ABSTRACT: Oxidative stress figures prominently in retinal diseases, including diabetic retinopathy, and glaucoma. Ligands for ?1R, a unique transmembrane protein localized to the endoplasmic reticulum, mitochondria, and nuclear and plasma membranes, have profound retinal neuroprotective properties in vitro and in vivo. Studies to determine the mechanism of ?1R-mediated retinal neuroprotection have focused mainly on neurons. Little is known about the effects of ?1R on Müller cell function, yet these radial glial cells are essential for homeostatic support of the retina. Here we investigated whether ?1R mediates the oxidative stress response of Müller cells using wild-type (WT) and ?1R-knockout (?1RKO) mice. We observed increased endogenous reactive oxygen species (ROS) levels in ?1RKO Müller cells compared to WT, which was accompanied by decreased expression of Sod1, catalase, Nqo1, Hmox1, Gstm6, and Gpx1. The protein levels of SOD1, CAT, NQO1, and GPX1 were also significantly decreased. The genes encoding these antioxidants contain an antioxidant response element (ARE), which under stress is activated by NRF2, a transcription factor that typically resides in the cytoplasm bound by KEAP1. In the ?1RKO Müller cells Nrf2 expression was decreased significantly at the gene (and protein) level, whereas Keap1 gene (and protein) levels were markedly increased. NRF2-ARE binding affinity was decreased markedly in ?1RKO Müller cells. We investigated system xc(-), the cystine-glutamate exchanger important for synthesis of glutathione (GSH), and observed decreased function in ?1RKO Müller cells compared to WT as well as decreased GSH and GSH/GSSG ratios. This was accompanied by decreased gene and protein levels of xCT, the unique component of system xc(-). We conclude that Müller glial cells lacking ?1R manifest elevated ROS, perturbation of antioxidant balance, suppression of NRF2 signaling, and impaired function of system xc(-). The data suggest that the oxidative stress-mediating function of retinal Müller glial cells may be compromised in the absence of ?1R. The neuroprotective role of ?1R may be linked directly to the oxidative stress-mediating properties of supportive glial cells.