Genetic Disruption of System xc-Mediated Glutamate Release from Astrocytes Increases Negative-Outcome Behaviors While Preserving Basic Brain Function in Rat

Abstract

The importance of neuronal glutamate to synaptic transmission throughout the brain illustrates the immense therapeutic potential and safety risks of targeting this system. Astrocytes also release glutamate, the clinical relevance of which is unknown since the range of brain functions reliant on signaling from these cells hasn’t been fully established. Here, we investigated system xc- (Sxc), which is a glutamate release mechanism with anin vivorodent expression pattern that is restricted to astrocytes. Since most animals do not express Sxc, we first compared the expression and sequence of the obligatory Sxc subunit xCT among major classes of vertebrate species. We found xCT to be ubiquitously expressed and under significant negative selective pressure. Hence, Sxc likely confers important advantages to vertebrate brain function that may promote biological fitness. Next, we assessed brain function in male genetically-modified rats (MSxc) created to eliminate Sxc activity. Unlike other glutamatergic mechanisms, eliminating Sxc activity was not lethal and didn’t alter growth patterns, telemetry measures of basic health, locomotor activity, or behaviors reliant on simple learning. However, MSxc rats exhibited deficits in tasks used to assess cognitive behavioral control. In Pavlovian conditioned approach, MSxc rats approached a food-predicted cue more frequently than WT rats, even when this response was punished. In attentional set-shifting, MSxc rats displayed cognitive inflexibility due to an increased frequency of perseverative errors. MSxc rats also displayed heightened cocaine-primed drug seeking. Hence, a loss of Sxc-activity appears to weaken control over non-reinforced or negative-outcome behaviors without altering basic brain function.Significance Statement:Glutamate is essential to synaptic activity throughout the brain, which illustrates immense therapeutic potential and risk. Notably, glutamatergic mechanisms are expressed by most types of brain cells. Hence, glutamate likely encodes multiple forms of intercellular signaling. Here, we hypothesized that the selective manipulation of astrocyte to neuron signaling would alter cognition without producing widespread brain impairments. First, we eliminated activity of the astrocytic glutamate release mechanism, system xc (Sxc), in rat. This impaired cognitive flexibility and increased expression of perseverative, maladaptive behaviors. Notably, eliminating Sxc activity did not alter metrics of health or non-cognitive brain function. These data add to recent evidence that the brain expresses cognition-specific molecular mechanisms that could lead to highly precise, safe medications for impaired cognition.

Sex- and age-dependent contribution of System xc– to cognitive, sensory, and social behaviors revealed by comprehensive behavioral analyses of System xc– null mice

“…Notably, all mice, irrespective of their genotype, sex, or age, exhibited a positive NOR discrimination index ( Figures 7B, D ), indicating that recognition memory of Sx c – null mice remained intact. These results are consistent with a previous study conducted in rats with a genetic disruption of Sx c – ( Hess et al, 2023 ). However, our investigation using the Barnes maze revealed disruptions in certain aspects of spatial memory ( Figure 8 ).…”