Studying rat brain neurochemistry using nanoprobe NMR spectroscopy: a metabonomics approach

In the present experiments, in vivo microdialysis techniques together with nanoprobe NMR spectroscopy were used to evaluate the neurochemical environment of the rat frontal cortex. Metabonomics techniques of data reduction and pattern recognition were used to examine whether collected neurochemicals were sensitive to tetrodotoxin (TTX), a neurotoxin that when infused into discrete brain regions can help distinguish between the neuronal versus glial origin of neurochemicals in cerebrospinal fluid microdialysate. (1)H NMR spectra recorded on samples collected from the rat frontal cortex before and after an intracortical TTX infusion (10 microM for 60 min) were subjected to multivariate statistical analysis. Glutamate, isoleucine, valine, alanine, and alpha- and beta-hydroxybutyrate were found to have decreased concentrations after the addition of TTX, suggesting that their release is likely from cortical neurons. In contrast, lactate, formate, acetate, glucose, creatinine, pyruvate, and other neurochemicals remained unchanged following local application of TTX. The present findings extend our previous work combining the analytical technology of small-volume nanoprobe NMR spectroscopy with in vivo microdialysis in freely moving animals and show that it is possible to apply metabonomics methodology to this important class of biofluid to monitor changes in neurochemical composition of the rat brain.