Central CO₂ chemosensitive neurons in the caudal solitary complex (cSC) are stimulated not only by hypercapnic acidosis (HA), but by hyperoxia as well. While a cellular mechanism for the CO₂ response has yet to be isolated, previous data show that a redox-sensitive mechanism underlies neuronal excitability to hyperoxia. However, it remains unknown how changes in pO₂ affect the production of reactive oxygen and nitrogen species (RONS) in the cSC that can lead to increased cellular excitability and with larger doses, cellular dysfunction and death. To this end, we used fluorescence microscopy in real time to determine how normobaric hyperoxia increases the production of key RONS in the cSC. Because neurons in the region are CO₂ sensitive, we also examined the potential effects of CO₂ narcosis, used during euthanasia prior to brain slice harvesting, on RONS production. Our findings show that normobaric hyperoxia (0.4 0.95 ATA O₂) increases the fluorescence rates of fluorogenic dyes specific to both superoxide and nitric oxide. Interestingly, different results were seen for superoxide fluorescence when CO₂ narcosis was used during euthanasia, suggesting long-lasting changes in superoxide production and/or antioxidant activity subsequent to CO₂ narcosis prior to brain slicing. Further research needs to distinguish whether the increased levels of RONS reported here are merely increases in oxidative and nitrosative signaling or, alternatively, evidence of redox and nitrosative stress.