Intracellular free Ca2+ concentration ([Ca2+]i) in spiral ganglion cells (SGC) of guinea pig was measured using the fura-2 fluorescence method. Application of high-K+ external solution increased [Ca2+]i. [Ca2+]i elevation was suppressed by either depletion of external Ca2+ or application of the dihydropyridine Ca2+ channel antagonist, nicardipine. One of the putative hair cell afferent nerve neurotransmitters, L-glutamate (Glu), induced [Ca2+]i elevation in SGC. Pharmacological studies suggested that the Glu receptor may be a non-NMDA subtype. Glu-induced response was suppressed by either depletion of external Ca2+ or application of nicardipine, indicating that Glu-induced [Ca2+]i elevation is mainly carried out by the voltage-dependent Ca2+ channel in SGC.