Hepatocyte heterogeneity in glutamate metabolism and bidirectional transport in perfused rat liver

1 The metabolic fate of infused [1‐ 14 C]glutamate was studied in perfused rat liver. The 14 C label taken up by the liver was recovered to 85 ± 2% as 14 CO 2 and [ 14 C]glutamine. Whereas 14 CO 2 production accounted for about 70% of the [1‐ 14 C]glutamate taken up under conditions of low endogenous rates of glutamine synthesis, stepwise stimulation of glutamine synthesis by NH 4 Cl increased 14 C incorporation into glutamine at the expense of 14 CO 2 production. Extrapolation to maximal rates of hepatic glutamine synthesis yielded an about 100% utilization of vascular glutamate taken up by the liver for glutamine synthesis. This was observed in both, antegrade and retrograde perfusions and suggests an almost exclusive uptake of glutamate into perivenous glutamine‐synthetase‐containing hepatocytes. 2 Glutamate was simultaneously taken up and released from perfused rat liver. At a near‐physiological influent glutamate concentration (0.1 mM), the rates of unidirectional glutamate influx and efflux were similar (about 100 and 120 nmol g −1 min −1 , respectively). 3 During infusion of [1‐ 14 C]oxoglutarate (50 μM), addition of glutamate (2 mM) did not affect hepatic uptake of [1‐ 14 C]oxoglutarate. However, it increased labeled glutamate release from the liver about 10‐fold (from 9 ± 2 to 86 ± 20 nmol g −1 min −1 ; n = 4), whereas 14 CO 2 production from labeled oxoglutarate decreased by about 40%. This suggests not only different mechanisms of oxoglutarate and glutamate transport across the plasma membrane, but also points to a glutamate/glutamate exchange. 4 Oxoglutarate was recently shown to be taken up almost exclusively by perivenous glutamine‐synthetase‐containing hepatocytes [Stoll, B & Häussinger, D. (1989) Eur. J. Biochem. 181 , 709–716] and [1‐ 14 C]oxoglutarate (9 μM) was used to label selectively the intracellular glutamate pool in this perivenous cell population. The specific radioactivity of this intracellular (perivenous) glutamate pool was assessed by measuring the specific radioactivity of newly synthesized glutamine which is continuously released from these cells into the perfusate. Comparison of the specific radioactivities of glutamine and glutamate released from perivenous cells indicates that about 60% of total glutamate release from the liver is derived from the perivenous glutamine‐synthetase‐containing cell population. Following addition of unlabeled glutamate (0.1 mM), unidirectional glutamate efflux from perivenous cells increased from about 30 to 80 nmol g −1 min −1 , whereas glutamate efflux from non‐perivenous (presumably periportal) hepatocytes remained largely unaltered (i.e. 20 – 30 nmol g −1 min −1 ). 5 It is concluded that, in the intact liver, vascular glutamate is almost exclusively taken up by the small perivenous hepatocyte population containing glutamine synthetase. Glutamate release from the liver, which occurs simultaneously and can be stimulated by extracellular glutamate, must also largely (i.e. about 80%) be ascribed to this small cell population This suggests that hepatic glutamate uptake occurs, at least in part via a glutamate/ glutamate exchange mechanism, which is restricted to the perivenous cell population. The findings extend the concept of a structural/functional acinar organization of hepatic nitrogen and amino acid metabolism to bidirectional glutamate fluxes across the plasma membrane in line with recent suggestions [Häussinger, D & Gerok, W. (1983) Eur. J. Biochem. 136 , 421–425].