Project Summary/Abstract Propensity toward relapse is a hallmark feature of addiction. Hence, understanding the cellular mechanisms responsible for relapse vulnerability represents an important focus of addiction research. One significant and long-lasting cellular adaptation observed in response to multiple drugs of abuse is downregulation of astroglial glutamate transporter GLT-1. However, relatively very little is known about how drug self-administration affects astrocytes beyond GLT-1 expression, or how astrocytes may contribute to mechanisms of drug seeking. Results collected during the preceding K99/R00 award indicate that restored expression of GLT-1 is pivotal to the mechanism of action of multiple compounds that reduce behavioral measures of relapse in the rat self- administration and reinstatement model of addiction. Preliminary data also indicate that downregulation of GLT-1 by cocaine is accompanied by reduced expression of glial fibrillary acidic protein (GFAP) and a retraction of astrocytes in the nucleus accumbens core. Astrocyte retraction is characterized by decreased surface area, volume, and decreased synaptic contacts. This finding represents a heretofore-unappreciated fundamental consequence of cocaine use on astrocyte cell biology. Thus, decreased GLT-1 expression is a component of larger-scale adaptions in astrocyte biology that occur following chronic cocaine use. These findings have led to the hypothesis that astrocyte retraction in the nucleus accumbens of cocaine-withdrawn rats contributes to synaptic adaptations that drive cocaine seeking. In order to test this hypothesis, the specific goals of this proposal are: (1) to determine when during the addiction cycle the morphological effects on astrocytes are induced (2) to determine the functional relationship between astrocyte retraction and synaptic adaptations believed to underlie cocaine seeking, and (3) to determine the relationship between astrocyte retraction and drug seeking after cessation of drug use. These questions will be addressed by combining rat cocaine self-administration with behavioral measures of cocaine seeking, high-resolution imaging of fluorescently labeled astrocytes, and whole cell patch-clamp electrophysiology. These studies will provide novel insight into how cocaine-dependent adaptations in astrocyte dynamics contribute to the cellular and behavioral pathologies characteristic of psychostimulant addiction. These studies will also provide important information toward the translational potential of astrocytes as a pharmacotherapeutic target for substance use disorders.