Homonuclear 1H double-resonance difference spectroscopy of the rat brain in vivo

We have used 1H homonuclear double-resonance difference spectroscopy at 360.13 MHz to resolve specific metabolite resonances in the brains of intact rats. Metabolite resonances resolved include previously obscured proton resonances of alanine, gamma-aminobutyric acid (GABA), glutamate, and taurine. The gamma-aminobutyric acid alpha- and gamma-CH2 proton resonances were observed in the living rat in the difference spectrum obtained upon irradiation of the beta-CH2 proton resonance at 1.91 ppm. A 3-fold increase in the intensity of the alpha- and gamma-CH2 resonances of gamma-aminobutyric acid was observed 30 min after death. The alpha-CH and gamma-CH2 resonances of glutamate were also resolved in vivo by selective irradiation of the beta-CH2 protons to which they are spin-coupled. In addition, this technique was used to observe the beta-CH3 protons of lactate through the intact scalp of a rat. Large lipid signals arising from scalp tissue were eliminated in the difference spectrum, revealing the lactate beta-CH3 resonance.