Anxiety disorders, such as post-traumatic stress disorder (PTSD), are thought to occur by dysfunction in the fear and anxiety-related brain circuit, however, the exact mechanisms remain unknown. Recent human studies have shown that the right anterior insular cortex (aIC) activity is positively correlated with the severity of PTSD symptoms. Understanding the role of the aIC in fear and anxiety may provide insights into the etiology of anxiety disorders. We used a modified shock-probe defensive burying behavioral test, which utilizes the natural propensity of rodents to bury potentially dangerous objects, to test the role of aIC in fear. Mice exposed to restraint stress exhibited burying of the restrainer-resembling object, indicative of defensive behavior. Electrolytic ablation of the aIC significantly diminished this defensive burying behavior, suggesting the involvement of the aIC. Single-unit recording of pyramidal neurons in the aIC showed that a proportion of neurons which increased activity in the presence of a restrainer-resembling object was significantly correlated with the defensive burying behavior. This correlation was only present in mice exposed to restraint stress. These results suggest that altered neuronal representation in the aIC may regulate fear and anxiety after exposure to a traumatic event. Overall, our result demonstrates that the aIC mediates fear and anxiety and that it could be a potential target for treating anxiety disorders.