Thermal relations in sled dogs before and after exercise.

Regulation of internal body temperature (Tb), or thermoregulation, is an evolutionarily conserved trait that places demand on basal metabolic rate of endothermic animals. Across species, athletes generate increased quantities of heat in comparison to their nonathletic counterparts and, therefore, must mediate physiological unbalance by upregulating the effectiveness of their heat dissipation abilities. Canine athletes are no exception to this phenomenon, however, with literature denoting body temperatures lower than nonathletic canines, it is clear they must possess adaptations to mitigate this demand. With VO2 max measurements of more than 200 mL/kg/min in sled dogs with mild training to 300 mL/kg/min in highly trained animals, sled dogs are a prime example of athleticism in canines. Seeking to determine correlations between Tear and body mass, morphology, and age of canine athletes, core body temperature (Tb) was measured with an instant ear thermometer, using Tear as a correlate before and after a 2-mile run. In addition, we employed thermal imaging analysis to capture body-wide heat dissipation patterns in sled dogs, and focused on thermal variation of mouth (Tmouth), nose (Tnose), and eyes (Teye). Furthermore, we looked at correlations between thermal variability across these four tissues and head morphology of each dog. Tear was consistently the highest temperature across all tissues measured, with a 1.5°C increase between pre- to postexercise (p < 0.001). Thermal imaging revealed significant positive correlations between Tmouth and body mass 15 min postexercise (p = 0.0023) as well as significantly negative correlations between Tnose and body mass at before exercise (p = 0.0468), Teye and nose length after run (p = 0.0076), and Tmouth and nose length after run (p = 0.0110). As body temperature rises during exercise, it becomes increasingly important to regulate blood flow throughout the body to supply working tissues with oxygen. This demand is offset by the role of the snout in evaporative cooling through panting, functioning as a prime location for heat dissipation and therefore maintaining significant relationships with many other vascularized tissues.

A revisiting of "the hallmarks of aging" in domestic dogs: current status of the literature.

A progressive decline in biological function and fitness is, generally, how aging is defined. However, in 2013, a description on the "hallmarks of aging" in mammals was published, and within it, it described biological processes that are known to alter the aging phenotype. These include genomic instability, telomere attrition, epigenetic alterations, loss of proteostasis, deregulated nutrient sensing, mitochondrial dysfunction, cellular senescence, stem cell exhaustion, altered intercellular communication (inflammation), and changes within the microbiome. This mini-review provides a detailed account of the progress on each of these hallmarks of aging in the domestic dog within the last 5 years. Additionally, when there are gaps in the literature between other mammalian species and dogs, I highlight the aging biomarkers that may be missing for dogs as aging models. I also argue for the importance of dog aging studies to include several breeds of dogs at differing ages and for age corrections for breeds with differing mean lifespans throughout.

Cellular metabolic pathways of aging in dogs: could p53 and SIRT1 be at play?

Aging and cancer seem to be closely associated, such that cancer is generally considered a disease of the elderly in both humans and dogs. Additionally, cancer is a metabolic shift in itself towards aerobic glycolysis. Larger dog breeds with shorter lifespans, and increased glycolytic cellular metabolic rates, die of cancer more often than smaller breeds. The tumor suppressor p53 factor is a key suppressor oncogene, and the p53 pathway arrests cellular proliferation and prevents DNA mutations from accumulating during cellular stress. The p53 pathway is also associated with the control of cellular metabolism to prevent cellular metabolic shifts common to cancerous phenotypes. SIRT1 deacetylates the p53 tumor suppressor protein, downregulating p53 via effects on stability and activity during stress. Here, we used primary fibroblast cells from small and large puppies and old dogs. Using UV radiation to upregulate the p53 system (100 J/m2), control cells and UV-treated cells were used to measure aerobic and glycolytic metabolic rates using a Seahorse XFe96 oxygen flux analyzer. We also quantified p53 expression and SIRT1 concentration in canine primary fibroblasts before and after UV treatment. We demonstrate that, due to a higher p53 nuclear to cytoplasmic ratio in large breed dogs after UV treatment, p53 could have a more regulatory effect on large breed dogs' metabolism compared with smaller breeds. Thus, there may be a link between p53 upregulation and inhibition of glycolysis in large breed dogs during times of cellular stress compared with small breed dogs. However, SIRT1 concentrations decrease with age in domestic dogs of both size classes, suggesting a possible release of inhibition of p53 through the SIRT1 pathway with age. This may lead to increased incidences of cancer, especially due to the more pronounced upregulation of p53 with cellular stress.

Chronic Thermal Acclimation Effects on Critical Thermal Maxima (CTmax) and Oxidative Stress Differences in White Epaxial Muscle between Surface and Cave Morphotypes of the Mexican Cavefish (Astyanax mexicanus).

AbstractIn the face of increasing environmental temperatures, operative differences between mitochondrial function and whole-animal phenotypic response to the environment are underrepresented in research, especially in subtemperate ectothermic vertebrates. A novel approach to exploring this connection is to examine model species that are genetically similar but that have different whole-animal phenotypes, each of which inhabits different environments. The blind Mexican cavefish (Astyanax mexicanus) has the following two morphotypes: a surface form found in aboveground rivers and an obligate cave-dwelling form. Each morphotype inhabits vastly different thermal and oxygen environments. Whole-animal and mitochondrial responses to thermal acclimation and oxidative stress, with respect to increasing temperatures, have not been previously determined in either morphotype of this species. Here, we chronically acclimated both morphotypes to three temperatures (14°C, 25°C, and 31°C) to establish potential for acclimation and critical thermal maxima (CTmax) for each morphotype of this species. After measuring CTmax in six cohorts, we additionally measured enzymatic antioxidant capacity (catalase, superoxide dismutase, and glutathione peroxidase activities), peroxyl scavenging capacity, and lipid peroxidation damage in white epaxial muscle for each individual. We found a significant effect of acclimation temperature on CTmax (F=29.57, P<0.001) but no effect of morphotype on CTmax (F=2.092, P=0.162). Additionally, we found that morphotype had a significant effect on glutathione peroxidase activity, with the surface morphotype having increased glutathione peroxidase activity compared with the cave morphotype (F=6.270, P=0.020). No other oxidative stress variable demonstrated significant differences. Increases in CTmax with chronic thermal acclimation to higher temperatures suggests that there is some degree of phenotypic plasticity in this species that nominally occupies thermally stable environments. The decreased glutathione peroxidase activity in the cave morphotype may be related to decreased environmental oxygen concentration and decreased metabolic rate in this environmentally constrained morphotype compared to in its surface-living counterparts.

White epaxial muscle aerobic and anaerobic potential and muscle fiber structure in surface and cave morphotypes of the Mexican cavefish (Astyanax mexicanus).

Proper muscle function and muscle fiber structures that match the environmental demands of organisms are imperative to their success in any ecosystem. The Mexican cavefish, Astyanax mexicanus, has two morphotypes: an obligate cave-dwelling form that lives in thermally insulated caves and an O2 poor environment, and a surface form that lives in a more thermally variable, but O2 rich river environment. As environment can determine physiological adaptations, it is of interest to compare the aerobic and anaerobic metabolic profiles of white muscle metabolism in both morphotypes of this species, as well as their muscle structures. Here, we used white muscle of both morphotypes of the Mexican cavefish to determine citrate synthase (CS) activity as a measure of aerobic potential, and lactate concentration as a measure of anaerobic potential at three different chronic acclimation temperatures (14°C, 25°C, and 31°C). By examining aerobic and anaerobic potential in both morphs, we sought to link environmental thermal flexibility to muscle metabolism. We found that the surface morphotype had higher CS activity and lower lactate concentration, suggesting an overall more efficient usage of aerobic metabolism; whereas the cave morphotype showed lower CS activity and higher lactate concentration, suggesting a stronger reliance on anaerobic pathways. We also measured white muscle histological variables that have been previously linked to whole-animal metabolism: fiber diameter, number of nuclei per mm of fiber and myonuclear domain (MND) of both morphotypes at 25°C to examine cell-level differences in muscle morphology. However, we found no differences in fiber diameter, number of nuclei per mm of fiber or MND between the two morphotypes. Thus, although the cellular morphology is similar in these species, the environmental differences in the evolution of the two morphs has led to differences in their metabolic profiles.

Enzymatic responses reveal different physiological strategies employed by eurytolerant fish during extreme hot and cold cycling acclimation temperatures.

Heat waves and cold snaps are projected to rise in magnitude, duration, interval, and harshness in the coming years. The current literature examining thermal impacts on the physiology of organisms rarely uses chronic, variable thermal acclimations despite the fact that climate change predictions project a more variable environment. If we are to determine species' susceptibility to climate change, chronic and variable lab acclimations should be prioritized. Here, we acclimated the eurytolerant sheepshead minnow (Cyprinodon variegatus) to two extreme cycling thermal regimes: one warm [resting 27 °C with a spike to 33 °C for 8 h daily], one cold [resting 6.5 °C with a spike to 12 °C for 8 h daily], and three chronically stable conditions (10, 22, and 30 °C) for comparison. We measured enzymatic antioxidants (catalase (CAT), superoxide dismutase (SOD) and glutathione peroxidase (GPx)), total antioxidant capacity, lipid peroxidation (LPO) damage, and citrate synthase (CS) activity in white epaxial muscle. Of particular note, we found significant increases in log CAT activity and SOD concentration in the warm cycling temperatures, and significant increases in GPx activity in the cold cycling temperatures. We found no significant accumulation of LPO damage in any of our thermal acclimation treatments. Thus, sheepshead minnows demonstrate two particularly different mechanisms towards dealing with thermal variation in low and high temperatures. The enzymatic differences between low and high cycling temperatures may define pathways of eurytolerant organisms and how they may survive predicted variability in thermal regimes.

Inflammaging in domestic dogs: basal level concentrations of IL-6, IL-1ß, and TNF-α in serum of healthy dogs of different body sizes and ages.

Inflammaging, a "hallmark of aging," refers to a chronic, progressive increase in the proinflammatory status of mammals as they age, and this phenotype has been associated with many age-related diseases, including cardiovascular disease, arthritis and cancer. Though, inflammaging research is common in humans, there is a lack of data for this process for the domestic dog. Here, serum concentrations of IL-6, IL-1ß and TNF-α in healthy dogs of different body sizes and ages were measured to determine whether inflammaging may play a mechanistic role in aging rates in dogs, similar to those found in humans. Using a four-way ANOVA, a significant decrease in IL-6 concentrations in young dogs with the rest of the age categories showing increased IL-6 concentrations was found, similar to humans. However, only young dogs have decreased IL-6 concentrations, with adult dogs having similar IL-6 concentrations to senior and geriatric dogs, implying differences in aging rates between humans and dogs. And, there was a marginally significant interaction between sex*spayed or neutered status and IL-1ß concentrations with intact females having the lowest IL-1ß concentrations compared with intact males, and spayed and neutered dogs. The presence of estrogen in intact females may, overall, decrease inflammatory pathways. This implies that age at spaying or neutering may be an important aspect to consider for inflammaging pathways in dogs. Furthermore, sterilized dogs often die of immune-related diseases, which could be linked to the increases in IL-1ß in sterilized dogs found in this study.

Exploring the role of primary fibroblast cells in comparative physiology: a historical and contemporary overview.

With the advent of tissue culture, and eventually the in vitro growth and maintenance of individual cell types, it became possible to ask mechanistic questions about whole organism physiology that are impractical to address within a captive setting or within the whole organism. The earliest studies focused on understanding the wound-healing response while refining cell growth and maintenance protocols from various species. In addition to its extensive use in biomedical research, this approach has been co-opted by comparative physiologists interested in reductionist/mechanistic questions related to how cellular physiology can help explain whole organism function. Here, we provide a historical perspective on the emergence of primary cell culture with an emphasis on fibroblasts followed by an overview of applying this method to ask questions about the role of life-history evolution in shaping organismal physiology at the cellular level, as well as the effect of exogenous factors (i.e., temperature, and oxygen availability) on cellular function. Finally, we propose future uses for primary fibroblasts to address questions in conservation biology and comparative physiology.

Can dogs serve as stress mediators to decrease salivary cortisol levels in a population of liberal arts college undergraduate students?

The steroid hormone cortisol can be used to measure physiological stress in humans. The hypothalamic-pituitary-adrenal (HPA) axis synthesizes cortisol, and a negative feedback cycle regulates cortisol depending on an individual's stress level and/or circadian rhythm. Chronic stress of college undergraduate students is associated with various adverse health effects, including anxiety and depression. Reports suggest that stress levels have risen dramatically in recent years, particularly among university students dealing with intense academic loads in addition to COVID-19 pandemic-related uncertainty. The increasing rate of mental illness on college campuses necessitates the study of mediators potentially capable of lowering stress, and thus cortisol levels. Research on mediation techniques and coping mechanisms have gained traction to address the concerning levels of stress, including the employment of human-animal interaction sessions on college campuses. In this study, human-canine interaction as a stress mediation strategy for undergraduate students was investigated. We measured salivary cortisol levels in 73 college undergraduate students during a 60-min interaction period with a dog to determine whether human-canine interactions are effective in lowering cortisol levels and potentially reducing chronic stress typical of undergraduate students. Our results indicate that a human-canine interaction for 60 min is an effective method for significantly reducing salivary cortisol and stress levels among undergraduate college students. These findings support the expansion of animal visitation programs on college campuses to help students manage stress.

Scaling with body mass and age in glycolytic enzymes of domestic dogs.

Animals produce ATP through oxidative phosphorylation using oxygen, but cellular energy can also be obtained through glycolysis when oxygen is not present at sufficient levels. Although most mammals of larger body mass have longer life spans, small dog breeds tend to outlive large breeds. Primary fibroblast cells from larger breeds of dogs have previously been shown to have increased dependency on glycolytic phenotypes across their lifespan. Different levels of activity of the glycolytic enzymes pyruvate kinase (PK), lactate dehydrogenase (LDH), and phosphoenolpyruvate carboxykinase (PEPCK) may provide insight to a mechanism that leads to the different metabolic phenotype observed in different sized breeds as they age. In this study, 1) we measured the activities of PK, LDH, and PEPCK in primary fibroblasts from dogs of different breed sizes and age classes and 2) measured the activities of PK and LDH in plasma from dogs of different breed sizes and age classes. We found that there was no significant relationship between body mass and PK, LDH and PEPCK activity in primary fibroblasts. Further, there were not significant differences with activity in these enzymes for old dogs compared to young dogs. In plasma, we found a negative correlation between PK activity and body mass and no relationship between LDH activity and body mass. There was a negative relationship between LDH activity and age in dogs. Further, while a negative correlational relationship between PK activity and age was only marginal, a best subsets regression model demonstrated a significant marginal effect of age on PK activity. PK and LDH may provide intermediates for other metabolic pathways in small breeds. However, large breed dogs may demonstrate a deficiency in metabolism at the PK level, a cellular metabolic pathway that may potentially aid in tumor progression.

Effect of different masses, ages, and coats on the thermoregulation of dogs before and after exercise across different seasons.

Domestic dogs demonstrate a positive correlation between body mass and whole-animal metabolism but a negative relationship between body mass and lifespan in dogs. Additionally, essential physiological mechanisms in domestic dogs, such as the relationships between thermal relations and body size and age, remain poorly understood. In this study, we looked at thermoregulation in dogs of different sizes, ages, coat types, and head morphologies across three different seasons. We used tympanic membrane temperatures (Tear) and infrared thermography to observe temperature regulation in pet dogs before and after a 45-min moderate walking exercise trial. We hypothesized that Tear and heat dissipation is positively correlated with body mass. Using network analysis, we found that body mass was among the most central features for spring and summer trials, but not for the winter trials. Similarly, leg length, snout length, and paw width were the central predictors in two of the three seasons. Mediation analysis demonstrated that nose and snout length act as significant mediators of the effects of body mass on mouth temperatures in the spring. For the summer trials, nose length and paw width significantly mediated the effect of body mass on mouth temperatures. Age, however, does not seem to be a major determinant of thermoregulation in dogs according to best subset models. A cross-seasonal examination of repeated measurements showed that mouth temperature heat dissipation rates decreased with increasing temperature and humidity. Overall, our findings support our hypothesis that Tear and heat dissipation rates are positively correlated with body mass in dogs, thus, negatively correlated with mass-specific metabolism. This finding suggests that small dogs allocate a bigger proportion of their metabolism to "inefficiencies" of heat production to offset greater heat loss.

Skeletal muscle and metabolic flexibility in response to changing energy demands in wild birds.

Phenotypically plastic responses of animals to adjust to environmental variation are pervasive. Reversible plasticity (i.e., phenotypic flexibility), where adult phenotypes can be reversibly altered according to prevailing environmental conditions, allow for better matching of phenotypes to the environment and can generate fitness benefits but may also be associated with costs that trade-off with capacity for flexibility. Here, we review the literature on avian metabolic and muscle plasticity in response to season, temperature, migration and experimental manipulation of flight costs, and employ an integrative approach to explore the phenotypic flexibility of metabolic rates and skeletal muscle in wild birds. Basal (minimum maintenance metabolic rate) and summit (maximum cold-induced metabolic rate) metabolic rates are flexible traits in birds, typically increasing with increasing energy demands. Because skeletal muscles are important for energy use at the organismal level, especially to maximum rates of energy use during exercise or shivering thermogenesis, we consider flexibility of skeletal muscle at the tissue and ultrastructural levels in response to variations in the thermal environment and in workloads due to flight exercise. We also examine two major muscle remodeling regulatory pathways: myostatin and insulin-like growth factor -1 (IGF-1). Changes in myostatin and IGF-1 pathways are sometimes, but not always, regulated in a manner consistent with metabolic rate and muscle mass flexibility in response to changing energy demands in wild birds, but few studies have examined such variation so additional study is needed to fully understand roles for these pathways in regulating metabolic flexibility in birds. Muscle ultrastrutural variation in terms of muscle fiber diameter and associated myonuclear domain (MND) in birds is plastic and highly responsive to thermal variation and increases in workload, however, only a few studies have examined ultrastructural flexibility in avian muscle. Additionally, the relationship between myostatin, IGF-1, and satellite cell (SC) proliferation as it relates to avian muscle flexibility has not been addressed in birds and represents a promising avenue for future study.

Differences in advanced glycation endproducts (AGEs) in plasma from birds and mammals of different body sizes and ages.

Birds and mammals provide a physiological paradox: similar-sized mammals live shorter lives than birds; yet, birds have higher blood glucose concentrations than mammals, and higher basal metabolic rates. We have previously shown that oxidative stress patterns between mammals and birds differ, so that birds, generally, have lower blood antioxidant capacity, and lower lipid peroxidation concentration. There is a close association between oxidative stress and the production of carbohydrate-based damaged biomolecules, Advanced Glycation End-products (AGEs). In mammals, AGEs can bind to their receptor (RAGE), which can lead to increases in reactive oxygen species (ROS) production, and can decrease antioxidant capacity. Here, we used plasma from birds and mammals to address whether blood plasma AGE-BSA concentration is associated with body mass and age in these two groups. We found a statistically significantly higher average concentrations of AGE-BSA in birds compared with mammals, and we found a significantly positive correlation between AGE-BSA and age in mammals, though, this correlation disappeared after phylogenetic correction. We propose that the higher AGE concentration in birds is mainly attributable to greater AGE-production due to elevated basal glucose concentrations and decreased AGE-clearance given differences in glomerular filtration rates in birds compared with mammals. Additionally, due to the potential lack of an AGE receptor in birds, AGE accumulation may not be closely linked to oxidative stress and therefore pose a lesser physiological challenge in birds compared to mammals.

Lack of variation in nuclear DNA content in avian muscle.

The avian pectoralis muscle demonstrates plasticity with regard to size, so that temperate birds facing winter conditions or birds enduring a migration bout tend to have significant increases in the size and mass of this tissue due to muscular hypertrophy. Myonuclear domain (MND), the volume of cytoplasm a myonuclei services, in the pectoralis muscle of birds seems to be altered during thermal stress or changing seasons. However, there is no information available regarding muscle DNA content or ploidy level within the avian pectoralis. Changes in muscle DNA content can be used in this tissue to aid in size and mass changes. Here, we hypothesized that long-distance migrants or temperate residents would use the process of endoreduplication to aid in altering muscle size. Mostly contradictory to our hypotheses, we found no differences in the mean muscle DNA content in any of the 62 species of birds examined in this study. We also found no correlations between mean muscle DNA content and other muscle structural measurements, such as the number of nuclei per millimeter of fiber, myonuclear domain, and fiber cross-sectional area. Thus, while avian muscle seems more phenotypically plastic than mammalian muscle, the biological processes surrounding myonuclear function may be more closely related to those seen in mammals.

Altered Oxidative Status as a Cost of Reproduction in a Seabird with High Reproductive Costs.

AbstractLife history theory posits that reproduction is constrained by a cost of reproduction such that any increase in breeding effort should reduce subsequent survival. Oxidative stress refers to an imbalance between the prooxidant reactive oxygen species (ROS) and antioxidant defense. If not thwarted, ROS can cause damage to DNA, lipids, and proteins, potentially increasing the rate of senescence and decreasing cellular function. Reproduction is often associated with higher metabolic rates, which could increase production of ROS and lead to oxidative damage if the animal does not increase antioxidant protection. Thus, oxidative stress could be one mechanism creating a cost of reproduction. In this study we explored how reproduction may affect oxidative status differently between male and female thick-billed murres during early and late breeding seasons over three consecutive years. We manipulated breeding efforts by removing an egg from the nest of some individuals, which forced females to relay, and by handicapping other individuals by clipping wings. We measured total antioxidant capacity (TAC), uric acid (UA) concentration, and malondialdehyde (MDA; an index of lipid oxidative damage) concentration in blood plasma as well as activities of superoxide dismutase (SOD), glutathione peroxidase (GPx), and catalase (CAT) in red blood cells. Oxidative status was highly variable across years, and year was consistently the most important factor determining oxidative status; inconsistent results in previous field studies may be because reproductive oxidative stress occurs only in some years. Females had lower SOD and GPx and higher MDA and TAC than males immediately after egg laying, suggesting that the cost of egg laying required investment in cheaper nonenzymatic antioxidant defenses that had lower capacity for defending against lipid peroxidation. Delayed birds had lower UA and lower SOD, GPx, and CAT activity compared with control birds. In conclusion, when reproductive costs increase via higher energy costs or longer breeding seasons, the oxidative status of both male and female murres deteriorated as a result of reduced antioxidant defenses.

Effects of acute temperature increases on House sparrow (Passer domesticus) pectoralis muscle myonuclear domain.

With rapid climate change, heat wave episodes have become more intense and more frequent. This poses a significant threat to animals, and forces them to manage these physiologically challenging conditions by adapting and/or moving. As an invasive species with a large niche breadth, House sparrows (Passer domesticus) exhibit high phenotypic flexibility that caters to seasonal changes in function and metabolism. For example, their pectoral muscle complex exhibits size and mass plasticity with winter and summer acclimation. Here, we investigated the effects of acute whole-organism heat stress to 43°C on cellular-level changes in House sparrow pectoralis muscle myonuclear domain (MND), the volumetric portion each nucleus is responsible for, that have gone overlooked in the current literature. House sparrows were separated into a control group, a heat-shocked group subjected to thermal stress at 43°C for 24 h, and a recovery group that was returned to room temperature for 24 h after experiencing the same temperature treatment. Here, we found that heat-shocked and recovery groups demonstrated a decrease in number of nuclei per millimeter of fiber and increase in MND, when compared with the control. We also found a significant positive correlation between fiber diameter and MND in the recovery group, suggesting the possibility that nuclei number constrains the extent of muscle fiber size. Together, these results show that acute heat shock alters House sparrow pectoralis muscle cellular physiology in a rigid way that could prove detrimental to long-term muscle integrity and performance.

Do thermal acclimation and an acute heat challenge alter myonuclear domain of control- and fast-growing quail?

Efforts to determine physiological traits that may render species resilient or susceptible to changing global temperatures have accelerated in recent years. Temperature is of critical importance to biological function; thus, climate change has the potential to severely affect all levels of biological organization in many species. For example, increases in environmental temperatures may alter muscle structure and function in birds. Myonuclear domain (MND), an under-studied aspect of avian muscle physiology that changes in response to thermal stress, is defined as the amount of cytoplasm within a muscle fiber that each nucleus is responsible for servicing. Here, we used two random bred lines of Japanese quail (Coturnix japonica) representing examples of control and fast growth rates. We used a factorial design to administer four treatment combinations to each line - an initial period of either heat-stress acclimation (Acclimation) or no acclimation (Not acclimated) followed by either a heat-stress challenge (HS) or no challenge (NC) after week 8 of age - to determine the effects of thermal acclimation and acute thermal stress on quail MND. We found a significant interaction between line * final treatment with fast-growing, HS birds demonstrating the lowest numbers of nuclei per mm of fiber, and Acclimated control-growing birds showing the highest numbers of nuclei per mm of fiber. There was a significant effect of line on MND with the fast-growing line having larger MNDs. Initial treatment with Not Acclimated birds showed larger MNDs. Additionally, control growing quail demonstrated positive correlations with fiber size, whereas fast growing quail did not. This may mean that nuclei in larger fibers of fast-growing quail may be functioning maximally, and that increases in temperature may also demonstrate similar effects.

Metabolomics of aging in primary fibroblasts from small and large breed dogs.

Among several animal groups (eutherian mammals, birds, reptiles), lifespan positively correlates with body mass over several orders of magnitude. Contradicting this pattern are domesticated dogs, with small dog breeds exhibiting significantly longer lifespans than large dog breeds. The underlying mechanisms of differing aging rates across body masses are unclear, but it is generally agreed that metabolism is a significant regulator of the aging process. Herein, we performed a targeted metabolomics analysis on primary fibroblasts isolated from small and large breed young and old dogs. Regardless of size, older dogs exhibited lower glutathione and ATP, consistent with a role for oxidative stress and bioenergetic decline in aging. Furthermore, several size-specific metabolic patterns were observed with aging, including the following: (i) An apparent defect in the lower half of glycolysis in large old dogs at the level of pyruvate kinase. (ii) Increased glutamine anaplerosis into the TCA cycle in large old dogs. (iii) A potential defect in coenzyme A biosynthesis in large old dogs. (iv) Low nucleotide levels in small young dogs that corrected with age. (v) An age-dependent increase in carnitine in small dogs that was absent in large dogs. Overall, these data support the hypothesis that alterations in metabolism may underlie the different lifespans of small vs. large breed dogs, and further work in this area may afford potential therapeutic strategies to improve the lifespan of large dogs.

Effects of metformin, rapamycin, and resveratrol on cellular metabolism of canine primary fibroblast cells isolated from large and small breeds as they age.

Small breed dogs have longer lifespans than their large breed counterparts. Previous work demonstrated that primary fibroblast cells isolated from large breed young and old dogs have a persistent glycolytic metabolic profile compared with cells from small breed dogs. Here, we cultured primary fibroblast cells from small and large, young and old dogs and treated these cells with three commercially available drugs that show lifespan and health span benefits, and have been shown to reduce glycolytic rates: rapamycin (rapa), resveratrol (res) and metformin (met). We then measured aerobic and anaerobic cellular respiration in these cells. We found that rapa and res increased rates of non-glycolytic acidification in small and large breed puppies and basal oxygen consumption rates (OCR) in small and large breed puppies. Rapa increased proton leak and non-mitochondrial respiration in small and large breed puppies. Maximal respiration was significantly altered with rapa treatment but in opposing ways: large breed puppies showed a significant increase in maximal respiration when treated with rapa, and small old dogs demonstrated a significant decrease in maximal respiration when treated with rapa. In opposition to rapa treatments, met significantly decreased basal OCR levels in cells from small and large breed puppies. Our data suggest that rapa treatments may be metabolically beneficial to dogs when started early in life and more beneficial in larger breeds.

The Physiological Conundrum That is the Domestic Dog.

Across Mammalia, body size and lifespan are positively correlated. However, in domestic dogs, the opposite is true: small dogs have longer lives compared with large dogs. Here, I present literature-based data on life-history traits that may affect dog lifespan, including adaptations at the whole-organism, and organ-level. Then, I compare those same traits to wild canids. Because oxidative stress is a byproduct of aerobic metabolism, I also present data on oxidative stress in dogs that suggests that small breed dogs accumulate significantly more circulating lipid peroxidation damage compared with large breed dogs, in opposition to lifespan predictions. Further, wild canids have increased antioxidant concentrations compared with domestic dogs, which may aid in explaining why wild canids have longer lifespans than similar-sized domestic dogs. At the cellular level, I describe mechanisms that differ across size classes of dogs, including increases in aerobic metabolism with age, and increases in glycolytic metabolic rates in large breed dogs across their lifespan. To address potential interventions to extend lifespan in domestic dogs, I describe experimental alterations to cellular architecture to test the "membrane pacemaker" hypotheses of metabolism and aging. This hypothesis suggests that increased lipid unsaturation and polyunsaturated fatty acids in cell membranes can increase cellular metabolic rates and oxidative damage, leading to potential decreased longevity. I also discuss cellular metabolic changes of primary fibroblast cells isolated from domestic dogs as they are treated with commercially available drugs that are linked to lifespan and health span expansion.