1. Experiments were performed to identify the neurotransmitter released from hair cells of chick cochlea. An isolated hair cell was closely apposed to a cultured granule cell of the rat cerebellum, and both cells were whole-cell voltage clamped by utilizing a nystatin perforated patch technique. 2. Depolarization of hair cells to potentials more positive than -20 mV induced currents in the granule cell in a 10 mM Ca2+ extracellular medium. Amplitudes of induced currents were dependent on the membrane potential of granule cells and showed an outward-going rectification. The induced current in granule cells was reversibly suppressed by a local application of 2-amino-5-phosphonovalerate (APV), which indicates that the current was generated through the activation of an NMDA subtype of the glutamate receptor expressed on the granule cell. 3. The current amplitude of the granule cell was dependent on the size of hair cell depolarization. The size of current induced in a granule cell held at +55 mV was progressively increased with hair cell depolarization from -20 to +10 mV. At more positive potentials, the current amplitude was decreased. This voltage dependence was similar to but did not exactly match that of Ca2+ currents in the hair cell. The granule cell current appeared at more positive membrane potentials than the Ca2+ current in hair cells. 4. When intracellular Ca2+ concentration was increased by UV irradiation of the hair cell loaded with a caged Ca2+ compound, nitr-5, the closely apposed granule cell generated an outward current when voltage clamped at +55 mV. 5. These observations (paragraphs 2-4) imply that the most likely neurotransmitter released from the hair cell at its synapse with the afferent nerve terminal is glutamate.