The number of presynaptic vesicles that are immediately available for release, the readily releasable pool (RRP), is a strong determinant of synaptic strength and plasticity. The properties of the RRP in individual GABAergic synapses were examined in superior colliculus cultures. The RRP was depleted by high frequency trains and cumulative evoked IPSC amplitudes (CA) were calculated. The amplitude of monoquantal responses (q) was determined on the basis of mlPSC histograms. On average, the RRP, defined as CA/q, comprised about 10 vesicles. About 60% of the RRP could be released by a single stimulus. After depletion, the RRP was replenished with a time constant of about 14 s. These data provide information for further studies on the capacity of individual inhibitory synapses to modulate sensory information transfer.