Activities Of Key Mitochondrial Enzymes Research Articles

Mitochondrial Enzyme Activities and Body Condition of Naturally Infected Sunfish (Lepomis gibbosus).

AbstractParasites can affect host behavior, cognition, locomotion, body condition, and many other physiological traits. Changes to host aerobic metabolism may be responsible for these parasite-induced performance alterations. Whole-organism metabolic rate is underpinned by cellular energy metabolism driven most prominently by mitochondria. However, few studies have explored how mitochondrial enzymatic activity relates to body condition and parasite infection, despite it being a putative site for metabolic disruptions related to health status. We studied correlations among natural parasite infection, host body condition, and activity of key mitochondrial enzymes in target organs from wild-caught pumpkinseed sunfish (Lepomis gibbosus) to better understand the cellular responses of fish hosts to endoparasite infection. Enzymatic activities in the gills, spleen, and brain of infected fish were not significantly related to parasite infection or host body condition. However, the activity of cytochrome c oxidase, an enzyme involved in oxidative phosphorylation, in fish hearts was higher in individuals with a lower body condition. Activities of citrate synthase, electron transport system (complexes I and III), and carnitine palmitoyltransferase were also significantly different among organ types. These results provide preliminary information regarding the likely mitochondrial pathways affecting host body condition, the maintenance energetic requirements of different organs, and the organs' specific dependency on particular mitochondrial pathways. These results help pave the way for future studies on the effects of parasite infection on mitochondrial metabolism.

Features of Bio Energetic Metabolism in Children Born after Assisted Reproductive Technologies

The features of the functioning of key enzymes of bio energetic metabolism in children born with the application of ART at different age periods. A cytochemical analysis of the activity of key mitochondrial enzymes was performed: succinate dehydrogenase (SDH), α- lycerophosphate dehydrogenase and α-GPDH. The material for the analysis was the buccal epithelium. The material was collected by scraping the epithelium from the inner surface of the cheek. The examined children were representatively distributed into age groups: 1st group - children of the first year old (38 children of the main and 20 children of the control group), 2nd group - from 1 to 3 years old (52 children of the main group and 20 children of the control group), 3rd group - from 4 to 7 years old (46 children of the main group and 20 children of the control group). The average age was 2.7 ± 1.9 years. Conclusion: The study of the activity of mitochondrial enzymes SDH and α-GPDH revealed the most pronounced deviations in the functioning of SDH in the group of children from 1 to 3 years old, which was associated with delayed physical development, allergic diseases, prematurity and diseases of the cardiovascular system. A decrease in the concentration of the enzyme GPDH was most expressed in the older group of children from 4 to 7 years old, and was associated with delayed physical development, Broncho-pulmonary diseases, prematurity, and diseases of the digestive system.

The adaptive response to iron involves changes in energetic strategies in the pathogen Candida albicans.

Candida albicans is an opportunist pathogen responsible for a large spectrum of infections, from superficial mycosis to systemic diseases known as candidiasis. Its ability to grow in different morphological forms, such as yeasts or filamentous hyphae, contributes to its survival in diverse microenvironments. Iron uptake has been associated with virulence, and C. albicans has developed elaborate strategies for acquiring iron from its host. In this work, we analyze the metabolic changes in response to changes in iron content in the growth medium and compare C. albicans adaptation to the presence or absence of iron. Functional and morphological studies, correlated to a quantitative proteomic analysis, were performed to assess the specific pathways underlying the response to iron, both in the yeast and filamentous forms. Overall, the results show that the adaptive response to iron is associated with a metabolic remodeling affecting the energetic pathways of the pathogen. This includes changes in the thiol‐dependent redox status, the activity of key mitochondrial enzymes and the respiratory chain. Iron deficiency stimulates bioenergetic pathways, whereas iron‐rich condition is associated with greater biosynthetic needs, particularly in filamentous forms. Moreover, we found that C. albicans yeast cells have an extraordinary capability to adapt to changes in environmental conditions.

From Africa to Antarctica: Exploring the Metabolism of Fish Heart Mitochondria Across a Wide Thermal Range.

The thermal sensitivity of ectotherms is largely dictated by the impact of temperature on cellular bioenergetics, particularly on mitochondrial functions. As the thermal sensitivity of bioenergetic pathways depends on the structural and kinetic properties of its component enzymes, optimization of their collective function to different thermal niches is expected to have occurred through selection. In the present study, we sought to characterize mitochondrial phenotypic adjustments to thermal niches in eight ray-finned fish species occupying a wide range of thermal habitats by comparing the activities of key mitochondrial enzymes in their hearts. We measured the activity of four enzymes that control substrate entrance into the tricarboxylic acid (TCA) cycle: pyruvate kinase (PK), pyruvate dehydrogenase complex (PDHc), carnitine palmitoyltransferase (CPT), and hydroxyacyl-CoA dehydrogenase (HOAD). We also assayed enzymes of the electron transport system (ETS): complexes I, II, I + III, and IV. Enzymes were assayed at five temperatures (5, 10, 15, 20, and 25°C). Our results showed that the activity of CPT, a gatekeeper of the fatty acid pathway, was higher in the cold-water fish than in the warmer-adapted fish relative to the ETS (complexes I and III) when measured close to the species optimal temperatures. The activity of HOAD showed a similar pattern relative to CI + III and thermal environment. By contrast, PDHc and PK did not show the similar patterns with respect to CI + III and temperature. Cold-adapted species had high CIV activities compared to those of upstream complexes (I, II, I + III) whereas the converse was true for warm-adapted species. Our findings reveal a significant variability of heart mitochondrial organization among species that can be linked to temperature adaptation. Cold-adapted fish do not appear to compensate for PDHc activity but likely adjust fatty acids oxidation through higher activities of CPT and HOAD relative to complexes I + III.

Short-term muscle disuse atrophy is not associated with increased intramuscular lipid deposition or a decline in the maximal activity of key mitochondrial enzymes in young and older males

Aging is generally accompanied by a progressive loss of skeletal muscle mass and impairments in metabolic function. Even a few days of muscle disuse (such as that occurring during injury or illness) leads to considerable loss of muscle mass and strength. It has been speculated that short, successive periods of muscle disuse throughout the lifespan may be largely responsible for the age-related loss of muscle mass. However, it remains unknown whether such short periods of disuse also induce impairments in metabolic function within skeletal muscle. Here, we investigated the effects of a five day period of muscle disuse on intramyocellular triacylglycerol (IMTG) content, muscle oxidative capacity, and the expression of key genes that regulate oxidative metabolism in healthy young and elderly men. Muscle biopsies were collected from healthy, young (n=12; 23±1y) and elderly (n=12; 70±1y) men prior to and immediately after a five day period of one-legged knee immobilization by way of a full leg cast. At baseline, elderly men had a greater IMTG content when compared with the young (56.2±5.1 and 34.8±7.3μmol·g−1, respectively; P<0.05) with no changes in either group following immobilization (53.4±5.0 and 35.7±5.0μmol·g−1, respectively; P>0.05). In line, five days of disuse did not lower citrate synthase, β-HAD or cytochrome C oxidase activity in skeletal muscle tissue. Pyruvate dehydrogenase activity increased following immobilization in the older subjects only, from 0.39±0.06 to 0.55 0.05μmol·g−1·min−1 (71±33%; P<0.01). The skeletal muscle mRNA expression of PGC1α and citrate synthase both declined following immobilization in both the young and elderly subjects. We conclude that five days of muscle disuse does not increase intramuscular lipid deposition or reduce the maximal activity of key mitochondrial enzymes within the skeletal muscle of young or older men.

Elevated temperature and PCO2 shift metabolic pathways in differentially oxidative tissues of Notothenia rossii

Mitochondrial plasticity plays a central role in setting the capacity for acclimation of aerobic metabolism in ectotherms in response to environmental changes. We still lack a clear picture if and to what extent the energy metabolism and mitochondrial enzymes of Antarctic fish can compensate for changing temperatures or PCO2 and whether capacities for compensation differ between tissues. We therefore measured activities of key mitochondrial enzymes (citrate synthase (CS), cytochrome c oxidase (COX)) from heart, red muscle, white muscle and liver in the Antarctic fish Notothenia rossii after warm- (7°C) and hypercapnia- (0.2kPa CO2) acclimation vs. control conditions (1°C, 0.04kPa CO2). In heart, enzymes showed elevated activities after cold-hypercapnia acclimation, and a warm-acclimation-induced upward shift in thermal optima. The strongest increase in enzyme activities in response to hypercapnia occurred in red muscle. In white muscle, enzyme activities were temperature-compensated. CS activity in liver decreased after warm-normocapnia acclimation (temperature-compensation), while COX activities were lower after cold- and warm-hypercapnia exposure, but increased after warm-normocapnia acclimation. In conclusion, warm-acclimated N. rossii display low thermal compensation in response to rising energy demand in highly aerobic tissues, such as heart and red muscle. Chronic environmental hypercapnia elicits increased enzyme activities in these tissues, possibly to compensate for an elevated energy demand for acid-base regulation or a compromised mitochondrial metabolism, that is predicted to occur in response to hypercapnia exposure. This might be supported by enhanced metabolisation of liver energy stores. These patterns reflect a limited capacity of N. rossii to reorganise energy metabolism in response to rising temperature and PCO2.

The curious case of anabolic resistance: old wives' tales or new fables?

the progressive loss of muscle mass, commonly termed sarcopenia, accompanies the process of healthy aging ([27][1]). The underlying basis of this condition, in a very simplistic view, would be at a more advanced age skeletal muscle proteins are being lost because of an imbalance between muscle

Protective effects of compound FLZ, a novel synthetic analogue of squamosamide, on β-amyloid-induced rat brain mitochondrial dysfunction in vitro

The aim of the present study was to assess the effects of N-[2-(4-hydroxyphenyl)ethyl]-2-(2,5-dimethoxyphenyl)-3-(3-methoxy-4-hydroxyphenyl) acrylamide (compound FLZ), a novel synthetic analogue of squamosamide, on the dysfunction of rat brain mitochondria induced by Abeta(25-35) in vitro. Isolated rat brain mitochondria were incubated with aged Abeta(25-35) for 30 min in the presence and absence of FLZ (1-100 micromol/L). The activities of key mitochondrial enzymes, the production of hydrogen peroxide (H(2)O(2)) and superoxide anion (O2*-), and the levels of glutathione (GSH) in mitochondria were examined. Mitochondrial swelling and the release of cytochrome c from mitochondria were assessed by biochemical and Western blot methods, respectively. Incubation of mitochondria with aged Abeta(25-35) inhibited the activities of alpha-ketoglutarate dehydrogenase (alpha-KGDH), pyruvate dehydrogenase (PDH) and respiratory chain complex IV. It also resulted in increased H(2)O(2) and (O2*-) production, and decreased the GSH level in mitochondria. Furthermore, it induced mitochondrial swelling and cytochrome c release from the mitochondria. The addition of FLZ (100 micromol/L) prior to treatment with Abeta(25-35) significantly prevented these toxic effects of Abeta(25-35) on the mitochondria. FLZ has a protective effect against Abeta(25-35)-induced mitochondrial dysfunction in vitro.

Calorie Restriction Increases Muscle Mitochondrial Biogenesis in Healthy Humans

BackgroundCaloric restriction without malnutrition extends life span in a range of organisms including insects and mammals and lowers free radical production by the mitochondria. However, the mechanism responsible for this adaptation are poorly understood.Methods and FindingsThe current study was undertaken to examine muscle mitochondrial bioenergetics in response to caloric restriction alone or in combination with exercise in 36 young (36.8 ± 1.0 y), overweight (body mass index, 27.8 ± 0.7 kg/m2) individuals randomized into one of three groups for a 6-mo intervention: Control, 100% of energy requirements; CR, 25% caloric restriction; and CREX, caloric restriction with exercise (CREX), 12.5% CR + 12.5% increased energy expenditure (EE). In the controls, 24-h EE was unchanged, but in CR and CREX it was significantly reduced from baseline even after adjustment for the loss of metabolic mass (CR, −135 ± 42 kcal/d, p = 0.002 and CREX, −117 ± 52 kcal/d, p = 0.008). Participants in the CR and CREX groups had increased expression of genes encoding proteins involved in mitochondrial function such as PPARGC1A, TFAM, eNOS, SIRT1, and PARL (all, p < 0.05). In parallel, mitochondrial DNA content increased by 35% ± 5% in the CR group (p = 0.005) and 21% ± 4% in the CREX group (p < 0.004), with no change in the control group (2% ± 2%). However, the activity of key mitochondrial enzymes of the TCA (tricarboxylic acid) cycle (citrate synthase), beta-oxidation (beta-hydroxyacyl-CoA dehydrogenase), and electron transport chain (cytochrome C oxidase II) was unchanged. DNA damage was reduced from baseline in the CR (−0.56 ± 0.11 arbitrary units, p = 0.003) and CREX (−0.45 ± 0.12 arbitrary units, p = 0.011), but not in the controls. In primary cultures of human myotubes, a nitric oxide donor (mimicking eNOS signaling) induced mitochondrial biogenesis but failed to induce SIRT1 protein expression, suggesting that additional factors may regulate SIRT1 content during CR.ConclusionsThe observed increase in muscle mitochondrial DNA in association with a decrease in whole body oxygen consumption and DNA damage suggests that caloric restriction improves mitochondrial function in young non-obese adults.

Abstracts for Oral Presentation

Abstracts for Oral Presentation

Aerobic capacity of frog skeletal muscle during hibernation.

Frogs submerged at 3 degrees C in hypoxic water (Po2=60 mmHg) depress their metabolic rate to 25% of that seen in control animals with access to air. The hypometabolic state of the skeletal muscle in such cold-submerged frogs is thought to be the most important contributor to the overall metabolic depression. The aim of this study was to determine whether the aerobic capacity of frog skeletal muscle became altered during 1-4 mo of hibernation to match the reduction in adenosine triphosphate (ATP) demand. To this end, the activities of key mitochondrial enzymes were measured in the skeletal muscle and in isolated mitochondria of frogs at different stages during hibernation. We also measured the activity of lactate dehydrogenase (LDH) as an indicator of glycolytic capacity. The activities of cytochrome c oxidase, citrate synthase, and LDH were significantly lower in frog skeletal muscle after 4 mo of hibernation compared with control conditions. The reduction in skeletal muscle aerobic capacity is apparently due to changes in the intrinsic properties of the mitochondria. Overall, these results indicate an important reorganisation of ATP-producing pathways during long-term metabolic depression to match the lowered ATP demand.

Elevated intramyocellular lipid concentration in obese subjects is not reduced after diet and exercise training.

To determine the effects of weight loss on intramyocellular energy substrates, vastus lateralis muscle biopsies were taken from six obese subjects (body mass index 34 +/- 5 kg/m(2)) before, after 15 wk of energy restriction (ER; -700 kcal/day), and after a further average 20.7 +/- 1.6 wk of endurance training plus low-fat diet (ET-LFD). Body weight fell from 100 +/- 6 to 89 +/- 6 kg during ER and to 84 +/- 4 kg after ET-LFD. Lipids and glycogen were histochemically measured in type I, IIA, and IIB fibers. Total muscle glycogen content (MGC; per 100 fibers) decreased after ER [from 72 +/- 13 to 55 +/- 8 arbitrary units (AU)]. A similar but not significant decrease was seen in total muscle lipid content (MLC; 14 +/- 5 to 9 +/- 1 AU). After ET-LFD, MGC returned to initial values (74 +/- 8 AU), and MLC approached near-initial values (12 +/- 3 AU). Individual fiber lipid concentration did not change throughout the protocol in all fiber types, whereas glycogen concentration increased after ET-LFD. The training effects of ET-LFD were measured as increasing activities of key mitochondrial enzymes. Although total muscle energy reserves can be reduced after weight loss, their concentration within individual myofibers remains elevated. Weight loss does not appear sufficient to correct the potential detrimental effects of high intracellular lipid concentrations.

Effects of outlet obstruction and its reversal on mitochondrial enzyme activity in rabbit urinary bladders.

This study investigated the effects of outlet obstruction and its reversal on mitochondrial enzyme activity in the rabbit urinary bladder. We induced mild bladder outlet obstruction in male New Zealand White rabbits. Following two weeks of obstruction, one group of animals (n = 6) was sacrificed, while outlet obstruction was relieved in three additional groups of animals, which were sacrificed one (n = 5), two (n = 5) and four (n = 5) weeks after relieving the obstruction. Seven sham operated rabbits served as controls. We extracted mitochondria from fresh detrusor and assayed activities of key mitochondrial enzymes in the citric acid cycle, citrate synthase and malate dehydrogenase, as well as those in the electron transport chain, succinate cytochrome c reductase, NADH-cytochrome c reductase and cytochrome c oxidase. With high performance liquid chromatography (HPLC) we determined the tissue content of phosphocreatine and the adenine nucleotides (ATP, ADP and AMP), which was used for calculating energy charge. Responses of detrusor strips to 500 microM bethanechol and 120 mM KCl provided the assessment of detrusor contractility. Contractile response of the detrusor strips to bethanechol stimulation was significantly reduced by outlet obstruction, nevertheless, it recovered gradually toward the control level after the relief of outlet obstruction. Outlet obstruction reduced the detrusor content of phosphocreatine, ATP and energy charge. After relieving the obstruction, however, these recovered gradually, reaching control levels 4 weeks later. The activities of all assayed enzymes were reduced by two weeks of outlet obstruction. Relieving the obstruction restored enzyme activity gradually but at different rates for different enzymes. Activities of the citric acid cycle enzymes citrate synthase and malate dehydrogenase recovered and were similar to control levels four weeks after relief of the obstruction. Of the enzymes in the electron transport chain, NADH cytochrome reductase activity recovered most quickly by one week after relief of the obstruction. The activity of cytochrome c oxidase improved more slowly, but 4 weeks after relieving the obstruction it, also, was restored and was similar to the control level. Succinate cytochrome reductase activity remained lower than the control over the entire four weeks of recovery. The close association between mitochondrial enzyme activity, energy metabolism and contractility of the detrusor indicates the important role of mitochondrial enzyme damage in decreasing energy production and impairing contractile function of the urinary bladders following outlet obstruction. Our findings also show that various mitochondrial enzymes exhibit different susceptibilities and reversibilities to pathological stress.

EFFECTS OF OUTLET OBSTRUCTION AND ITS REVERSAL ON MITOCHONDRIAL ENZYME ACTIVITY IN RABBIT URINARY BLADDERS

Purpose This study investigated the effects of outlet obstruction and its reversal on mitochondrial enzyme activity in the rabbit urinary bladder. Materials and Methods We induced mild bladder outlet obstruction in male New Zealand White rabbits. Following two weeks of obstruction, one group of animals (n = 6) was sacrificed, while outlet obstruction was relieved in three additional groups of animals, which were sacrificed one (n = 5), two (n = 5) and four (n = 5) weeks after relieving the obstruction. Seven sham operated rabbits served as controls. We extracted mitochondria from fresh detrusor and assayed activities of key mitochondrial enzymes in the citric acid cycle, citrate synthase and malate dehydrogenase, as well as those in the electron transport chain, succinate cytochrome c reductase, NADH-cytochrome c reductase and cytochrome c oxidase. With high performance liquid chromatography (HPLC) we determined the tissue content of phosphocreatine and the adenine nucleotides (ATP, ADP and AMP), which was used for calculating energy charge. Responses of detrusor strips to 500 micro M bethanechol and 120 mM KCl provided the assessment of detrusor contractility. Results Contractile response of the detrusor strips to bethanechol stimulation was significantly reduced by outlet obstruction, nevertheless, it recovered gradually toward the control level after the relief of outlet obstruction. Outlet obstruction reduced the detrusor content of phosphocreatine, ATP and energy charge. After relieving the obstruction, however, these recovered gradually, reaching control levels 4 weeks later. The activities of all assayed enzymes were reduced by two weeks of outlet obstruction. Relieving the obstruction restored enzyme activity gradually but at different rates for different enzymes. Activities of the citric acid cycle enzymes citrate synthase and malate dehydrogenase recovered and were similar to control levels four weeks after relief of the obstruction. Of the enzymes in the electron transport chain, NADH cytochrome reductase activity recovered most quickly by one week after relief of the obstruction. The activity of cytochrome c oxidase improved more slowly, but 4 weeks after relieving the obstruction it, also, was restored and was similar to the control level. Succinate cytochrome reductase activity remained lower than the control over the entire four weeks of recovery. Conclusions The close association between mitochondrial enzyme activity, energy metabolism and contractility of the detrusor indicates the important role of mitochondrial enzyme damage in decreasing energy production and impairing contractile function of the urinary bladders following outlet obstruction. Our findings also show that various mitochondrial enzymes exhibit different susceptibilities and reversibilities to pathological stress.