We have examined the effect of lesions of 5‐hydroxytryptamine (5‐HT) neurons, produced by p‐chloroamphetamine (p‐CA; 2 times 10 mg kg−1), and the influence of flumazenil (Ro 15–1788, 10 mg kg−1), a benzodiazepine receptor antagonist, on the anxiolytic‐like activity of CP 94253 (5‐propoxy‐3‐(1,2,3,6‐tetrahydro‐4‐pyridinyl)‐1‐H‐pyrrolo[3,2‐b]pyridine), a 5‐HT1Breceptor agonist, SB 216641 (N‐[3‐[3‐(dimethylamino)ethoxy]‐4‐methoxyphenyl]‐2′‐methyl‐4′‐(5‐methyl‐1,2,4‐oxadiazol‐3‐yl)‐[1,1′‐biphenyl]‐4‐carboxamide), a 5‐HT1Breceptor antagonist, and GR 127935 (N‐[4‐methoxy‐3‐(4‐methyl‐l‐piperazinyl)phenyl]‐2′‐methyl‐4′‐(5‐methyl‐1,2,4‐oxadiazol‐3‐yl)‐1, 1′‐biphenyl‐4‐carboxamide), a 5‐HT1B/1Dreceptor antagonist, in the Vogel conflict drinking test in rats. Diazepam was used as a reference compound. CP 94253 (2.5 mg kg−1), SB 216641 (2.5 mg kg−1), GR 127935 (10 mg kg−1) and diazepam (5 mg kg−1) significantly increased the number of shocks accepted during experimental sessions in the conflict drinking test in vehicle‐ and p‐CA‐pretreated rats. Flumazenil did not change the anxiolytic‐like effect of CP 94253 (2.5 mg kg−1), but wholly blocked the anxiolytic‐like effects of SB 216641 (2.5 mg kg−1), GR 127935 (10 mg kg−1) and diazepam (5 mg kg−1). p‐CA and flumazenil alone were inactive in the conflict drinking test. The results suggested that the anxiolytic‐like effect of the 5‐HT1Breceptor ligands CP 94253, SB 216641 and GR 127935 was possibly linked to the postsynaptic 5‐HT1Breceptors or/and 5‐HT1Bheteroreceptors. The results suggested also that benzodiazepine receptors were indirectly involved in the effects of SB 216641 and GR 127935 (but not of CP 94253), which might have been due to a possible interaction between the 5‐HT and the GABA/benzodiazepine systems.