Project Summary Cellular mechanisms mediating the transition from a state of general anesthesia to an awake state are not understood. Patients emerge from anesthesia passively without the use of mechanistically targeted interventions, creating an unpredictable clinical outcome marked by behavioral phenomena like emergence agitation and delirium. Many anesthetics act either directly or indirectly to change excitatory/inhibitory balance in the brain, highlighting the importance of understanding inhibitory networks in emergence. My research program focuses on the effects of general anesthesia on inhibitory plasticity during the transition from anesthetized to awake state in mice. My prior work examines neuroligin-2, a central organizer of the inhibitory synapse, using cell-type and circuit specific manipulations. Neuroligin-2 is a cell adhesion protein that acts as a scaffold to regulate general inhibitory synaptic function and is recently implicated as an independent regulator of intracellular signaling and disease. I demonstrated that neuroligin-2 manipulation modulates agitation and related behaviors in mouse models. My established expertise in stress-induced inhibitory synaptic plasticity provides a strong foundation for the following three complementary research areas investigating general anesthesia emergence. (1) Inhibitory cell plasticity in emergence from anesthesia: Research area 1 will focus on effects of general anesthesia emergence on inhibitory cell plasticity, starting with an investigation of key inhibitory postsynaptic genes, like neuroligin-2, using targeted knockdown and electrophysiology studies. Single cell sequencing transcriptomic investigations will characterize all cell types and synaptic constituents modified by emergence. (2) Brain-wide cell type-specific circuit activity in emergence: Research area 2 will investigate whole brain circuit changes induced by general anesthesia emergence using activity mapping and light sheet microscopy at single cell resolution, along with recordings of neuronal activity using genetically encoded optical sensors expressed in anesthesia-regulated circuits. (3) Preclinical models for emergence delirium across the lifespan: Research area 3 will develop and validate preclinical rodent models of emergence delirium using machine learning approaches, in order to study vulnerability to anesthesia-induced delirium across the lifespan. Together, these three projects form an overarching research program to understand mechanisms of emergence from the inhibitory cellular to circuit to behavioral levels of analysis, providing a holistic view of emergence. We must understand the mechanisms of anesthetic emergence across all levels in order to design interventions to bring about safer and predictable emergence.