Angiotensin II Type 2 Receptor-Expressing Neurons in the Central Amygdala Influence Fear-Related Behavior

Background: The renin-angiotensin system has been implicated in posttraumatic stress disorder; however, the mechanisms responsible for this connection and the therapeutic potential of targeting the renin-angiotensin system in posttraumatic stress disorder remain unknown. Using an angiotensin receptor bacterial artificial chromosome (BAC) and enhanced green fluorescent protein (eGFP) reporter mouse, combined with neuroanatomical, pharmacological, and behavioral approaches, we examined the role of angiotensin II type 2 receptor (AT₂R) in fear-related behavior.

Methods: Dual immunohistochemistry with retrograde labeling was used to characterize AT₂R-eGFP⁺ cells in the amygdala of the AT₂R-eGFP-BAC reporter mouse. Pavlovian fear conditioning and behavioral pharmacological analyses were used to demonstrate the effects of AT₂R activation on fear memory in male C57BL/6 mice.

Results: AT₂R-eGFP⁺ neurons in the amygdala were predominantly expressed in the medial amygdala and the medial division of the central amygdala (CeM), with little AT₂R-eGFP expression in the basolateral amygdala or lateral division of the central amygdala. Characterization of AT₂R-eGFP⁺ neurons in the CeM demonstrated distinct localization to gamma-aminobutyric acidergic projection neurons. Mice receiving acute intra-central amygdala injections of the selective AT₂R agonist compound 21 prior to tests for cued or contextual fear expression displayed less freezing. Retrograde labeling of AT₂R-eGFP⁺ neurons projecting to the periaqueductal gray revealed AT₂R-eGFP⁺ neuronal projections from the CeM to the periaqueductal gray, a key brain structure mediating fear-related freezing.

Conclusions: These findings suggest that CeM AT₂R-expressing neurons can modulate central amygdala outputs that play a role in fear expression, providing new evidence for a novel angiotensinergic circuit in the regulation of fear.