A heightened sensitivity to unpredictable aversiveness is a key component of several anxiety disorders. Neuroimaging studies of unpredictable aversiveness have shown that the anterior region of the insula cortex (AIC) plays a central role in the anticipation of unpredictable aversiveness. The present study extended these findings by examining the role of the AIC in temporal unpredictability (i.e. not knowing when the stimulus will occur), a particularly critical aspect of unpredictability as it increases contextual anxiety and vigilance, given that the danger could happen ‘at any time’. Nineteen healthy participants underwent functional MRI while anticipating either temporally unpredictable or predictable aversive (or neutral) images. Participants showed greater right AIC activation while anticipating unpredictable relative to predictable aversive images. In addition, activation in this region was correlated positively with self-reported individual differences in a key facet of intolerance of uncertainty (inhibitory behavior). Taken together, the present study suggests that the AIC plays an important role in the anticipation of temporally unpredictable aversiveness and may mediate key deficits in anxiety disorders.