-threshold calcium conductance is expressed early in development and contributes to repetitive simple spike firing of both the young and old neurons. The low-threshold calcium conductance appears later in development, coincident with dendritic expression, and plays a major role in the generation of the complex spike.