1H- and 31P-nuclear magnetic resonance studies of l-lactate transport in isolated muscle fibers from the spiny lobster Panulirus argus

Abstract

Proton (1H) and phosphorus (31P) nuclear magnetic resonance (NMR) spectroscopy were used to investigate the mode of transport of l-lactate across the plasma membranes of the abdominal extensor muscles of the spiny lobster Panulirus argus. Individual fibers or bundles of 2­3 fibers were superfused in a dual-tuned (1H, 31P) microsolenoid NMR probe. 1H-NMR spectra were diffusion-weighted, which eliminated the signal contribution of the fast-flowing extracellular lactate but retained that of intracellular lactate. Well-resolved intracellular lactate signals could be acquired at 15 s intervals, permitting estimation of initial velocities (Vi) of influx and efflux during loading/unloading of muscle fibers. 31P-NMR spectra were acquired to assess cellular energy status and intracellular pH. Transport results showed that Vi values for influx and efflux were a linear function of total lactate concentration, displaying no saturation effects. The rate of lactate influx was enhanced by increasing the concentration of the free acid by altering the superfusate pH. Vi values for influx and efflux of d- and l-lactate were identical. Finally, traditional inhibitors of monocarboxylate and/or anion transport had no effect on influx/efflux of lactate from these cells. The above results strongly suggest that the primary mode of lactate transport is by passive diffusion. These cells appear to lack a monocarboxylate transporter, which may be related to the apparent absence of organ-specific compartmentation of lactate metabolism in crustaceans.