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Energy and water requirements of lactation in the North American porcupine, Erethizon dorsatum.

1. Energy and water requirements of lactating porcupines were compared with results of previous studies on energetics of reproduction in small-bodied rodents. 2. Mass-specific food and water intake of control and lactating porcupines was examined throughout the 68-78 days of lactation. Water intake of lactating females was 16% higher than that of non-lactating animals. 3. Digestive efficiency of porcupines fed commercial rabbit chow was 54-60%; there was no significant difference in efficiency between lactating and non-lactating animals. 4. Total mean energetic intake throughout lactation was only 17% greater than that of non-lactating animals. 5. Reproductive rate and costs of lactation in porcupines are considerably less than in other rodents and other comparably-sized mammals, but the amount of energy allocated to each offspring is quite high. 6. The reproductive pattern of porcupines is associated with low juvenile mortality and long adult lifespan (both of which reflect the porcupine's protective morphology), and may be related to the quality of winter diets.

Animals↗

Developmental regulation of Sertoli cell lactate production by hormones and the testicular paracrine factor, PModS.

Testicular peritubular cells produce a paracrine factor termed PModS that mediates mesenchymal-epithelial interactions and modulates Sertoli cell functions essential for the process of spermatogenesis. Sertoli cells produce lactate as a preferred energy metabolite for developing spermatogenic cells. The current study was designed to examine the actions of PModS and hormones on Sertoli cell lactate production at various stages of pubertal development. Sertoli cells were isolated from pre-pubertal (10 day), mid-pubertal (20 day) and late pubertal (35 day) rat testes. Lactate accumulation in the conditioned-medium of cultured Sertoli cells was measured. Basal lactate production increased approximately fivefold during pubertal Sertoli cell development. Therefore, lactate production increases as the Sertoli cell differentiates during pubertal development. The ability of regulatory agents such as FSH or a combination of FSH, insulin, retinol and testosterone (FIRT) to stimulate lactate production decreased during pubertal development as Sertoli cell differentiation increased. Purified PModS stimulated lactate production in Sertoli cell preparations throughout pubertal development. PModS had a greater effect than FSH in stimulating late pubertal Sertoli cell lactate production. PModS in combination with FIRT resulted in an additive stimulation of lactate production suggesting a distinct mechanism of action for PModS. Observations support the proposal that the locally produced paracrine factor PModS mediates mesenchymal-epithelial cell interactions during pubertal development and that these interactions promote Sertoli cell differentiated functions (i.e. lactate production) required for the developing spermatogenic cells.

Animals↗

Transport and metabolism of D-lactate in Jerusalem artichoke mitochondria.

We report here initial studies on D-lactate metabolism in Jerusalem artichoke. It was found that: 1) D-lactate can be synthesized by Jerusalem artichoke by virtue of the presence of glyoxalase II, the activity of which was measured photometrically in both isolated Jerusalem artichoke mitochondria and cytosolic fraction after the addition of S-D-lactoyl-glutathione. 2) Externally added D-lactate caused oxygen consumption by mitochondria, mitochondrial membrane potential increase and proton release, in processes that were insensitive to rotenone, but inhibited by both antimycin A and cyanide. 3) D-lactate was metabolized inside mitochondria by a flavoprotein, a putative D-lactate dehydrogenase, the activity of which could be measured photometrically in mitochondria treated with Triton X-100. 4) Jerusalem artichoke mitochondria can take up externally added D-lactate by means of a D-lactate/H(+) symporter investigated by measuring the rate of reduction of endogenous flavins. The action of the d-lactate translocator and of the mitochondrial D-lactate dehydrogenase could be responsible for the subsequent metabolism of d-lactate formed from methylglyoxal in the cytosol of Jerusalem artichoke.

Biological Transport↗

Detection of intracellular lactate with localized diffusion {1H-13C}-spectroscopy in rat glioma in vivo.

The aim of this study was to compare the diffusion characteristic of lactate and alanine in a brain tumor model to that of normal brain metabolites known to be mainly intracellular such as N-acetylaspartate or creatine. The diffusion of (13)C-labeled metabolites was measured in vivo with localized NMR spectroscopy at 9.4 T (400 MHz) using a previously described localization and editing pulse sequence known as ACED-STEAM ('adiabatic carbon editing and decoupling'). (13)C-labeled glucose was administered and the apparent diffusion coefficients of the glycolytic products, {(1)H-(13)C}-lactate and {(1)H-(13)C}-alanine, were determined in rat intracerebral 9L glioma. To obtain insights into {(1)H-(13)C}-lactate compartmentation (intra- versus extracellular), the pulse sequence used very large diffusion weighting (50 ms/microm(2)). Multi-exponential diffusion attenuation of the lactate metabolite signals was observed. The persistence of a lactate signal at very large diffusion weighting provided direct experimental evidence of significant intracellular lactate concentration. To investigate the spatial distribution of lactate and other metabolites, (1)H spectroscopic images were also acquired. Lactate and choline-containing compounds were consistently elevated in tumor tissue, but not in necrotic regions and surrounding normal-appearing brain. Overall, these findings suggest that lactate is mainly associated with tumor tissue and that within the time-frame of these experiments at least some of the glycolytic product ([(13)C] lactate) originates from an intracellular compartment.

Alanine↗

Serial blood lactate levels can predict the development of multiple organ failure following septic shock.

BACKGROUND: Despite successful initial resuscitation, septic shock frequently evolves into multiple system organ failure (MSOF) and death. Since blood lactate levels can reflect the degree of cellular derangements, we examined the relation between serial blood lactate levels and the development of MSOF, or mortality, in patients with septic shock. PATIENTS AND METHODS: In 87 patients with a first episode of septic shock, we measured initial lactate (at onset of septic shock), final lactate (before recovery or death), "lactime" (time during which blood lactate was > 2.0 mmol/L, and the area under the curve (AUC) for abnormal values (above 2.0 mmol/L). These measurements were correlated with survival and organ failure and scored for four systems (ie, respiratory, renal, hepatic, and coagulation), adding to a maximal score of 8. RESULTS: Thirty-three (38%) patients survived. Of the 54 (62%) nonsurvivors, the 13 patients who died during the first 24 hours of septic shock had higher initial blood lactate levels than those who died later (mean +/- standard deviation 9.6 +/- 5.3 mmol/L versus 5.6 +/- 3.7 mmol/L, P< 0.05). The 74 patients who survived the first 24 hours of shock, were studied in more detail. On presentation, survivors had a significantly higher mean arterial pressure (76 +/- 12 mm Hg versus 63 +/- 20 mm Hg, P < 0.001) and arterial pH (7.40 +/- 0.07 versus 7.37 +/- 0.09, P< 0.05) than nonsurvivors. Although the differences in initial blood lactate levels between survivors and nonsurvivors did not reach statistical significance (4.7 +/- 2.5 mmol/L versus 5.6 +/- 3.7 mmol/L), only the survivors had a significant decrease during the first 24 hours of septic shock. The survivors had a significantly lower lactime and AUC than the nonsurvivors. The duration of lactic acidosis was the best predictor of survival (multiple regression analysis, R2 = 0.266, P <0.001), followed by age, heart rate, and mean arterial pressure. Patients with lower organ failure scores had lower initial blood lactate, lactime, and AUC. The duration of lactic acidosis was the only significant predictor of organ failure. CONCLUSIONS: In patients with septic shock, serial determinations of blood lactate levels are good predictors of the development of MSOF an death. In this respect, the duration of lactic acidosis is more important than the initial lactate value. Although a number of factors may contribute to hyperlactatemia, these observations are compatible with a direct role of prolonged tissue hypoxia in the development of complications following septic shock.

Adult↗

Metabolic mapping of the brain in pregnant, parturient and lactating rats using fos immunohistochemistry.

The present study was designed to investigate Fos-positive neurons of the female rat brain at various reproductive states in order to analyze the metabolic map connected with pregnancy, parturition and lactation. The number of Fos-positive neurons in each brain nucleus was analyzed with a quantitative immunohistochemical method in virgin, pregnant, parturient, lactating and arrested lactating rats. In parturient rats, a significant number of Fos-positive neurons was observed as compared to virgin or pregnant females in the following brain regions; the bed nucleus of the stria terminalis (BST), lateral septal nucleus (LS), medial preoptic area (MPA), periventricular hypothalamic nucleus (Pe), parvocellular paraventricular hypothalamic nucleus (PaPVN), magnocellular paraventricular hypothalamic nucleus (MaPVN), supraoptic nucleus (SON), paraventricular thalamic nucleus (PV), anterior hypothalamic area (AHA), lateral hypothalamic area (LH), amygdaloid nucleus (AM), supramammillary nucleus (SuM), substantia nigra (SN), central grey (CG), microcellular tegmental nucleus (MiTg), subparafascicular thalamic nucleus (SPF), posterior hypothalamic area (PH), dorsal raphe nucleus (DR), locus coeruleus (LC), dorsal parabrachial nucleus (DPB), nucleus of solitary tract (Sol), and ventrolateral medulla (VLM). Significant differences were found in the number of Fos-positive neurons between parturient and lactating females, although localization of Fos-positive neurons in lactating females was quite similar to parturient ones. Between parturient and lactating rats: (1) In the MPA, PaPVN, AHA, arcuate hypothalamic nucleus (Arc), ventromedial hypothalamic nucleus (VMH), MLT, and Ge, the number of Fos-positive neurons of lactating females were significantly higher than those of parturient ones; (2) In the LS, Pe, PV, LH, AM, SuM, CG, MiTg, SPF, PH, DR, LC, and VLM, there was no significant differences in the number of Fos-positive neurons; (3) In the BST, MaPVN, SON, SN, DPB and Sol, the number of Fos-positive neurons of lactating rats were significantly lower than those of parturient ones. These different patterns of Fos expression among many brain regions may be owing to the functional differences in each region. Fos expression in lactating rats was apparently induced by suckling stimulation because the removal of their litters immediately after parturition completely eliminated expression of Fos protein in each nucleus. These results suggest that the localization of Fos-positive neurons in a number of neural populations throughout the brain may be revealing the neural circuits in response to parturition or lactation.

Animals↗

The value of blood lactate measurements in ICU: an evaluation of the role in the management of patients on haemofiltration.

In response to clinical demand some point-of-care analysers now provide blood lactate measurements, but recently concern has been expressed about the value and interpretation of these measurements. We undertook this study to evaluate blood lactate measurements in patients with acute renal failure undergoing haemofiltration (HF) with lactate replacement fluid. At baseline, 27 patients had base deficits of >5 mmol/l and 14 (52%) had blood lactates of >3.5 mmol/l. Lactate 'tolerance' was monitored by peak changes in these parameters during the procedure. There was a worsening of base deficit in only three of the patients in whom lactate rises exceeded 10 mmol/l with one survivor. Twelve patients with rises of blood lactate greater than 5 mmol/l improved their base deficit (+1 to +17) with eight (67%) survivors. Of the remaining 12 patients with improved base deficit (+2 to +20), 10 (83%) survived. Lactate tolerance was compromised in patients with co-incidental liver disease, those on inotropic support, and in patients with initial blood lactate measurements of >10 mmol/l and large base deficits. The data suggest that blood lactate and simultaneous acid-base response measurements during HF help to assign correct buffer replacement and should be performed on all patients.

Acute Kidney Injury↗

Adenomas of the breast and ectopic breast under lactational influences.

The tubular adenomas, lactating adenomas, and ectopic lactation- or gestation-associated benign lesions of the breast diagnosed during a 22-year period are reported. Forty-two breast adenomas (in 28 patients) that demonstrated lactational changes are reviewed, with special attention to classification and clinical presentation. Five ectopic lactating adenomas--three axillary or in the chest wall and two vulvar, the latter occurring in the same patient--were identified in three patients. The lesions in the remaining cases were fibroadenomas with lactational changes (16 patients) and lactating or tubular adenomas (nine patients). Striking histologic and clinical similarities between the tubular and lactating adenomas and the lesions identified as lactational changes within a fibroadenoma were observed, with foci of tubular adenomatous change present within otherwise typical fibroadenomas. The overlapping morphologic spectrum and the strong association with pregnancy or lactational influences blur the distinction between these entities and suggest a common pathogenesis. The cytologic features of lactational lesions of the breast and ectopic breast pose special problems in diagnosis by fine needle aspiration.

Adenofibroma↗

Lactic acid and protein interactions: implications for the NMR visibility of lactate in biological systems.

The addition of bovine serum albumin (BSA) to a solution of lactate and alanine resulted in the disappearance of the 1H-NMR resonances from lactate but not alanine. As temperature is increased lactate becomes increasingly NMR visible and after heating above 65 degreesC and cooling to 25 degreesC lactate binding is reduced. With a concentration of 0.2 mM BSA, there was a linear relationship between NMR visible lactate versus total lactate over a range of lactate concentrations of 0.2-35 mM (slope 0.384+/-0.003) indicating that approx. 60% of the added lactate is not visible in the 1H-NMR spectrum. With a 0.1 mM BSA solution, however, the slope was markedly higher indicating that under these conditions only 25-30% of the lactate was NMR invisible. The results from this study indicate that decreased NMR visibility of lactate in proteinaceous solutions is due to non-specific binding which is dependent on the tertiary structure of the protein. This has important implications not only for the interpretation of in vivo 1H-NMR experiments but also for 13C, and 14C studies of metabolism.

Lactates↗

Differential messenger RNA distribution of lactate dehydrogenase LDH-1 and LDH-5 isoforms in the rat brain.

The role of lactate in brain energy metabolism has recently received renewed attention. Although blood-borne monocarboxylates such as lactate poorly cross the blood-brain barrier in the adult brain, lactate produced within the brain parenchyma may be a suitable substrate for brain cells. Lactate dehydrogenase is crucial for both the production and utilization of lactate. In this article, we report the regional distribution of the messenger RNAs for lactate dehydrogenase isoforms 1 and 5 in the adult rat brain using in situ hybridization histochemistry with specific [alpha-(35)S]dATP 3' end-labeled oligoprobes. The autoradiographs revealed that the lactate dehydrogenase-1 messenger RNA is highly expressed in a variety of brain structures, including the main olfactory bulb, the piriform cortex, several thalamic and hypothalamic nuclei, the pontine nuclei, the ventral cochlear nucleus, the trigeminal nerve and the solitary tractus nucleus. In addition, the granular and Purkinje cell layers of the cerebellum showed a strong labeling. The neocortex (e.g., cingular, retrosplenial and frontoparietal cortices) often exhibits a marked laminar pattern of distribution of lactate dehydrogenase-1 messenger RNA (layers II/III, IV and VI being most strongly labeled). In contrast, expression of the lactate dehydrogenase-5 messenger RNA generally seemed more diffusely distributed across the different brain regions. Expression was particularly strong in the hippocampal formation (especially in Ammon's horn and dentate gyrus) and in the cerebral cortex, where no laminar pattern of distribution was observed. Overall, these data are consistent with the emerging idea that lactate is an important energy substrate produced and consumed by brain cells.

Animals↗

Effects of lactation on metabolic and reproductive hormones in Lipizzaner mares.

In this study, growth hormone (GH), insulin-like growth factor 1 (IGF-1), leptin, luteinising hormone (LH) and prolactin were analyzed in mares from late pregnancy throughout lactation (group 1, n=46) and in non-lactating mares (group 2, n=11). Plasma GH concentrations in group 1 mares during gestation and lactation were lower than in mares of group 2 (P<0.05). Highest IGF-1 levels were found in lactating mares in the week of foaling. IGF-1 concentrations decreased continuously thereafter. Plasma leptin concentrations decreased after foaling and, for 4 weeks, were lower in lactating than in non-lactating mares (P<0.05). Reduced leptin concentrations may promote feed intake and allow lactating mares to avoid an energy deficit. In group 1 mares, prolactin concentrations reached a maximum in the week of foaling and decreased rapidly thereafter. Plasma LH concentrations in group 1 mares before foaling were lower than at corresponding times in group 2 (P<0.05). LH concentrations then increased and did no longer differ from group 2 until week 2 postpartum. This increase may contribute to the resumption of cyclic ovarian activity in postpartum mares. Subsequently, LH levels in lactating mares decreased again (P<0.05). Increased IGF-1 concentrations early postpartum might contribute to ovarian stimulation while reduced IGF-1 and GH concentrations later in lactation might cause reduced stimulation. The changes in somatotrophic hormones could thus explain, at least in part, a more pronounced stimulation of ovarian function early postpartum than during the following months of lactation.

Animals↗

Respiratory burst activity of blood and milk neutrophils in dairy cows during different stages of lactation.

The non-stimulated and phorbol 12-myristate 13-acetate (PMA)-stimulated luminol-augmented cellular chemiluminescence (CL) response and viability of milk and blood polymorphonuclear leukocytes (PMN) were determined in lactating dairy cows during different stages of lactation. In the first study, ten healthy cows each in early, mid and late lactation were compared. In a second study, the same measurements as in the first study were evaluated longitudinally in 12 cows during 1 month following parturition. The CL activity and myeloperoxidase (MPO) content of milk PMN and macrophages (M) were also compared. Milk M did not possess MPO activity and were devoid of any luminol-enhanced CL. The CL activity of milk and blood PMN was significantly lower in early lactation than in mid and late lactation (P < 0.001). Whereas little changes were observed in viability of blood PMN, the viability of milk PMN was lower in early lactation than in mid and late lactation (P < 0.001). The percentage of PMN in isolated milk cells was also lower during early lactation than during mid and late lactation (P < 0.001). The CL activity in response to PMA during early, mid and late lactation increased 13, 59 and 42-fold in blood PMN and 1.7, 2.6 and 2.4-fold in milk PMN, respectively, in comparison with non-stimulated PMN. The CL activity, both in milk and blood PMN. the milk PMN viability and the percentage of milk PMN were lowest between 3 d and 11 d post partum. These observed changes immediately after calving could contribute to a higher susceptibility to mastitis in that period.

Animals↗

Effects of stage of lactation, milk protein genotype and body condition at calving on protein composition and renneting properties of bovine milk.

The lactational variation in milk protein composition and renneting properties and their relationship to the cow's body condition at calving were investigated in 39 Danish Holstein first lactation cows fed on a well balanced standard diet. All milk characteristics measured were significantly affected by stage of lactation (P < 0.01). Casein as a proportion of total milk nitrogen reached a maximum in mid lactation. The proportion of alpha s- and kappa-casein in total casein decreased and the proportion of beta-casein increased systematically during lactation while the proportion of gamma-casein was lowest in mid lactation. The alpha-lactalbumin content of milk and its proportion of total whey proteins decreased during lactation. Renneting time was highest and curd firmness lowest in mid lactation. These results appeared to reflect a low degree of proteolysis in late-lactation milks compared with several other investigations, probably because of the good nutritional state of the cows. The body condition at calving affected proteolysis and the renneting properties of milk. A good body condition increased the content of whey protein in total milk nitrogen and of gamma-casein in total caseins (P < 0.05); in addition, curd firmness was improved (P < 0.01) and aggregation time was reduced (P < 0.05). We suggest that these effects were related to the fat metabolism and energy status of the cows during lactation. The interrelationships between the milk characteristics were evaluated by factor analysis to support the interpretation.

Alleles↗

In vivo identification and quantitative evaluation of carrier-mediated transport of lactate at the cellular level in the striatum of conscious, freely moving rats.

Intracerebral dialysis has allowed the continuous, on-line measurement of lactate in the extracellular fluid (ECF) of conscious, freely moving rats. The rapid time response of the technique allows the direct determination of the time course of changes in lactate in ECF following externally imposed stimuli. The time course of lactate appearance in ECF was found to be considerably slower than that observed in tissue following electroconvulsive shock or during ischemia following cardiac arrest. The ECF data could be fit to an integrated Michaelis-Menten model that assumed reversible transport of lactate across the cell membrane. This transport was found to act only when energy supplies could maintain membrane integrity and function, since ECF levels of lactate failed to follow tissue levels after cardiac arrest when energy resources are depleted. The calculated rate of cellular lactate transport was two orders of magnitude faster than transport of lactate across the blood-brain barrier in the adult rat, and passive diffusion of lactate was not found to contribute significantly across either cell or blood-brain barriers. Probenecid, an inhibitor of acid transport, was able to block both the efflux of lactate from cell to ECF and the consequent reuptake of lactate by cells in the striatum of the rat following electroconvulsive shock or ischemia.

Animals↗

Zinc needs and homeostasis during lactation.

During lactation there is an increased maternal loss of essential trace element zinc that is secreted into milk. During the first six months of lactation a mean of approximately 1.1 mg d-1 of zinc is secreted into human milk, which decreases to 0.6 mg d-1 during the next six months of lactation. The increased maternal need for zinc must be met through an increased dietary intake or homeostatic mechanisms which could compensate for the secretion of zinc into milk. These homeostatic mechanisms may include an increase in absorption, reduced excretion (urine and faecal endogenous losses) and the use of maternal pools of zinc, such as bone. Enhanced zinc absorption during lactation has been reported for lactating women whose intake of zinc is less than half of the current recommendation. Urinary zinc excretion by lactating women has also been observed to be significantly decreased up to 6 months postpartum compared to women who have never been pregnant. Approximately 30% of total body zinc is associated with bone. During lactation maternal bone resorption and reduction in one mineral content has been observed. Since urinary zinc excretion is reduced during lactation, this bone resorption could supply a portion of the zinc that is incorporated into milk. Thus, during lactation, homeostatic mechanisms which include an enhanced zinc absorption, reduced urinary zinc excretion and zinc from bone resorption could partially compensate for the secretion of zinc into human milk. These homeostatic mechanisms need to be considered when dietary recommendations for zinc intake are made for lactating women.

Absorption↗

Differential regulation of expression of genes encoding uncoupling proteins 2 and 3 in brown adipose tissue during lactation in mice.

Thermogenic activity in brown adipose tissue (BAT) decreases during lactation; the down-regulation of the gene encoding uncoupling protein 1 (UCP1) is involved in this process. Our studies show that UCP2 mRNA expression does not change during the breeding cycle in mice. In contrast, UCP3 mRNA is down-regulated in lactation but it recovers after weaning, in parallel with UCP1 mRNA. This leads to a decrease in the content of UCP3 in BAT mitochondria during lactation. Lowering the energy-sparing necessities of lactating dams by decreasing litter size or feeding with a high-fat diet prevented the down-regulation of UCP1 mRNA and UCP3 mRNA. In most cases this resulted in a less marked decrease in UCP1 and UCP3 protein in BAT mitochondria owing to lactation. Fasting for 24 h caused a different response in UCP1 and UCP3 mRNA expression: it decreased UCP1 mRNA levels but had no effect on UCP3 mRNA abundance in virgin mice; it even increased UCP3 mRNA expression in lactating dams. These changes did not lead to modifications in UCP1 or UCP3 protein abundance. Whereas acute treatment with peroxisome-proliferator-activated receptor (PPAR)alpha and PPARgamma agonists increased UCP1 mRNA levels only in lactating dams, UCP3 mRNA expression was induced by both kinds of PPAR activator in lactating dams and by PPARalpha agonists in virgin mice. It is concluded that modifications of UCP2 mRNA levels are not part of the physiological adaptations taking place in BAT during lactation. In contrast, the down-regulation of UCP3 mRNA expression and mitochondrial UCP3 content is consistent with a role for the gene encoding UCP3 in the decrease in metabolic fuel oxidation and thermogenesis in BAT during lactation.

Adipose Tissue, Brown↗

The Meyerhof quotient and the synthesis of glycogen from lactate in frog and rabbit muscle.

1. The conversion of lactate into glycogen was demonstrated in frog sartorius muscle in oxygen. The rates and amounts are highest when lactate is added to the bathing medium and are dependent on lactate and CO(2) concentration, as well as pH. The glycogen content of a resting muscle can be doubled in 4h at 24 degrees C. 2. Sartorius muscle, recovering aerobically in liquid paraffin from a period of anoxia, converts preformed lactate into glycogen at a lower rate and in smaller amounts than when lactate is added in an aqueous medium. The lower rates are similar to those Meyerhof found under the same conditions, after correction for temperature; they can be attributed partly to low muscle pH and partly to the limited amounts of lactate present. 3. Rabbit psoas muscle also shows the ability to convert added lactate into glycogen under aerobic conditions. The rates are low and similar to those in frog sartorius muscle recovering from anoxia. 4. The present experiments yield a Meyerhof quotient of 6.2, compared with Meyerhof's value of 4-5. However, these values are not significantly different from one another. 5. It is suggested that the glycogen coefficient, i.e. mol of glycogen formed/mol of lactate disappearing, is a more reliable way of assessing the resynthetic mechanism than the original quotient, i.e. mol of lactate disappearing/mol of lactate oxidized. The found coefficient is 0.419+/-0.024.

Animals↗

L-lactate transport in Ehrlich ascites-tumour cells.

Ehrlich ascites-tumour cells were investigated with regard to their stability to transport L-lactate by measuring either the distribution of [14C]lactate or concomitant H+ ion movements. The movement of lactate was dependent on the pH difference across the cell membrane and was electroneutral, as evidenced by an observed 1:1 antiport for OH- ions or 1:1 symport with H+ ions. 2. Kinetic experiments showed that lactate transport was saturable, with an apparent Km of approx. 4.68 mM and a Vmax. as high as 680 nmol/min per mg of protein at pH 6.2 and 37 degrees C. 3. Lactate transport exhibited a high temperature dependence (activation energy = 139 kJ/mol). 4. Lactate transport was inhibited competitively by (a) a variety of other substituted monocarboxylic acids (e.g. pyruvate, Ki = 6.3 mM), which were themselves transported, (b) the non-transportable analogues alpha-cyano-4-hydroxycinnamate (Ki = 0.5 mM), alpha-cyano-3-hydroxycinnamate (Ki = 2mM) and DL-p-hydroxyphenyl-lactate (Ki = 3.6 mM) and (c) the thiol-group reagent mersalyl (Ki = 125 muM). 5. Transport of simple monocarboxylic acids, including acetate and propionate, was insensitive to these inhibitors; they presumably cross the membrane by means of a different mechanism. 6. Experiments using saturating amounts of mersalyl as an "inhibitor stop" allowed measurements of the initial rates of net influx and of net efflux of [14C]lactate. Influx and efflux of lactate were judged to be symmetrical reactions in that they exhibited similar concentration dependence. 7. It is concluded that lactate transport in Ehrlich ascites-tumour cells is mediated by a carrier capable of transporting a number of other substituted monocarboxylic acids, but not unsubstituted short-chain aliphatic acids.

Animals↗