31P-magnetic resonance spectroscopy of the rabbit masseter muscle

Dynamic biochemical changes in the masseter muscle were studied in 14 New Zealand adult male rabbits by 31P-nuclear magnetic resonance (NMR) spectroscopy. NMR spectra were obtained during rest and electrical stimulation of the muscle in the anaesthetized animal at 33 recording sessions. Electrical stimulation was applied by a pair of copper wires placed separately with hypodermic needles into the muscle. NMR spectra were acquired with a 2 x 3 cm, double-turn, copper transmit/receive coil. Sixteen spectra were averaged over 30 s to obtain averaged spectra continuously during a 30-min recording. The spectra were processed automatically using a non-linear 'least-squares' fitting program on the spectrometer. A Lorentzian line shape was assumed for the peaks, and values of peak height, area and chemical shifts were generated. Each averaged spectrum consisted of five peaks: inorganic phosphate (Pi), creatine phosphate (PCr), and three peaks related to ATP. Data were analysed as to absolute changes in Pi and PCr, in the ratio of Pi/PCr, and the shift of Pi to PCr to estimate pH. Several protocols were used in which ranges of frequency, intensity and duration of electrical stimulation were tested. The protocol for detailed studies involved stimulating the muscle twice at 5 Hz for 3 min separated by a 3-min rest period, then stimulating twice at 50 Hz for 3 min separated by a rest period. During contraction of the muscle, there was a significant increase in the Pi/PCr ratio (p < 0.05) as compared to the resting level. The ratio reached a plateau over a 3-min contraction using 5-Hz stimulation, then increased significantly more with the 50-Hz stimulation but decayed during the 3 min. Sustained stimulation with 50 Hz for 15-45 min evoked an initial sharp change in Pi/PCr, which then reached a steady plateau that remained over the entire stimulation. These findings indicate that the rabbit masseter muscle is relatively fatigue resistant in maintaining a steady-state equilibrium in the relation of Pi to PCr.