The acoustic startle response (ASR) can be enhanced by administration of footshocks (sensitization). The neural mechanisms underlying this effect are largely unknown. A previous electrophysiological study (Kungel et al., Brain Res., 643 (1994) 29-39) has shown that the neuropeptide substance P (SP) increases the responsiveness to acoustic stimuli of neurons in the caudal pontine reticular nucleus (PnC). Since the PnC is an important part of the primary acoustic startle circuit, we hypothesized that SP is involved in the enhancement of the ASR by electric footshocks. We tested this hypothesis in different experiments by locally injecting SP and SP-antagonists into the PnC of freely moving rats. The present data show that SP (0.5 pmol-1 nmol) locally injected into the PnC dose-dependently increases the amplitude of the ASR in rats. This effect was antagonized by pretreatment with the SP-antagonist CP-96,345. Furthermore, we show that the sensitization of the ASR by 0.6 mA-footshocks can be blocked by local microinjections of the SP-antagonists CP-96,345 (5 pmol-10 nmol) or CP-99,994 (0.5 nmol-100 nmol) into the PnC. Possible pathways relevant for the sensitization of the ASR are discussed.