The physiological properties of Purkinje neurons play a central role in their ability to regulate information transfer through the cerebellum. A number of ion channels contribute to Purkinje neuron physiology including an abundance of P-type Ca2+ channels, particularly in the dendritic region. Purkinje neurons also express L-type Ca2+ channels both during development and in the mature state. However, a role for L-type channels in Purkinje neuron physiology has yet to be fully defined. In the current study we used physiological recordings from cultured Purkinje neurons and the L-type Ca2+ channel agonist S-(-)-Bay K to assess a potential role for L-type Ca2+ channels in spike firing. Results show that Bay K alters current-evoked spike firing in young, immature Purkinje neurons without dendritic structure and in older, more mature Purkinje neurons with dendritic structure. Bay K also enhanced Ca2+ signals associated with the current-evoked spike firing. The effect of Bay K was more prominent in the young Purkinje neurons than in the older Purkinje neurons, suggesting that L-type Ca2+ channels may be more important in the Purkinje neuron physiology during the early stages of development rather than at mature stages. In the older Purkinje neurons, immunohistochemical studies using antibodies to L-type Ca2+ channels showed more intense immunolabeling in the somatic region than in the dendritic region. This result suggests that L-type Ca2+ channels may play a more important role in somatic physiology than dendritic physiology, whereas P-type channels may play a more important role in dendritic physiology.