It is a well-documented finding that blood lactate concentrations at any given absolute or relative (% of maximum oxygen uptake; % VO₂ₘₐₓ) workload, are lower following endurance training. The search for the mechanisms responsible for lower blood lactate concentrations after training, however, has led to conflicting results, particularly when the possible causes of this finding have been investigated in humans. In this study, three questions related to the effects of endurance training on lactate metabolism were investigated.

The effects of an increase in body temperature per se on the lactate and pyruvate concentrations of the arterial blood, muscle venous blood, sagittal sinus blood, and cerebrospinal fluid were determined. Paralysed anesthetized dogs with near normal arterial pH and PC0(2) values were ventilated with a 50% 0(2), 50% N(2) mixture and heated to a temperature of 42°C and maintained at this temperature for a period of 2 hours. During hyperthermia a slight increase in lactate and pyruvate was observed in the arterial blood. However, this was not statistically significant. Also a slight increase in the concentration of these substances occurred in the muscle venous blood and sagittal sinus blood. This change, however, was parallel to that observed in the arterial blood. The lactate-pyruvate ratio of the arterial blood, muscle venous blood and sagittal sinus blood did not show any significant change and thus no increase in anaerobic processes was detected during the hyperthermic period. On the other hand the cerebrospinal fluid lactate and pyruvate increased significantly throughout the hyperthermic period but maintained a constant lactate-pyruvate ratio. The results indicate that the increase of lactate and pyruvate in the cerebrospinal fluid are a result of an increased rate of aerobic glycolysis. The fact that the increases observed in the cerebrospinal fluid lactate and pyruvate were not reflected in the cerebral venous blood indicates lactate and pyruvate may have difficulty in diffusing across the blood brain barrier and cerebral venous blood is thus a poor index of cerebral lactate and pyruvate changes. / Medicine, Faculty of / Cellular and Physiological Sciences, Department of / Graduate

During sustained exercise, working muscles must be supplied with adequate kinds and amounts of exogenous fuels, and the delivery rates of oxygen and oxidizable substrates should be matched. The study of metabolite fluxes and their regulation is therefore critical to the understanding of exercise metabolism. Lactate has received renewed attention from physiologists and biochemists with the realization that it is not only an end product of glycolysis, but also an important fuel for aerobic work. As an oxidizable fuel, this substrate may provide some performance advantage over other fuels such as glucose and free fatty acids. The goals of this thesis were: 1) to determine whether endurance-adapted animals can support higher plasma lactate turnover rates than sedentary animals; and 2) to investigate the major factors involved in the regulation of plasma metabolite turnover at the whole-organism level - using lactate as a model. Lactate turnover rates were measured by bolus injection of [U-¹⁴C]lactate in skipjack tuna, Katsuwonus pelamis, and in thoroughbred racehorses, Equus caballus. In tuna, turnover rates ranged from 112 to 431 umol min⁻¹ kg⁻¹ and they were positively correlated with lactate concentration (slope = 15.1, r = 0.92). This teleost is able to support higher plasma lactate turnover rates than expected for a mammalian lower temperature, and lactate is probably an important oxidizable fuel in this species. For comparative purposes, resting turnover rates of lactate and glucose were plotted versus body mass on a log-log scale for a wide range of mammalian species. These plots were linear, and they showed the same slope as the classic body mass vs metabolic rate relationship. Thoroughbred horses are likely to have an aerobic scope of 40-fold or more. One of their main physiological adaptations to exercise is the ability to increase hematocrit by more than one and a half-fold in response to exercise. In the present study, this adjustment allowed them to reach an A-V difference in 0₂ content of more than 23 vol% during maximal exercise, a much higher value than other mammals. Their lactate turnover rate and cardiac output were measured at rest and two levels of submaximal exercise (45 and 55 V0₂ max) to investigate the relationship between cardiovascular adjustments on plasma lactate turnover rate. Cardiac output ranged from 106 to 571 ml min⁻¹ kg⁻¹, and mean lactate turnover rate from 9.3 at rest, to 75.9 umol min⁻¹ kg⁻¹ at 55% V0₂ max. In contrast with the situation found in tuna, the lactate turnover rates of thoroughbreds were not elevated compared with other mammals, showing that the metabolic adaptations of these outstanding athletes do not include the ability to sustain higher lactate fluxes than sedentary animals. In horses, the contribution of plasma lactate oxidation to V0₂ is minimal, and this substrate is not an important oxidative fuel; lipid oxidation may represent their major pathway for aerobic energy production during exercise. The ability to oxidize plasma lactate at high rates is therefore not necessarily required for the "elite" performance of endurance exercise. This study also shows that both, plasma lactate concentration and cardiac output are positively correlated with turnover rate. The correlation between cardiac output and lactate turnover rate is independent of the relationship between plasma lactate concentration and turnover rate. Plasma metabolite concentration and cardiac output can be regulators of plasma metabolite turnover rate. It is proposed that these two variables are, respectively, the fine and coarse controls for flux rate adjustments during exercise. / Science, Faculty of / Zoology, Department of / Graduate

This study examines factors that determine blood lactate accumulation during exercise in order to examine the opposing theories that such accumulation occurs either as a result of muscle anaerobiosis or an "overflow" of oxygen-independent glycolysis.

Aging is a multifaceted process of accumulation of damage and waste in cells and tissues; age-related changes in mitochondria and in respiratory metabolism have the focus of aging research for decades. Studies of aging in nematodes, flies and mammals all revealed age-related decline in respiratory functions, with somewhat controversial causative role. Here we investigated age-related changes in respiration rates, lactate/pyruvate ratio, a commonly used proxy for NADH/NAD+ balance, and mitochondrial membrane potential in 4 genotypes of an emerging model organism for aging research, a cyclic parthenogen Daphnia magna. We show that total body weight-adjusted respiration rate decreased with age, although this decrease was small in magnitude and could be fully accounted for by the decrease in locomotion and feeding activity. Neither total respiration normalized by protein content, nor basal respiration rate measured in anaesthetized animals decreased with age. Lactate/pyruvate ratio and mitochondrial membrane potential (∆Ψ) showed no age-related changes, with possible exceptions of ∆Ψ in epipodites (excretory and gas exchange organs) in which ∆Ψ decreased with age and in the optical lobe of the brain, in which ∆Ψ showed a maximum at middle age. We conclude that actuarial senescence in Daphnia is not caused by a decline in respiratory metabolism and discuss possible mechanisms of maintaining mitochondrial healthspan throughout the lifespan.

Circulating lactate levels are a critical biomarker for sepsis and are positively correlated with sepsis-associated mortality. We investigated whether lactate plays a biological role in causing endothelial barrier dysfunction in sepsis. We showed that lactate causes vascular permeability and worsens organ dysfunction in CLP sepsis. Mechanistically, lactate induces ERK-dependent activation of calpain1/2 for VE-cadherin proteolytic cleavage, leading to the enhanced endocytosis of VE-cadherin in endothelial cells. In addition, we found that ERK2 interacts with VE-cadherin and stabilizes VE-cadherin complex in resting endothelial cells. Lactate-induced ERK2 phosphorylation promotes ERK2 disassociation from VE-cadherin. In vivo suppression of lactate production or genetic depletion of lactate receptor GPR81 mitigates vascular permeability and multiple organ injury and improves survival outcome in polymicrobial sepsis. Our study reveals that metabolic cross-talk between glycolysis-derived lactate and the endothelium plays a critical role in the pathophysiology of sepsis.