Little is known about the consequences of altered endocannabinoid signalling in adolescence. We hypothesized that CB1 receptor antagonism (AM251, 1 mg/kg) and stress exposures (1 h confinement stress) in adolescence (daily, postnatal days 30–44) would interact to increase neuroendocrine stress responses and anxiety when investigated a minimum of 24 h after drug and stress treatments; these treatment effects were independent of each other. Changes in homecage behaviour and in weight gain confirmed that both males and females were sensitive to the treatments. Nevertheless, in males, repeated AM251 administration was without effect on any of the measures investigated in days post-treatment. Males had reduced corticosterone release to the repeated stress and had increased GAD67 expression in the ventral hippocampus under baseline conditions. In females, AM251 also reduced weight gain and increased stereotypic behaviours in the homecage; these same females showed increased sociality, reduced CB1 receptor expression in the dorsal hippocampus, and increased GAD67 expression in the prefrontal cortex. Further, females exposed to repeated stress had enhanced recovery to baseline corticosterone concentrations after stress. The inclusion of a non-injected comparison group also revealed stress of injection effects in both sexes that otherwise would have been masked. Together, the findings demonstrate effects of CB1 receptor antagonism and stress that were more evident in females than males, suggesting that females may be more vulnerable to the consequences of disrupted endocannabinoid signalling during adolescence.