Biomedical subjects

Christopher D Moyes

Publications and source records attributed to Christopher D Moyes.

12 recordsLinked to original sources

PubMed2006

Molecular evolution of cytochrome c oxidase in high-performance fish (teleostei: Scombroidei).

The 13 peptides encoded by vertebrate mitochondrial DNA (mtDNA) are essential subunits of oxidative phosphorylation (OXPHOS) enzymes. These genes normally experience purifying selection and also coevolve with nuclear-encoded subunits of OXPHOS complexes. However, the role of positive selection on mtDNA evolution is still unclear, as most examples of intergenomic coevolution appear to be the result of compensation by nuclear-encoded genes for mildly deleterious mtDNA mutations, and not simultaneous positive selection in both genomes. Organisms that have experienced strong selective pressures to increase aerobic capacity or adapt to changes in thermal environment may be better candidates in which to examine the impact of positively selected changes on mtDNA evolution. The tuna (suborder Scombroidei, family Scombridae) and billfish (suborder Scombroidei, families Xiphiidae and Istiophoridae) are highly aerobic fish with multiple specializations in muscle energetics, including a high mitochondrial content and regional endothermy. We examined the role of positively selected mtDNA substitutions in the production of these unique phenotypes. Focusing on a catalytic subunit of cytochrome c oxidase (COX II), we found that the rate ratio of nonsynonymous (d(N); amino acid changing)-to-synonymous (d(S); silent) substitutions was not increased in lineages leading to the tuna but was significantly increased in the lineage preceding the billfish. Furthermore, there are a number of individual positively selected sites that, when mapped onto the COX crystal structure, appear to interact with other COX subunits and may affect OXPHOS function and regulation in billfish.

Animals↗

PubMed2006

Regulation of skeletal muscle mitochondrial content during aging.

Mitochondrial content of skeletal muscle varies among fiber types, and changes in complex ways during aging. We evaluated the regulatory origins of differences in mitochondrial content among muscles of varied fiber type in F344xBNF1 rats, and how these regulatory patterns are altered with aging. In adult (12 month) animals we found that units citrate synthase (CS)/g tissue, a marker for mitochondrial content, varied approximately 3-fold among 10 skeletal muscles. Stoichiometric relationships between CS and isocitrate dehydrogenase, aconitase, and cytochrome c oxidase were generally preserved across fiber types. Among the 10 muscles of adult rats, CS content correlated with nuclear content (R2= 0.36). Muscles differed widely in CS messenger RNA (mRNA)/DNA (an index of variation in transcriptional regulations) and units CS/CS mRNA (an index of variation in posttranscriptional regulations). All muscles of aged rats (35 months) showed an increase in mg DNA/g, suggestive of atrophy. Age-dependent declines in units CS/DNA were accompanied by reductions in CS mRNA/DNA and/or units CS/CS mRNA, depending on muscle fiber type. Thus, declines in units CS/DNA with age appeared to be due to transcriptional as well as translational variations. Differences in mitochondrial content among muscle fiber types and age groups may arise from variations in nuclear content and posttranscriptional processes, as well as transcriptional regulation.

Aging↗

PubMed2006

Control of mitochondrial gene expression in the aging rat myocardium.

Aging induces complex changes in myocardium bioenergetic and contractile properties. Using F344BNF(1) rats, we examined age-dependent changes in myocardial bioenergetic enzymes (catalytic activities and transcript levels) and mRNA levels of putative transcriptional regulators of bioenergetic genes. Very old rats (35 months) showed a 22% increase in ventricular mass with no changes in DNA or RNA per gram. Age-dependent cardiac hypertrophy was accompanied by complex changes in mitochondrial enzymes. Enzymes of the Krebs cycle and electron transport system remained within 15% of the values measured in adult heart, significant decreases occurring in citrate synthase (10%) and aconitase (15%). Transcripts for these enzymes were largely unaffected by aging, although mRNA levels of putative transcriptional regulators of the enzymes (nuclear respiratory factor (NRF) 1 and 2 alpha subunit) increased by about 30%-50%. In contrast, enzymes of fatty acid oxidation exhibited a more diverse pattern, with a 50% decrease in beta-hydroxyacyl-CoA dehydrogenase (HOAD) and no change in long-chain acyl-CoA dehydrogenase or carnitine palmitoyltransferase. Transcript levels for fatty acid oxidizing enzymes covaried with HOAD, which declined significantly by 30%. There were no significant changes in the relative transcript levels of regulators of genes for fatty acid oxidizing enzymes: peroxisome proliferator-activated receptor-alpha (PPARalpha), PPARbeta, or PPARgamma coactivator-1alpha (PGC-1alpha). There were no changes in the mRNA levels of Sirt1, a histone-modifying enzyme that interacts with PGC-1alpha. Collectively, these data suggest that aging causes complex changes in the enzymes of myocardial energy metabolism, triggered in part by NRF-independent pathways as well as post-transcriptional regulation.

Aging↗

PubMed2005

Motility, ATP levels and metabolic enzyme activity of sperm from bluegill (Lepomis macrochirus).

Male bluegill displays one of two life history tactics. Some males (termed "parentals") delay reproduction until ca. 7 years of age, at which time they build nests and actively courts females. Others mature precociously (sneakers) and obtain fertilizations by cuckolding parental males. In the current study, we studied the relations among sperm motility, ATP levels, and metabolic enzyme activity in parental and sneaker bluegill. In both reproductive tactics, sperm swimming speed and ATP levels declined in parallel over the first 60 s of motility. Although sneaker sperm initially had higher ATP levels than parental sperm, by approximately 30 s postactivation, no differences existed between tactics. No differences were noted between tactics in swimming speed, percent motility, or the activities of key metabolic enzymes, although sperm from parentals had a higher ratio of creatine phosphokinase (CPK) to citrate synthase (CS). In both tactics, with increasing CPK and CS activity, sperm ATP levels increased at 20 s postactivation, suggesting that capacities for phosphocreatine hydrolysis and aerobic metabolism may influence interindividual variation in rates of ATP depletion. Nonetheless, there was no relation between sperm ATP levels and either swimming speed or percent of sperm that were motile. This suggests that interindividual variation in ATP levels may not be the primary determinant of variation in sperm swimming performance in bluegill.

Adenosine Triphosphate↗

PubMed2005

Control of muscle bioenergetic gene expression: implications for allometric scaling relationships of glycolytic and oxidative enzymes.

Muscle metabolic properties vary with body size, with larger animals relying relatively less on oxidative metabolism as a result of lower specific activities of mitochondrial enzymes and greater specific activities of glycolytic enzymes. While many have argued reasons why such relationships might be grounded in physical relationships, an explanation for the regulatory basis of the differences in enzyme levels remains unexplored. Focusing on skeletal muscle, we review potential cellular and genetic explanations for the relationship between bioenergetic enzymes and body mass. Differences in myonuclear domain (the ratio of fiber volume to nuclei number) in conjunction with constitutive expression may explain part of the variation in mitochondrial content among fiber types and species. Superimposed on such constitutive determinants are (1) extrinsic signalling pathways that control the muscle contractile and metabolic phenotype and (2) intrinsic signalling pathways that translate changes in cellular milieu (ions, metabolites, oxygen, redox) arising through the contractile phenotype into changes in enzyme synthesis. These signalling pathways work through transcriptional regulation, as well as post-transcriptional, translational and post-translational regulation, acting via synthesis and degradation.

Animals↗

PubMed2004

Mitochondrial enzyme content in the muscles of high-performance fish: evolution and variation among fiber types.

Muscle mitochondrial content varies widely among fiber types and species. We investigated the origins of variation in the activity of the mitochondrial enzyme citrate synthase (CS), an index of mitochondrial abundance, among fiber types and species of high-performance fish (tunas and billfishes). CS activities varied up to 30-fold among muscles: lowest in billfish white muscle and highest in billfish heater organ. Among species, CS activities of red, white, and cardiac muscles of three tuna species were twofold greater than the homologous muscles of two billfish species. Because comparisons of CS amino acid sequences deduced from a combination of PCR methods argue against clade-specific differences in catalytic properties, CS activity reflects CS content among these five species. To assess the bases of these differences in CS activity, we looked at the relationship between CS activity (U/g muscle), nuclear content (DNA/g muscle), and CS transcript levels (CS mRNA/g RNA). Muscle CS activity differed by 10- to 30-fold when expressed per gram of muscle but only threefold when expressed per milligram of DNA. Thus it is nuclear DNA content, not fiber-type differences, in CS gene expression that may be the main determinant of CS activity in muscle. Conversely, evolutionary (tunas vs. billfishes) differences in CS arise from differences in posttranscriptional regulation, based on relationships between CS enzyme levels and CS mRNA assessed by quantitative competitive RT-PCR. These data argue that fiber-type differences can arise without major differences in fiber-type-specific regulation of the CS gene, whereas evolutionary differences may be largely due to posttranscriptional regulation.

Amino Acid Sequence↗

PubMed2004

Using humans to study the physiological evolution of energy metabolism: a tribute to Peter Hochachka.

Comparative physiology traditionally relies on unusual animals to explore the mechanistic basis or evolution of physiological processes. Despite his credentials as a comparative physiologist, much of the recent work by Peter Hochachka relied upon humans as "comparative" models. Technological advances in clinical analyses and expanding databases of genomic diversity have made humans a very "convenient" model to study the evolution of metabolism. For example, a growing number of metabolic diseases have been shown to have a genetic basis. Obesity, type II diabetes, metabolic syndrome X may each have linkages to polymorphisms in genes encoding transcriptional regulators of bioenergetic genes. Genetic polymorphisms and gene families for these important transcription factors inspire questions about the evolution of energy metabolism in other animals. Thus, these biomedical studies provide important direction for researchers interested in exploring the regulatory basis of interspecies variations in bioenergetics.

Biological Evolution↗

PubMed2004

Bioenergetic remodeling during cellular differentiation: changes in cytochrome c oxidase regulation do not affect the metabolic phenotype.

Myogenesis induces mitochondrial proliferation, a decrease in reactive oxygen species (ROS) production, and an increased reliance upon oxidative phosphorylation. While muscles typically possess 20%-40% excess capacity of cytochrome c oxidase (COX), undifferentiated myoblasts have only 5%-20% of the mitochondrial content of myotubes and muscles. We used two muscle lines (C2C12, Sol8) and 3T3-L1 pre-adipocytes to examine if changes in COX regulation or activity with differentiation cause a shift in metabolic phenotype (i.e., more oxidative, less glycolytic, less ROS). COX activity in vivo can be suppressed by its inhibitor, nitric oxide, or sub-optimal substrate (cytochrome c) concentrations. Inhibition of nitric oxide synthase via L-NAME had no effect on the respiration of adherent undifferentiated cells, although it did stimulate respiration of myoblasts in suspension. While cytochrome c content increased during differentiation, there was no correlation with respiratory rate or reliance on oxidative metabolism. There was no correlation between COX specific activity and oxidative metabolism between cell type or in relation to differentiation. These studies show that, despite the very low activities of COX, undifferentiated myoblasts and pre-adipocytes possess a reserve of COX capacity and changes in COX with differentiation do not trigger the shift in metabolic phenotype.

Adipocytes↗

PubMed2003

Controlling muscle mitochondrial content.

Mitochondrial content, a chief determinant of aerobic capacity, varies widely among muscle types and species. Mitochondrial enzyme levels in vertebrate skeletal muscles vary more than 100-fold, from fish white muscle to bird flight muscles. Recent studies have shed light on the transcriptional regulators that control mitochondrial gene expression in muscle fiber differentiation and development, and in the context of pathological conditions such as neuromuscular disease and obesity. While the transcriptional co-activator PGC-1alpha (peroxisome proliferator-activated receptor gamma co-activator 1) has emerged as a master controller of mitochondrial gene expression, it is important to consider other mechanisms by which coordinated changes in mitochondrial content could arise. These studies, largely using biomedical models, provide important information for comparative biologists interested in the mechanistic basis of inter-species variation in muscle aerobic capacity.

Animals↗

PubMed2002

Origins and consequences of mitochondrial variation in vertebrate muscle.

This review addresses the mechanisms by which mitochondrial structure and function are regulated, with a focus on vertebrate muscle. We consider the adaptive remodeling that arises during physiological transitions such as differentiation, development, and contractile activity. Parallels are drawn between such phenotypic changes and the pattern of change arising over evolutionary time, as suggested by interspecies comparisons. We address the physiological and evolutionary relationships between ATP production, thermogenesis, and superoxide generation in the context of mitochondrial function. Our discussion of mitochondrial structure focuses on the regulation of membrane composition and maintenance of the three-dimensional reticulum. Current studies of mitochondrial biogenesis strive to integrate muscle functional parameters with signal transduction and molecular genetics, providing insight into the origins of variation arising between physiological states, fiber types, and species.

Animals↗

PubMed2002

Chronic treatment with azide in situ leads to an irreversible loss of cytochrome c oxidase activity via holoenzyme dissociation.

Chronic treatment of cultured cells with very low levels of azide (I(50)<10 microm) leads to slow (t(12) = 6 h), irreversible loss of cytochrome c oxidase (COX) activity. Azide-mediated COX losses were not accompanied by inhibition of other mitochondrial enzymes and were not dependent upon electron flux through oxidative phosphorylation. Although azide treatment also reduced activity (but not content) of both CuZn superoxide dismutase and catalase, a spectrum of pro-oxidants (and anti-oxidants) failed to mimic (or prevent) azide effects, arguing that losses in COX activity were not due to resultant compromises in free radical scavenging. Loss of COX activity was not attributable to reduced rates of mitochondrial protein synthesis or declines in either COX subunit mRNA or protein levels (COX I, II, IV). Co-incubation experiments using copper (CuCl(2), Cu-His) and copper chelators (neocuproine, bathocuproine) indicated that azide effects were not mediated by interactions with either Cu(A) or Cu(B). In contrast, difference spectroscopy and high performance liquid chromatography analyses demonstrated azide-induced losses in cytochrome aa(3) content although not to the same extent as catalytic activity. Differential azide effects on COX content relative to COX activity were confirmed using a refined inhibition time course in combination with blue native electrophoresis, and established that holoenzyme dissociation occurs subsequent to losses in catalytic activity. Collectively, these data suggest that COX deficiency can arise through enhanced holoenzyme dissociation, possibly through interactions with the structure or coordination of its heme moieties.

Azides↗

PubMed2002

Analysis of mucosal stress response in acid-induced esophagitis in opossum.

This study is the first to examine site-specific changes in mucosal antioxidants and expression and localization of heat shock proteins (HSPs) following the induction of subacute esophagitis and after recovery using an established animal model. Distal, middle, and proximal samples were excised from anesthetized opossums 24 hr after three consecutive days of 45-min perfusion with saline or 100 mmol/liter HCI, or seven days after acid in recovery animals. Compared to controls, acid-induced erosive esophagitis significantly increased glutathione peroxidase and HSP90 at all sites and HSP60 proximally. Reduced glutathione was significantly decreased distally, as was HSP72 at distal and middle sites. No changes in superoxide dismutase or catalase occurred. After recovery, superoxide dismutase, catalase, and HSP expression were not different from controls. Glutathione peroxidase and glutathione were significantly decreased distally. Similar differential stress responses may occur in patients with chronic gastroesophageal reflux and could be important in the pathogenesis of reflux esophagitis.

Acids↗