Rewinding the tape: selection of algae adapted to high CO2 at current and pleistocene levels of CO2

Selective history is thought to constrain the extent and direction of future adaptation by limiting access to genotypes that are advantageous in a novel environment. Populations of Chlamydomonas previously selected at high CO2 were either backselected at ambient levels of CO2, or selected at levels of CO2 that last occurred during glaciation in the Pleistocene. There was no effect of selective history on adaptation to either level of CO2, and the high CO2 phenotypes were evolutionarily reversible such that fitness in ambient CO2 returned to values seen in controls. CO2 uptake affinity improved relative to the ancestor in both ambient and glacial CO2, although wild-type regulation of CO2 uptake, which deteriorated during previous selection at high CO2, was not restored by selection at lower levels of CO2. Trade-offs in both CO2 uptake affinity and growth were seen after selection at any given level of CO2. Adaptation to ambient and glacial-era levels of CO2 produced a range of phenotypes, suggesting that chance rather than selective history contributes to the divergence of replicate populations in this system.     

Short-Term Regulation of Crassulacean Acid Metabolism Activity in a Tropical Hemiepiphyte,Clusia uvitana

Diel courses of net CO2 exchange of leaves were studied in Clusia uvitana (Clusiaceae), a tropical Crassulacean acid metabolism (CAM) hemiepiphyte, growing in the crown of a 47-m tall kapok tree on Barro Colorado Island, Panama. Measurements on days without precipitation showed that net uptake of atmospheric CO2 occurred at night, a feature of CAM, as well as in the early morning and late afternoon. During 36 h of almost continuous rainfall, nocturnal net CO2 uptake was abolished and the diel pattern of net CO2 exchange became similar to that of a C3 plant. Exposing well-watered, potted plants of Clusia in the laboratory to temperatures and photosynthetic photon flux densities similar to those during the tropical rainstorm also abolished nocturnal net CO2 uptake. In contrast, Kalanchoe pinnata (Crassulaceae), an obligate CAM plant, still showed net CO2 dark fixation following the same low-light and moderate-temperature conditions, albeit at decreased rates. During these 12-h photoperiods, titratable acidity in Clusia increased slightly above its high level measured at the end of the previous dark period, whereas in Kalanchoe, the acid content decreased by about 40%. A survey among outer canopy leaves of Clusia on Barro Colorado Island showed that leaves that exhibited little or no nocturnal acidification maintained high levels of H+ at dawn and dusk. Progressively lower levels of H+ at dusk were accompanied by progressively higher nocturnal increases in H+. The data suggest that in C. uvitana the rapid switching between CAM- and C3-type carbon fixation that may occur within 24 h in response to environmental changes is controlled by the acidity status of the leaves in the light. Nocturnal CO2 fixation is enhanced by conditions that decrease the organic acid content during the light period.     

Responses of global plant diversity capacity to changes in carbon dioxide concentration and climate

We model plant species diversity globally by country to show that future plant diversity capacity has a strong dependence on changing climate and carbon dioxide concentration. CO2 increase, through its impact on net primary production and warming is predicted to increase regional diversity capacity, while warming with constant CO2 leads to decreases in diversity capacity. Increased CO2 concentrations are unlikely to counter projected extinctions of endemic species, shown in earlier studies to be more strongly dependent on changing land use patterns than climate per se. Model predictions were tested against (1) contemporary observations of tree species diversity in different biomes, (2) an independent global map of contemporary species diversity and (3) time sequences of plant naturalisation for different locations. Good agreements between model, observations and naturalisation patterns support the suggestion that future diversity capacity increases are likely to be filled from a 'cosmopolitan weed pool' for which migration appears to be an insignificant barrier.     

Hepatocyte heterogeneity in glutamate uptake by isolated perfused rat liver

Glutamate is simultaneously taken up and released by perfused rat liver, as shown by 14CO2 production from [1-14C]glutamate in the presence of a net glutamate release by the liver, turning to a net glutamate uptake at portal glutamate concentrations above 0.3 mM. 14CO2 production from portal [1-14C]glutamate is decreased by about 60% in the presence of ammonium ions. This effect is not observed during inhibition of glutamine synthetase by methionine sulfoximine. 14CO2 production from [1-14C]glutamate is not influenced by glutamine. Also, when glutamate accumulates intracellularly during the metabolism of glutamine (added at high concentrations, 5 mM), 14CO2 production from [1-14C]glutamate is not affected. If labeled glutamate is generated intracellularly from added [U-14C]proline, stimulation of glutamine synthesis by ammonium ions did not affect 14CO2 production from [U-14C]proline. After induction of a perivenous liver cell necrosis by CCL4, i.e. conditions associated with an almost complete loss of perivenous glutamine synthesis but no effect on periportal urea synthesis, 14CO2 production from [1-14C]glutamate is decreased by about 70%. The results are explained by hepatocyte heterogeneity in glutamate metabolism and indicate a predominant uptake of glutamate (that reaches the liver by the vena portae) by the small perivenous population of glutamine-synthesizing hepatocytes, whereas glutamate production from glutamine or proline is predominantly periportal. In view of the size of the glutamine synthetase-containing hepatocyte pool [Gebhardt, R. and Mecke, D. (1983) EMBO J. 2, 567-570], glutamate transport capacity of these hepatocytes would be about 20-fold higher as compared to other hepatocytes.     

Attributes of atmospheric carbon monoxide oxidation by Maine forest soils

CO, one of the most important trace gases, regulates tropospheric methane, hydroxyl radical, and ozone contents. Ten to 25% of the estimated global CO flux may be consumed by soils annually. Depth profiles for (14)CO oxidation and CO concentration indicated that CO oxidation occurred primarily in surface soils and that photooxidation of soil organic matter did not necessarily contribute significantly to CO fluxes. Kinetic analyses revealed that the apparent K(m) was about 18 nM (17 ppm) and the V(max) was 6.9 micromol g (fresh weight)(-1) h(-1); the apparent K(m) was similar to the apparent K(m) for atmospheric methane consumption, but the V(max) was more than 100 times higher. Atmospheric CO oxidation responded sensitively to soil water regimes; decreases in water content in initially saturated soils resulted in increased uptake, and optimum uptake occurred at water contents of 30 to 60%. However, extended drying led to decreased uptake and net CO production. Rewetting could restore CO uptake, albeit with a pronounced hysteresis. The responses to changing temperatures indicated that the optimum temperature for net uptake was between 20 and 25 degrees C and that there was a transition to net production at temperatures above 30 degrees C. The responses to methyl fluoride and acetylene indicated that populations other than ammonia oxidizers and methanotrophs must be involved in forest soils. The response to acetylene was notable, since the strong initial inhibition was reversed after 12 h of incubation; in contrast, methyl fluoride did not have an inhibitory effect. Ammonium did not inhibit CO uptake; the level of nitrite inhibition was initially substantial, but nitrite inhibition was reversible over time. Nitrite inhibition appeared to occur through indirect effects based on abiological formation of NO.     

Initial net CO2 uptake responses and root growth for a CAM community placed in a closed environment

To help understand carbon balance between shoots and developing roots, 41 bare-root crassulacean acid metabolism (CAM) plants native to the Sonoran Desert were studied in a glass-panelled sealable room at day/night air temperatures of 25/15 degrees C. Net CO(2) uptake by the community of Agave schottii, Carnegia gigantea, Cylindropuntia versicolor, Ferocactus wislizenii and Opuntia engelmannii occurred 3 weeks after watering. At 4 weeks, the net CO(2) uptake rate measured for south-east-facing younger parts of the shoots averaged 1.94 micro mol m(-2) s(-1) at night, considerably higher than the community-level nocturnal net CO(2) uptake averaged over the total shoot surface, primarily reflecting the influences of surface orientation on radiation interception (predicted net CO(2) uptake is twice as high for south-east-facing surfaces compared with all compass directions). Estimated growth plus maintenance respiration of the roots averaged 0.10 micro mol m(-2) s(-1) over the 13-week period, when the community had a net carbon gain from the atmosphere of 4 mol C while the structural C incorporated into the roots was 23 mol. Thus, these five CAM species diverted all net C uptake over the 13-week period plus some existing shoot C to newly developing roots. Only after sufficient roots develop to support shoot water and nutrient requirements will the plant community have net above-ground biomass gains.     

Temperature as a control over ecosystem CO<Subscript>2</Subscript> fluxes in a high-elevation,subalpine forest

We evaluated the hypothesis that CO(2) uptake by a subalpine, coniferous forest is limited by cool temperature during the growing season. Using the eddy covariance approach we conducted observations of net ecosystem CO(2) exchange (NEE) across two growing seasons. When pooled for the entire growing season during both years, light-saturated net ecosystem CO(2) exchange (NEE(sat)) exhibited a temperature optimum within the range 7-12 degrees C. Ecosystem respiration rate ( R(e)), calculated as the y-intercept of the NEE versus photosynthetic photon flux density (PPFD) relationship, increased with increasing temperature, causing a 15% reduction in net CO(2) uptake capacity for this ecosystem as temperatures increased from typical early season temperatures of 7 degrees C to typical mid-season temperatures of 18 degrees C. The ecosystem quantum yield and the ecosystem PPFD compensation point, which are measures of light-utilization efficiency, were highest during the cool temperatures of the early season, and decreased later in the season at higher temperatures. Branch-level measurements revealed that net photosynthesis in all three of the dominant conifer tree species exhibited a temperature optimum near 10 degrees C early in the season and 15 degrees C later in the season. Using path analysis, we statistically isolated temperature as a seasonal variable, and identified the dynamic role that temperature exhibits in controlling ecosystem fluxes early and late in the season. During the spring, an increase in temperature has a positive effect on NEE, because daytime temperatures progress from near freezing to near the photosynthetic temperature optimum, and R(e )values remain low. During the middle of the summer an increase in temperature has a negative effect on NEE, because inhibition of net photosynthesis and increases in R(e). When taken together, the results demonstrate that in this high-elevation forest ecosystem CO(2) uptake is not limited by cool-temperature constraints on photosynthetic processes during the growing-season, as suggested by some previous ecophysiological studies at the branch and needle levels. Rather, it is warm temperatures in the mid-summer, and their effect on ecosystem respiration, that cause the greatest reduction in the potential for forest carbon sequestration.     

Effects of alpha-Hydroxy-2-Pyridinemethanesulfonic Acid on Photosynthetic Carbon Dioxide Uptake and Stomatal Movements in Excised Tomato Leaves

The effects of 10(-2)m alpha-hydroxy-2-pyridinemethanesulfonic acid (alphaHPMS) on the CO(2) compensation point, photosynthetic CO(2) uptake, CO(2) evolution into CO(2)-free air in light, and stomatal movement, in excised tomato leaves (Lycopersicon esculentum Mill. Eurocross BB-F(1) Hybrid) were studied. It was found that alpha-HPMS had a transient lowering effect on the CO(2) compensation point of treated leaves within the first 5 minutes of application. The net photosynthetic CO(2) uptake was inhibited by alpha-HPMS treatment. The inhibition increased with time and was enhanced in an O(2)-free atmosphere. The CO(2) evolution into CO(2)-free air in light was inhibited by alpha-HPMS. The inhibition was O(2)-dependent because the effect was observed only in 21% O(2) but not in O(2)-free N(2). Stomatal apertures were affected by alpha-HPMS, but the effect was transient and was observed 15 to 30 minutes after the application. The time course of this closure did not account for the observed inhibition of net CO(2) uptake.     

The effect of induced heat waves on Pinus taeda and Quercus rubra seedlings in ambient and elevated CO(2) atmospheres

Here, we investigated the effect of different heat-wave intensities applied at two atmospheric CO(2) concentrations ([CO(2) ]) on seedlings of two tree species, loblolly pine (Pinus taeda) and northern red oak (Quercus rubra). Seedlings were assigned to treatment combinations of two levels of [CO(2) ] (380 or 700?μmol?mol(-1) ) and four levels of air temperature (ambient, ambient +3°C, or 7-d heat waves consisting of a biweekly +6°C heat wave, or a monthly +12°C heat wave). Treatments were maintained throughout the growing season, thus receiving equal heat sums. We measured gas exchange and fluorescence parameters before, during and after a mid-summer heat wave. The +12°C heat wave, significantly reduced net photosynthesis (A(net) ) in both species and [CO(2) ] treatments but this effect was diminished in elevated [CO(2) ]. The decrease in A(net) was accompanied by a decrease in F(v) '/F(m) ' in P.?taeda and Φ(PSII) in Q.?rubra. Our findings suggest that, if soil moisture is adequate, trees will experience negative effects in photosynthetic performance only with the occurrence of extreme heat waves. As elevated [CO(2) ] diminished these negative effects, the future climate may not be as detrimental to plant communities as previously assumed.     

Enhancement of rice canopy carbon gain by elevated CO(2) is sensitive to growth stage and leaf nitrogen concentration

Increasing our understanding of the factors regulating seasonal changes in rice canopy carbon gain (C(gain): daily net photosynthesis -- night respiration) under elevated CO(2) concentrations ([CO(2)]) will reduce our uncertainty in predicting future rice yields and assist in the development of adaptation strategies. In this study we measured CO(2) exchange from rice (Oryza sativa) canopies grown at c. 360 and 690 micromol mol(-1)[CO(2)] in growth chambers continuously over three growing seasons. Stimulation of C(gain) by elevated [CO(2)] was 22-79% during vegetative growth, but decreased to between -12 and 5% after the grain-filling stage, resulting in a 7-22% net enhancement for the whole season. The decreased stimulation of C(gain) resulted mainly from decreased canopy net photosynthesis and partially from increased respiration. A decrease in canopy photosynthetic capacity was noted where leaf nitrogen (N) decreased. The effect of elevated [CO(2)] on leaf area was generally small, but most dramatic under ample N conditions; this increased the stimulation of whole-season C(gain). These results suggest that a decrease in C(gain) enhancement following elevated CO(2) levels is difficult to avoid, but that careful management of nitrogen levels can alter the whole-season C(gain) enhancement.     

Limited Predictability of Amino Acid Substitutions in Seasonal Influenza Viruses

Seasonal influenza viruses repeatedly infect humans in part because they rapidly change their antigenic properties and evade host immune responses, necessitating frequent updates of the vaccine composition. Accurate predictions of strains circulating in the future could therefore improve the vaccine match. Here, we studied the predictability of frequency dynamics and fixation of amino acid substitutions. Current frequency was the strongest predictor of eventual fixation, as expected in neutral evolution. Other properties, such as occurrence in previously characterized epitopes or high Local Branching Index (LBI) had little predictive power. Parallel evolution was found to be moderately predictive of fixation. Although the LBI had little power to predict frequency dynamics, it was still successful at picking strains representative of future populations. The latter is due to a tendency of the LBI to be high for consensus-like sequences that are closer to the future than the average sequence. Simulations of models of adapting populations, in contrast, show clear signals of predictability. This indicates that the evolution of influenza HA and NA, while driven by strong selection pressure to change, is poorly described by common models of directional selection such as traveling fitness waves.     

Environment or development? Lifetime net CO2 exchange and control of the expression of Crassulacean acid metabolism in Mesembryanthemum crystallinum

The relative influence of plant age and environmental stress signals in triggering a shift from C(3) photosynthesis to Crassulacean acid metabolism (CAM) in the annual halophytic C(3)-CAM species Mesembryanthemum crystallinum was explored by continuously monitoring net CO(2) exchange of whole shoots from the seedling stage until seed set. Plants exposed to high salinity (400 mm NaCl) in hydroponic culture solution or grown in saline-droughted soil acquired between 11% and 24% of their carbon via net dark CO(2) uptake involving CAM. In contrast, plants grown under nonsaline, well-watered conditions were capable of completing their life cycle by operating in the C(3) mode without ever exhibiting net CO(2) uptake at night. These observations are not consistent with the widely expressed view that the induction of CAM by high salinity in M. crystallinum represents an acceleration of preprogrammed developmental processes. Rather, our study demonstrates that the induction of the CAM pathway for carbon acquisition in M. crystallinum is under environmental control.     

Impairment of C(4) photosynthesis by drought is exacerbated by limiting nitrogen and ameliorated by elevated [CO(2)] in maize

Predictions of future ecosystem function and food supply from staple C(4) crops, such as maize, depend on elucidation of the mechanisms by which environmental change and growing conditions interact to determine future plant performance. To test the interactive effects of elevated [CO(2)], drought, and nitrogen (N) supply on net photosynthetic CO(2) uptake (A) in the world's most important C(4) crop, maize (Zea mays) was grown at ambient [CO(2)] (～385 ppm) and elevated [CO(2)] (550 ppm) with either high N supply (168 kg N ha(-1) fertilizer) or limiting N (no fertilizer) at a site in the US Corn Belt. A mid-season drought was not sufficiently severe to reduce yields, but caused significant physiological stress, with reductions in stomatal conductance (up to 57%), A (up to 44%), and the in vivo capacity of phosphoenolpyruvate carboxylase (up to 58%). There was no stimulation of A by elevated [CO(2)] when water availability was high, irrespective of N availability. Elevated [CO(2)] delayed and relieved both stomatal and non-stomatal limitations to A during the drought. Limiting N supply exacerbated stomatal and non-stomatal limitation to A during drought. However, the effects of limiting N and elevated [CO(2)] were additive, so amelioration of stress by elevated [CO(2)] did not differ in magnitude between high N and limiting N supply. These findings provide new understanding of the limitations to C(4) photosynthesis that will occur under future field conditions of the primary region of maize production in the world.     

Evidence for an inorganic carbon-concentrating mechanism in the symbiotic dinoflagellate Symbiodinium sp

The presence of a carbon-concentrating mechanism in the symbiotic dinoflagellate Symbiodinium sp. was investigated. Its existence was postulated to explain how these algae fix inorganic carbon (C(i)) efficiently despite the presence of a form II Rubisco. When the dinoflagellates were isolated from their host, the giant clam (Tridacna gigas), CO(2) uptake was found to support the majority of net photosynthesis (45%-80%) at pH 8.0; however, 2 d after isolation this decreased to 5% to 65%, with HCO(3)(-) uptake supporting 35% to 95% of net photosynthesis. Measurements of intracellular C(i) concentrations showed that levels inside the cell were between two and seven times what would be expected from passive diffusion of C(i) into the cell. Symbiodinium also exhibits a distinct light-activated intracellular carbonic anhydrase activity. This, coupled with elevated intracellular C(i) and the ability to utilize both CO(2) and HCO(3)(-) from the medium, suggests that Symbiodinium sp. does possess a carbon-concentrating mechanism. However, intracellular C(i) levels are not as large as might be expected of an alga utilizing a form II Rubisco with a poor affinity for CO(2).     

Isolation and characterization of mutants of common ice plant deficient in crassulacean acid metabolism

Crassulacean acid metabolism (CAM) is a specialized mode of photosynthesis that improves water use efficiency by shifting part or all of net atmospheric CO2 uptake to the night. Genetic dissection of regulatory and metabolic attributes of CAM has been limited by the difficulty of identifying a reliable phenotype for mutant screening. We developed a novel and simple colorimetric assay to measure leaf pH to screen fast neutron-mutagenized populations of common ice plant (Mesembryanthemum crystallinum), a facultative CAM species, to detect CAM-deficient mutants with limited nocturnal acidification. The isolated CAM-deficient mutants showed negligible net dark CO2 uptake compared with wild-type plants following the imposition of salinity stress. The mutants and wild-type plants accumulated nearly comparable levels of sodium in leaves, but the mutants grew more slowly than the wild-type plants. The mutants also had substantially reduced seed set and seed weight relative to wild type under salinity stress. Carbon-isotope ratios of seed collected from 4-month-old plants indicated that C3 photosynthesis made a greater contribution to seed production in mutants compared to wild type. The CAM-deficient mutants were deficient in leaf starch and lacked plastidic phosphoglucomutase, an enzyme critical for gluconeogenesis and starch formation, resulting in substrate limitation of nocturnal C4 acid formation. The restoration of nocturnal acidification by feeding detached leaves of salt-stressed mutants with glucose or sucrose supported this defect and served to illustrate the flexibility of CAM. The CAM-deficient mutants described here constitute important models for exploring regulatory features and metabolic consequences of CAM.     

Through enhanced tree dynamics carbon dioxide enrichment may cause tropical forests to lose carbon

The fixation and storage of C by tropical forests, which contain close to half of the globe's biomass C, may be affected by elevated atmospheric CO2 concentration. Classical theoretical approaches assume a uniform stimulation of photosynthesis and growth across taxa. Direct assessments of the C balance either by flux studies or by repeated forest inventories also suggest a current net uptake, although magnitudes sometimes exceed those missing required to balance the global C cycle. Reasons for such discrepancies may lie in the nature of forest dynamics and in differential responses of taxa or plant functional types. In this contribution I argue that CO2 enrichment may cause forests to become more dynamic and that faster tree turnover may in fact convert a stimulatory effect of elevated CO2 on photosynthesis and growth into a long-term net biomass C loss by favouring shorter-lived trees of lower wood density. At the least, this is a scenario that deserves inclusion into long-term projections of the C relations of tropical forests. Species and plant functional type specific responses ('biodiversity effects') and forest dynamics need to be accounted for in projections of future C storage and cycling in tropical forests.     

Crassulacean acid metabolism in the ZZ plant, Zamioculcas zamiifolia (Araceae)

Zamioculcas zamiifolia (Araceae), a terrestrial East African aroid, with two defining attributes of crassulacean acid metabolism (CAM) (net CO(2) uptake in the dark and diel fluctuations of titratable acidity) is the only CAM plant described within the Araceae, a mainly tropical taxon that contains the second largest number of epiphytes of any vascular plant family. Within the Alismatales, the order to which the Araceae belong, Z. zamiifolia is the only documented nonaquatic CAM species. Zamioculcas zamiifolia has weak CAM that is upregulated in response to water stress. In well-watered plants, day-night fluctuations in titratable acidity were 2.5 μmol H(+)·(g fresh mass)(-1), and net CO(2) uptake in the dark contributed less than 1% to daily carbon gain. Following 10 d of water stress, net CO(2) uptake in the light fell 94% and net CO(2) uptake in the dark increased 7.5-fold, such that its contribution increased to 19% of daily carbon gain. Following rewatering, dark CO(2) uptake returned to within 5% of prestressed levels. We postulate that CAM assists survival of Z. zamiifolia by reducing water loss and maintaining carbon gain during seasonal droughts characteristic of its natural habitat.     

Evolutionary context for understanding and manipulating plant responses to past, present and future atmospheric [CO2

Variation in atmospheric [CO(2)] is a prominent feature of the environmental history over which vascular plants have evolved. Periods of falling and low [CO(2)] in the palaeo-record appear to have created selective pressure for important adaptations in modern plants. Today, rising [CO(2)] is a key component of anthropogenic global environmental change that will impact plants and the ecosystem goods and services they deliver. Currently, there is limited evidence that natural plant populations have evolved in response to contemporary increases in [CO(2)] in ways that increase plant productivity or fitness, and no evidence for incidental breeding of crop varieties to achieve greater yield enhancement from rising [CO(2)]. Evolutionary responses to elevated [CO(2)] have been studied by applying selection in controlled environments, quantitative genetics and trait-based approaches. Findings to date suggest that adaptive changes in plant traits in response to future [CO(2)] will not be consistently observed across species or environments and will not be large in magnitude compared with physiological and ecological responses to future [CO(2)]. This lack of evidence for strong evolutionary effects of elevated [CO(2)] is surprising, given the large effects of elevated [CO(2)] on plant phenotypes. New studies under more stressful, complex environmental conditions associated with climate change may revise this view. Efforts are underway to engineer plants to: (i) overcome the limitations to photosynthesis from today's [CO(2)] and (ii) benefit maximally from future, greater [CO(2)]. Targets range in scale from manipulating the function of a single enzyme (e.g. Rubisco) to adding metabolic pathways from bacteria as well as engineering the structural and functional components necessary for C(4) photosynthesis into C(3) leaves. Successfully improving plant performance will depend on combining the knowledge of the evolutionary context, cellular basis and physiological integration of plant responses to varying [CO(2)].     

Functional hepatocyte heterogeneity. Vascular 2-oxoglutarate is almost exclusively taken up by perivenous, glutamine-synthetase-containing hepatocytes

1. In isolated perfused rat liver maximal rates of 2-[1-14C]oxoglutarate uptake were about 0.4 mumol.g-1 .min-1; half-maximal rates of 2-[14C]oxoglutarate uptake were observed with influent concentrations of about 100 microM. 2-[14C]Oxoglutarate uptake by the liver was not affected by the direction of perfusion, but was decreased by about 80-90% when Na+ in the perfusion fluid was substituted by choline+, suggesting a Na+-dependence of hepatic 2-oxoglutarate uptake. In the absence of added ammonia, [14C]oxoglutarate uptake by the liver was about twice the net oxoglutarate uptake, indicating a simultaneous release of unlabeled oxoglutarate from perfused rat liver. 2. 14C-Labeled metabolites derived from [1-14C]oxoglutarate and recovered in the effluent perfusate were 14CO2 and 14C-labeled glutamate and glutamine; they accounted for 85-100% of the radiolabel taken up by the liver. 14CO2 was the major product (more than 70%) from [1-14C]oxoglutarate taken up the liver, provided glutamine synthesis was either inhibited by methionine sulfoximine or the endogenous rate of glutamine production was below 40 nmol.g-1.min-1. 3. Stimulation of glutamine synthesis by ammonia did not affect [14C]oxoglutarate uptake by the liver, but considerably increased net hepatic oxoglutarate uptake, indicating a decreased release of unlabeled oxoglutarate from the liver. Stepwise stimulation of hepatic glutamine synthesis led to a gradual decrease of 14CO2 production and radiolabel was recovered increasingly as [14C]glutamine in the effluent. At high rates of glutamine formation (i.e. about 0.6 mumol.g-1.min-1), about 60% of the [1-14C]oxoglutarate taken up by the liver was recovered in the effluent as [14C]glutamine. 14CO2 and [14C]glutamine production from added [1-14C]oxoglutarate were dependent on the rate of hepatic glutamine synthesis but not on the direction of perfusion. Extrapolation of 14C incorporation into glutamine to maximal rates of hepatic glutamine synthesis yielded an about 100% utilization of the [14C]oxoglutarate taken up by the liver for glutamine synthesis. This was again true for both the antegrade and the retrograde perfusion directions. On the other hand, addition of ammonia did not affect 14CO2 production from labeled oxoglutarate, when glutamine synthetase was inhibited by methionine sulfoximine. 4. The data suggest that vascular oxoglutarate is almost exclusively taken up by the small perivenous hepatocyte population containing glutamine synthetase, i.e. a cell population comprising only 6-7% of all hepatocytes. Thus, the findings demonstrate the existence of a, to date, uniquely zonally distributed oxoglutarate transport system which is probably Na+-dependent in the plasma membrane.(ABSTRACT TRUNCATED AT 400 WORDS)