The neural substrate underlying the aversive effects induced by ethanol abstinence is still unclear. One candidate for such effects is the dorsal periaqueductal gray (dPAG), a core structure of the brain aversion system. The main aim of this study is to examine the role of the dPAG as a possible locus of the aversive effects following abrupt alcohol withdrawal. To this end, rats were subjected to an oral ethanol self-administration procedure, in which animals were offered 6-8% (v/v) ethanol solution for a period of 21 days followed by an abrupt discontinuation of the treatment on the two subsequent days. Control animals received control dietary fluid for similar periods of time. The effects of ethanol withdrawal were examined in the elevated plus-maze (EPM) (Exp. I), on the prepulse inhibition of startle to loud sounds (Exp. II) and on the freezing and escape responses induced by electrical stimulation of the dPAG (Exp. III). In Experiment III, rats were implanted with an electrode aimed at the dPAG and the number and duration of ultrasonic vocalizations (USVs) were also recorded in the rats that received dPAG stimulation at freezing and escape thresholds. Data obtained showed that ethanol withdrawal elicited significant "anxiety-like" behaviors, as revealed by the decrease in the number of entries into and time spent onto the open arms of the EPM. Startle reflex and prepulse inhibition remained unchanged in withdrawn animals. In addition, discontinuation from the chronic ethanol regimen caused a reduction in the stimulation thresholds for freezing and escape and in the number and duration of USVs. Together, these effects have been interpreted in the frame of a high fear state elicited by activation of the dPAG. These findings are indicative that ethanol withdrawal sensitizes the substrates of fear at the level of this midbrain structure.