Decoding vomeronasal activity through awake, behaving AOB imaging. - Summary: The sense of smell in the mouse largely occurs through the main olfactory system (MOS) which has the amazing ability to detect trace amounts of an infinite variety of volatile organic ligands. With this sensory power why does the mouse, and most terrestrial vertebrates, also have a vomeronasal organ (VNO)? Some think that since the VNO was lost during human evolution it is not important to study. First, we expect investigating the VNO will further advance our understanding of all types of chemosensation. The presence of a robust VNO across evolutionary diverse species indicates that taste and smell have functional blind spots that the VNO is filling. Understanding the VNO will enable us to know if humans are missing an animal superpower and can search for ways to augment our sensation, or if we evolved beyond its tether; perhaps losing this system partially accounts for our remarkable behavioral flexibility. Second, as an experimental model, the VNO provides unprecedented access to activate and study circuits critically important to humans. VNO mediated behavior has accelerated identification of neurons, circuits, and the brain network that underlies mouse social behavior and provides mechanistic insight to analogous circuits implicated in human social behavior. Anatomically, accessory olfaction has direct and privileged access to the social behavior brain network and genetic ablation of this system results in aberrant, dysfunctional social communication. Although humans are not thought to use olfaction to access social behavior circuits, the downstream network is critical and dysfunction leads to autism, ADHD, anxiety, or depression, but much is still unknown. Upon successful completion of this research, we will have created datasets quantitatively detailing and simultaneously bridging major unknowns about the accessory olfactory system: what it senses, metrics of neural activity encoding the environment, and properties of sensory coding in relationship to behavior. Together, our proposed work will investigate properties of VNO sensation that are expected to differ and complement the function of the MOS. Our method enables us to gain first insight about evoked sensory activity in relationship to the environment and behavior during dynamic natural behavior and is expected to generate new insight about chemosensory function.