A posterior insula to lateral amygdala pathway transmits US-offset information with a limited role in fear learning

During fear learning, associations between a sensory cue (conditioned stimulus, CS) and an aversive stimulus (unconditioned stimulus, US) are formed in specific brain circuits. The lateral amygdala (LA) is involved in CS-US integration; however, US pathways to the LA remain understudied. Here, we investigated whether the posterior insular cortex (pInsCx), a hub for aversive state signaling, transmits US information to the LA during fear learning. We find that the pInsCx makes a robust, glutamatergic projection specifically targeting the anterior LA. In vivo Ca²⁺ imaging reveals that neurons in the pInsCx and anterior LA display US-onset and US-offset responses; imaging combined with axon silencing shows that the pInsCx selectively transmits US-offset information to the anterior LA. Optogenetic silencing, however, does not show a role for US-driven activity in the anterior LA or its pInsCx afferents in fear memory formation. Thus, we describe a cortical projection that carries US-offset information to the amygdala with a limited role in fear learning.