Psychopharmacology (1998) 135 : 342352 © Springer-Verlag 1998 ORIGINAL INVESTIGATION B.O. Ward · D.N. Stephens Sensitisation of withdrawal signs following repeated withdrawal from a benzodiazepine: differences between measures of anxiety and seizure sensitivity Received : 13 April 1997 / Final version : 14 August 1997 Abstract Three experiments examined the e¤ect of either withdrawal from diazepam, or repeated treat- ment with the convulsant, pentylenetetrazol (PTZ), on behaviour and seizure threshold. The behaviours mea- sured were on the elevated plus maze and in the four- plate test; seizure threshold was measured as dose of PTZ infused via the tail vein to the Þrst clonic twitch. In experiment 1, we examined the e¤ect of either sin- gle or repeated withdrawal from diazepam using a pro- cedure in which the drug was administered SC in a slow release depot. Three cycles of withdrawal from diazepam were compared to a single withdrawal expe- rience. A single withdrawal from diazepam following chronic treatment gave rise, 72 h following the last dos- ing, to behavioural changes, suggestive of anxiety, in both tests, but did not result in a reduced convulsant threshold. In contrast, repeated withdrawal resulted in a reduction in sensitivity in several measures of anxi- ety, but sensitised the mice to the convulsive e¤ects of the PTZ. The unexpected failure to Þnd an increased sensitivity to a convulsive agent following a single with- drawal from SC diazepam was examined in experiment 2. The seizure threshold following a single withdrawal of mice which had received diazepam chronically IP in aqueous vehicle was signiÞcantly reduced relative to vehicle-treated controls, whereas that of animals receiv- ing the same dose SC in oil, was not. It is argued that the di¤erence may arise from the animals treated repeatedly with IP diazepam unintentionally experi- encing repeated withdrawal, since the half-life of the drug by this route is short. In experiment 3, repeated sub-convulsant PTZ treatment reduced the convulsant threshold (the dose of PTZ required to give rise to the Þrst clonic twitch), but had no signiÞcant e¤ect on the behavioural measures of anxiety compared to a single dose of PTZ or vehicle controls. The results suggest that repeated withdrawal from chronic treatments with diazepam sensitises mice to convulsant stimuli in a manner resembling the e¤ects of repeated administra- tion of sub-convulsant doses of PTZ, but that neither repeated PTZ nor repeated diazepam withdrawal results in increased sensitivity to anxiogenic stimuli; rather, repeated withdrawal from diazepam may reduce the susceptibility of mice to behavioural measures of anxiety. Key words Conßict · Diazepam · Pentylenetetrazole · Mice · Plus maze · Four plate · Repeated withdrawal · Kindling · Dependence · Anxiety Introduction Long-term treatment with sedative-hypnotic drugs including alcohol, barbiturates and benzodiazepines may lead to dependence, manifested as a syndrome of abstinence signs and symptoms following drug with- drawal. In the case of alcohol, a number of clinical and experimental reports suggest that previous experience of withdrawal increases the severity of subsequent with- drawal episodes. Thus patients undergoing alcohol withdrawal are more likely to experience seizures if they have undergone previous episodes of detoxi Þcation (Mendelson et al. 1966; Baker and Cannon 1979; Brown et al. 1988; Lechtenberg and Worner 1991; Booth and Blow 1993). Analogous Þndings have also been reported from animal experiments which showed that the severity of the withdrawal intensity increased following several withdrawal episodes (Branchey et al. 1971; Walker and Zornetzer 1974; Baker and Cannon 1979; Clemmesen and Hemmingsen 1984; Maier and Pohoreky 1989; Becker and Hale 1993; Kokka et al. B.O. Ward · D.N. Stephens (*) Laboratory of Experimental Psychology, University of Sussex, Falmer, Brighton BN1 9QG, UK Fax (+44) 1273-678611, e-mail:dns@biols.susx.ac.uk