The acoustic startle response (ASR) depends on stimulus parameters such as duration, intensity and particularly on the stimulus rise time. The aim of our study was to determine to what extent the ASR parameters are affected by the spectral characteristics of the stimulus. Therefore, in this experiment the amplitude and the latency of the acoustic startle reflex were assessed for a fixed pulse duration and for a variety of stimulus frequencies ranging between 3 and 23 kHz. The ASRs were studied in 11 adult hooded rats exposed to 2 ms (120 dB SPL) tone pulses of different frequencies presented in random order, with or without 70 dB white noise background. Statistical analysis of the data revealed significant differences between ASR amplitudes for different frequencies. In our experimental situation the rats responded more readily to a low frequency stimulus. The startle amplitude decreased with tonal frequencies and distinguishable difference were seen for 3, 7, and 10 kHz pulses. However, such differences were not readily observed for higher frequencies i.e. 15, 20, 23 kHz. The same pattern of differences was observed when the acoustic stimulus was presented with the white noise background. The observed differences may be attributed, firstly, to a spectral characteristic of the stimulus and thus to an audibility in rats and secondly to a behavioral meaning of a stimulus of a different frequency. cy.