Natural sounds, such as vocal communication sounds of many animal species typically occur as sequential sound pulses. Therefore, the response size of auditory neurons to a sound pulse would be inevitably affected when the sound pulse is preceded and succeeded by another sound pulse (i.e., forward and backward masking). The present study presents data to show that increasing strength of GABAergic inhibition relative to excitation contributes to decreasing response size and sharpening of duration selectivity of bat inferior collicular (IC) neurons to sound pulses in rapid sequences. The response size in number of impulses and duration selectivity of IC neurons were studied with a pulse train containing 9 sound pulses. A family of duration tuning curves was plotted for IC neurons using the number of impulses discharged to each presented sound pulse against pulse duration. Our data show that the response size of IC neurons progressively decreased and duration selectivity increased when determined with sequentially presented sound pulses. This variation in the response size and duration selectivity of IC neurons with sequentially presented sound pulses was abolished or reduced during bicuculline and GABA application. Bicuculline application increased the response size and broadened the duration tuning curve of IC neurons while GABA application produced opposite results. Possible mechanisms underlying increasing strength of GABAergic inhibition with sequentially presented sound pulses are presented. Biological significance of these findings in relation to acoustic signal processing is also discussed.