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Nitric oxide (
•
NO) is a ubiquitous signaling molecule that participates in neuromolecular phenomena associated with memory formation as well as in excitotoxicity. In the hippocampus, neuronal •NO production is coupled to the activation of the NMDA-type of glutamate receptor. More recently, Cytochrome c oxidase has emerged as a novel target for •NO, which competes with O
2
for binding to this mitochondrial complex. This reaction establishes •NO not only as a regulator of cellular metabolism but possibly also as a regulator of mitochondrial production of reactive oxygen species which participate in cellular signaling. A major gap in the understanding of •NO bioactivity, namely, in the hippocampus, has been the lack of knowledge of its concentration dynamics. Here, we present a detailed description of the simultaneous recording of •NO and O
2
concentration dynamics in rat hippocampal slices. Carbon fiber microelectrodes are fabricated and applied for real-time measurements of both gases in a system close to
in vivo
models. This approach allows for a better understanding of the current paradigm by which an intricate interplay between •NO and O
2
regulates cellular respiration.