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.

Dietary vitamin E reaches the mitochondria in the flight muscle of zebra finches but only if they exercise.

Whether dietary antioxidants are effective for alleviating oxidative costs associated with energy-demanding life events first requires they are successfully absorbed in the digestive tract and transported to sites associated with reactive species production (e.g. the mitochondria). Flying birds are under high energy and oxidative demands, and although birds commonly ingest dietary antioxidants in the wild, the bioavailability of these consumed antioxidants is poorly understood. We show for the first time that an ingested lipophilic antioxidant, α-tocopherol, reached the mitochondria in the flight muscles of a songbird but only if they regularly exercise (60 min of perch-to-perch flights two times in a day or 8.5 km day-1). Deuterated α-tocopherol was found in the blood of exercise-trained zebra finches within 6.5 hrs and in isolated mitochondria from pectoral muscle within 22.5 hrs, but never reached the mitochondria in caged sedentary control birds. This rapid pace (within a day) and extent of metabolic routing of a dietary antioxidant to muscle mitochondria means that daily consumption of such dietary sources can help to pay the inevitable oxidative costs of flight muscle metabolism, but only when combined with regular exercise.

Acute effects of intense exercise on the antioxidant system in birds: does exercise training help?

The acute effects of an energy-intensive activity such as exercise may alter an animal's redox homeostasis, although these short-term effects may be ameliorated by chronic exposure to that activity, or training, over time. Although well documented in mammals, how energy-intensive training affects the antioxidant system and damage by reactive species has not been investigated fully in flight-trained birds. We examined changes to redox homeostasis in zebra finches exposed to energy-intensive activity (60 min of perch-to-perch flights twice a day), and how exercise training over many weeks affected this response. We measured multiple components of the antioxidant system: an enzymatic antioxidant (glutathione peroxidase, GPx) and non-enzymatic antioxidants (measured by the OXY-adsorbent test) as well as a measure of oxidative damage (d-ROMs). At no point during the experiment did oxidative damage change. We discovered that exposure to energy-intensive exercise training did not alter baseline levels of GPx, but induced exercise-trained birds to maintain a higher non-enzymatic antioxidant status as compared with untrained birds. GPx activity was elevated above baseline in trained birds immediately after completion of the second 1 h flight on each of the three sampling days, and non-enzymatic antioxidants were acutely depleted during flight after 13 and 44 days of training. The primary effect of exercise training on the acute response of the antioxidant system to 2 h flights was increased coordination between the enzymatic (GPx) and non-enzymatic components of the antioxidant system of birds that reduced oxidative damage associated with exercise.

Dynamics of Individual Fatty Acids in Muscle Fat Stores and Membranes of a Songbird and Its Functional and Ecological Importance.

Although tissue fatty acid (FA) composition has been linked to whole-animal performance (e.g., aerobic endurance, metabolic rate, postexercise recovery) in a wide range of animal taxa, we do not adequately understand the pace of changes in FA composition and its implications for the ecology of animals. Therefore, we used a C4 to C3 diet shift experiment and compound-specific δ13C analysis to estimate the turnover rates of FAs in the polar and neutral fractions of flight muscle lipids (corresponding to membranes and lipid droplets) of exercised and sedentary zebra finches (Taeniopygia guttata). Turnover was fastest for linoleic acid (LA; 18:2n6) and palmitic acid (PA; 16:0), with 95% replacement times of 10.8-17.7 d in the polar fraction and 17.2-32.8 d in the neutral fraction, but was unexpectedly slow for the long-chain polyunsaturated FAs (LC-PUFAs) arachidonic acid (20:4n6) and docosahexaenoic acid (22:6n3) in the polar fraction, with 95% replacement in 64.9-136.5 d. Polar fraction LA and PA turnover was significantly faster in exercised birds (95% replacement in 8.5-13.3 d). Our results suggest that FA turnover in intramuscular lipid droplets is related to FA tissue concentrations and that turnover does not change in response to exercise. In contrast, we found that muscle membrane FA turnover is likely driven by a combination of selective LC-PUFA retention and consumption of shorter-chain FAs in energy metabolism. The unexpectedly fast turnover of membrane-associated FAs in muscle suggests that songbirds during migration could substantially remodel their membranes within a single migration stopover, and this may have substantial implications for how the FA composition of diet affects energy metabolism of birds during migration.

Turnover of muscle lipids and response to exercise differ between neutral and polar fractions in a model songbird, the zebra finch.

The turnover rates of tissues and their constituent molecules give us insights into animals' physiological demands and their functional flexibility over time. Thus far, most studies of this kind have focused on protein turnover, and few have considered lipid turnover despite an increasing appreciation of the functional diversity of this class of molecules. We measured the turnover rates of neutral and polar lipids from the pectoralis muscles of a model songbird, the zebra finch (Taeniopygia guttata, N=65), in a 256 day C3/C4 diet shift experiment, with tissue samples taken at 10 time points. We also manipulated the physiological state of a subset of these birds with a 10 week flight training regimen to test the effect of exercise on lipid turnover. We measured lipid δ13C values via isotope ratio mass spectrometry (IRMS) and estimated turnover in different fractions and treatment groups with non-linear mixed-effect regression. We found a significant difference between the mean retention times (τ) of neutral and polar lipids (t119=-2.22, P=0.028), with polar lipids (τ=11.80±1.28 days) having shorter retention times than neutral lipids (τ=19.47±3.22 days). When all birds were considered, we also found a significant decrease in the mean retention time of polar lipids in exercised birds relative to control birds (difference=-2.2±1.83 days, t56=-2.37, P=0.021), but not neutral lipids (difference=4.2± 7.41 days, t56=0.57, P=0.57). A larger, more variable neutral lipid pool and the exposure of polar lipids in mitochondrial membranes to oxidative damage and increased turnover provide mechanisms consistent with our results.

The role of the antioxidant system during intense endurance exercise: lessons from migrating birds.

During migration, birds substantially increase their metabolic rate and burn fats as fuel and yet somehow avoid succumbing to overwhelming oxidative damage. The physiological means by which vertebrates such as migrating birds can counteract an increased production of reactive species (RS) are rather limited: they can upregulate their endogenous antioxidant system and/or consume dietary antioxidants (prophylactically or therapeutically). Thus, birds can alter different components of their antioxidant system to respond to the demands of long-duration flights, but much remains to be discovered about the complexities of RS production and antioxidant protection throughout migration. Here, we use bird migration as an example to discuss how RS are produced during endurance exercise and how the complex antioxidant system can protect against cellular damage caused by RS. Understanding how a bird's antioxidant system responds during migration can lend insights into how antioxidants protect birds during other life-history stages when metabolic rate may be high, and how antioxidants protect other vertebrates from oxidative damage during endurance exercise.

The metabolic rate of cultured muscle cells from hybrid Coturnix quail is intermediate to that of muscle cells from fast-growing and slow-growing Coturnix quail.

Growth rate is a fundamental parameter of an organism's life history and varies 30-fold across bird species. To explore how whole-organism growth rate and the metabolic rate of cultured muscle cells are connected, two lines of Japanese quail (Coturnix coturnix japonica), one that had been artificially selected for fast growth for over 60 generations and a control line were used to culture myoblasts. In line with previous work, myoblasts from the fast growth line had significantly higher rates of oxygen consumption, glycolytic flux, and higher mitochondrial volume than myoblasts from the control line, indicating that an increase in growth rate is associated with a concomitant increase in cellular metabolic rates and that mitochondrial density contributes to the differences in rates of metabolism between the lines. We reared chicks from two hybrid lines with reciprocal parental configurations for growth rate to explore the effect of maternally inherited mitochondrial DNA on rates of growth and metabolism. Growth rates of chicks, cellular basal oxygen consumption, glycolytic flux, and mitochondrial volume in myoblasts from chicks from both reciprocal crosses were intermediate to the fast and control lines. This indicates that genes in the nucleus have a strong influence on metabolic rates at the cellular level, compared with maternally inherited mitochondrial DNA.

Physiological underpinnings associated with differences in pace of life and metabolic rate in north temperate and neotropical birds.

Animal life-history traits fall within limited ecological space with animals that have high reproductive rates having short lives, a continuum referred to as a "slow-fast" life-history axis. Animals of the same body mass at the slow end of the life-history continuum are characterized by low annual reproductive output and low mortality rate, such as is found in many tropical birds, whereas at the fast end, rates of reproduction and mortality are high, as in temperate birds. These differences in life-history traits are thought to result from trade-offs between investment in reproduction or self-maintenance as mediated by the biotic and abiotic environment. Thus, tropical and temperate birds provide a unique system to examine physiological consequences of life-history trade-offs at opposing ends of the "pace of life" spectrum. We have explored the implications of these trade-offs at several levels of physiological organization including whole-animal, organ systems, and cells. Tropical birds tend to have higher survival, slower growth, lower rates of whole-animal basal metabolic rate and peak metabolic rate, and smaller metabolically active organs compared with temperate birds. At the cellular level, primary dermal fibroblasts from tropical birds tend to have lower cellular metabolic rates and appear to be more resistant to oxidative cell stress than those of temperate birds. However, at the subcellular level, lipid peroxidation rates, a measure of the ability of lipid molecules within the cell membranes to thwart the propagation of oxidative damage, appear not to be different between tropical and temperate species. Nevertheless, lipids in mitochondrial membranes of tropical birds tend to have increased concentrations of plasmalogens (phospholipids with antioxidant properties), and decreased concentrations of cardiolipin (a complex phospholipid in the electron transport chain) compared with temperate birds.

Cellular metabolic rates in cultured primary dermal fibroblasts and myoblast cells from fast-growing and control Coturnix quail.

Fibroblast cells have been extensively used in research, including in medicine, physiology, physiological-ecology, and conservation biology. However, whether the physiology of fibroblasts reflects the physiology of other cell types in the same animal is unknown. Dermal fibroblasts are responsible for generating connective tissue and involved in wound healing, but generally, this cell type is thought to be metabolically inactive until it is required at the site of tissue damage. Thus, one might question whether fibroblasts are a representative model system to portray the metabolic profile of the whole organism, as compared with cells isolated from other tissues, like muscle, brain or kidneys. To explore whether fibroblasts have the same metabolic profile as do myoblast cells, we cultured cells from day-old chicks of quail (Coturnix coturnix japonica) selected for fast-growth or normal growth (our control group). Our results suggest that isolated primary fibroblasts and myoblast cells had higher rates of glycolysis, oxygen consumption and more mitochondria in the fast-growing line than in the control line. Our findings lend support for the idea that fibroblasts are a representative cell system to characterize the whole organism metabolic signature at the cellular-level. These data are striking, however, because fibroblasts had higher rates of metabolism for every parameter measured than myoblast cells isolated from the same individuals.