High-altitude Mice Research Articles

Metabolic recovery from submaximal exercise in hypoxia acclimated high altitude deer mice (Peromyscus maniculatus)

Animals living at high-altitude are faced with unremitting low oxygen availability. This can make it difficult to perform daily tasks that require increases in aerobic metabolism. An activity important for survival is aerobic locomotion, and the rapid recovery of muscle metabolism post exercise. Past work shows that hypoxia acclimated high-altitude mice (Peromyscus maniculatus) have a greater reliance on carbohydrates to power exercise than low altitude mice. However, it is unclear how quickly after aerobic exercise these mice can recovery and replenish muscle glycogen stores. The gastrocnemius muscle of high-altitude deer mice has a more aerobic phenotype and a greater capacity to oxidize lipids than low altitude deer mice. This suggests that high altitude mice may recover more rapidly from exercise than their lowland counterparts due to a greater capacity to support glycogen replenishment using intramuscular triglycerides (IMTG). To explore this possibility, we used low- and high-altitude native deer mice born and raised in common lab conditions and acclimated to chronic hypoxia. We determined changes in oxygen consumption following 15 min of aerobic exercise in 12% O2 and sampled skeletal muscles and liver at various time points during recovery to examine changes in key metabolites, including glycogen and IMTG. We found depletion in glycogen stores during exercise only in lowlanders, which returned to resting levels following 90 min of recovery. In contrast, IMTG did not change significantly with exercise or during recovery in either population. These data suggest that exercise recovery is influenced by altitude ancestry in deer mice.

Influence of hemoglobin-O2 affnity on aerobic capacity in deer mice

High-altitude hypoxia can constrain tissue O2 supply, and several high-altitude populations and species have evolved adaptations to overcome this challenge. Evolved increases in hemoglobin-O2 (Hb-O2) affnity are pervasive across high-altitude taxa, but the influence of such increases on aerobic capacity (maximal O2 consumption) in hypoxia remains contentious. The influence of Hb-O2 affnity on aerobic capacity in hypoxia could vary depending on other traits in the O2 transport pathway, but this possibility is poorly understood. We examined this issue in deer mice ( Peromyscus maniculatus), which is found from sea level to >4300m altitude in the Rocky Mountains. Mice from populations native to high altitude and low altitude were born and raised to adulthood in captivity. Low-altitude mice (n=14) were acclimated to warm (25°C) normoxia, and high-altitude mice (n=14) were acclimated to cold (5°C) hypoxia (~12 kPa O2) for 6 weeks, creating two groups with very different capacities for O2 transport in hypoxia. Aerobic capacity for thermogenesis (VO2max) was then measured in hypoxia after each of three pharmacological treatments to manipulate Hb-O2 affnity: saline (control); efaproxiral, a negative allosteric regulator that decreases Hb-O2 affnity; and sodium cyanate, which covalently modifies Hb and increases Hb-O2 affnity. In control conditions, high-altitude mice had higher VO2max and arterial O2 saturation (SaO2) in hypoxia and higher Hb-O2 affnity (lower P50) than low-altitude mice. Efaproxiral reduced SaO2 and led to similar decreases in VO2max in both populations. Sodium cyanate increased SaO2 in hypoxia in both populations. However, this was only associated with an increase in VO2max in 7 of 14 high-altitude mice and 4 of 14 low-altitude mice. Our results suggest that Hb-O2 affnity may be optimal for aerobic capacity in hypoxia in high-altitude mice, and that reductions in Hb-O2 affnity reduce performance. Supported by NSERC of Canada. This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

Evolved changes in phenotype across skeletal muscles in deer mice native to high altitude.

The cold and hypoxic conditions at high altitude necessitate high metabolic O2 demands to support thermogenesis while hypoxia reduces O2 availability. Skeletal muscles play key roles in thermogenesis, but our appreciation of muscle plasticity and adaptation at high altitude has been hindered by past emphasis on only a small number of muscles. We examined this issue in deer mice (Peromyscus maniculatus). Mice derived from both high-altitude and low-altitude populations were born and raised in captivity and then acclimated as adults to normoxia or hypobaric hypoxia (12 kPa O2 for 6-8 wk). Maximal activities of citrate synthase (CS), cytochrome c oxidase (COX), β-hydroxyacyl-CoA dehydrogenase (HOAD), hexokinase (HK), pyruvate kinase (PK), and lactate dehydrogenase (LDH) were measured in 20 muscles involved in shivering, locomotion, body posture, ventilation, and mastication. Principal components analysis revealed an overall difference in muscle phenotype between populations but no effect of hypoxia acclimation. High-altitude mice had greater activities of mitochondrial enzymes and/or lower activities of PK or LDH across many (but not all) respiratory, limb, core and mastication muscles compared with low-altitude mice. In contrast, chronic hypoxia had very few effects across muscles. Further examination of CS in the gastrocnemius showed that population differences in enzyme activity stemmed from differences in protein abundance and mRNA expression but not from population differences in CS amino acid sequence. Overall, our results suggest that evolved increases in oxidative capacity across many skeletal muscles, at least partially driven by differences in transcriptional regulation, may contribute to high-altitude adaptation in deer mice.NEW & NOTEWORTHY Most previous studies of muscle plasticity and adaptation in high-altitude environments have focused on a very limited number of skeletal muscles. Comparing high-altitude versus low-altitude populations of deer mice, we show that a large number of muscles involved in shivering, locomotion, body posture, ventilation, and mastication exhibit greater mitochondrial enzyme activities in the high-altitude population. Therefore, evolved increases in mitochondrial oxidative capacity across skeletal muscles contribute to high-altitude adaptation.

High-Altitude Hypoxia Exposure Induces Iron Overload and Ferroptosis in Adipose Tissue.

High altitude (HA) has become one of the most challenging environments featuring hypobaric hypoxia, which seriously threatens public health, hence its gradual attraction of public attention over the past decade. The purpose of this study is to investigate the effect of HA hypoxia on iron levels, redox state, inflammation, and ferroptosis in adipose tissue. Here, 40 mice were randomly divided into two groups: the sea-level group and HA hypoxia group (altitude of 5000 m, treatment for 4 weeks). Total iron contents, ferrous iron contents, ROS generation, lipid peroxidation, the oxidative enzyme system, proinflammatory factor secretion, and ferroptosis-related biomarkers were examined, respectively. According to the results, HA exposure increases total iron and ferrous iron levels in both WAT and BAT. Meanwhile, ROS release, MDA, 4-HNE elevation, GSH depletion, as well as the decrease in SOD, CAT, and GSH-Px activities further evidenced a phenotype of redox imbalance in adipose tissue during HA exposure. Additionally, the secretion of inflammatory factors was also significantly enhanced in HA mice. Moreover, the remarkably changed expression of ferroptosis-related markers suggested that HA exposure increased ferroptosis sensitivity in adipose tissue. Overall, this study reveals that HA exposure is capable of inducing adipose tissue redox imbalance, inflammatory response, and ferroptosis, driven in part by changes in iron overload, which is expected to provide novel preventive targets for HA-related illness.

Adaptive increases in respiratory capacity and O2 affinity of subsarcolemmal mitochondria from skeletal muscle of high-altitude deer mice.

Aerobic energy demands have led to the evolution of complex mitochondrial reticula in highly oxidative muscles, but the extent to which metabolic challenges can be met with adaptive changes in physiology of specific mitochondrial fractions remains unresolved. We examined mitochondrial mechanisms supporting adaptive increases in aerobic performance in deer mice (Peromyscus maniculatus) adapted to the hypoxic environment at high altitude. High-altitude and low-altitude mice were born and raised in captivity, and exposed as adults to normoxia or hypobaric hypoxia (12kPa O2 for 6-8weeks). Subsarcolemmal and intermyofibrillar mitochondria were isolated from the gastrocnemius, and a comprehensive substrate titration protocol was used to examine mitochondrial physiology and O2 kinetics by high-resolution respirometry and fluorometry. High-altitude mice had greater yield, respiratory capacity for oxidative phosphorylation, and O2 affinity (lower P50 ) of subsarcolemmal mitochondria compared to low-altitude mice across environments, but there were no species difference in these traits in intermyofibrillar mitochondria. High-altitude mice also had greater capacities of complex II relative to complexes I + II and higher succinate dehydrogenase activities in both mitochondrial fractions. Exposure to chronic hypoxia reduced reactive oxygen species (ROS) emission in high-altitude mice but not in low-altitude mice. Our findings suggest that functional changes in subsarcolemmal mitochondria contribute to improving aerobic performance in hypoxia in high-altitude deer mice. Therefore, physiological variation in specific mitochondrial fractions can help overcome the metabolic challenges of life at high altitude.

Evolved changes in maternal care in high-altitude native deer mice.

At high altitude (HA), unremitting low oxygen and persistent cold push small mammals close to their metabolic ceilings, leaving limited scope for aerobically demanding activities. However, HA breeding seasons are relatively short and endemic rodents compensate with larger litters than low altitude (LA) conspecifics. Rodent mothers are the sole source of heat and nutrition for altricial offspring and lactation is energetically costly. Thus, it is unclear how HA females balance energy allocation during the nursing period. We hypothesized that HA female rodents invest heavily in each litter to ensure postnatal survival. We measured maternal energetic output and behaviour in nursing deer mice (Peromyscus maniculatus) native to LA (400 m a.s.l.) and HA (4350 m a.s.l.) under control (24°C, 760 mmHg) and cold hypoxia conditions, simulating HA (5°C, 430 mmHg). Strikingly, resting metabolic rates of lactating HA and LA females under cold hypoxia were 70-85% of their maximum aerobic capacity. In cold hypoxia, LA mothers increased both nursing time and milk fat content, however their pups were leaner and severely growth restricted at weaning. HA mothers also increased nursing in cold hypoxia but for far less time than LA mothers. Despite receiving less care, HA pups in cold hypoxia only experienced small growth restrictions at weaning and maintained body composition. As adults, HA mice raised in cold hypoxia had increased aerobic capacity compared to controls. These data suggest that HA mothers prioritize their own maintenance costs over investing heavily in their offspring. Pups compensate for this lack of care, likely by reducing their own metabolic costs during development.

Chronic cold exposure induces mitochondrial plasticity in deer mice native to high altitudes.

Small mammals native to high altitude must sustain high rates of thermogenesis to cope with cold. Skeletal muscle is a key site of shivering and non-shivering thermogenesis, but the importance of mitochondrial plasticity in cold hypoxic environments remains unresolved. We examined high-altitude deer mice, which have evolved a high capacity for aerobic thermogenesis, to determine the mechanisms of mitochondrial plasticity during chronic exposure to cold and hypoxia, alone and in combination. Cold exposure in normoxia or hypoxia increased mitochondrial leak respiration and decreased phosphorylation efficiency and OXPHOS coupling efficiency, which may serve to augment non-shivering thermogenesis. Cold also increased muscle oxidative capacity, but reduced the capacity for mitochondrial respiration via complex II relative to complexes I and II combined. High-altitude mice had a more oxidative muscle phenotype than low-altitude mice. Therefore, both plasticity and evolved changes in muscle mitochondria contribute to thermogenesis at high altitude. Small mammals native to high altitude must sustain high rates of thermogenesis to cope with cold and hypoxic environments. Skeletal muscle is a key site of shivering and non-shivering thermogenesis, but the importance of mitochondrial plasticity in small mammals at high altitude remains unresolved. High-altitude deer mice (Peromyscus maniculatus) and low-altitude white-footed mice (P. leucopus) were born and raised in captivity, and chronically exposed as adults to warm (25°C) normoxia, warm hypoxia (12kPa O2 ), cold (5°C) normoxia, or cold hypoxia. We then measured oxidative enzyme activities, oxidative fibre density and capillarity in the gastrocnemius, and used a comprehensive substrate titration protocol to examine the function of muscle mitochondria by high-resolution respirometry. Exposure to cold in both normoxia or hypoxia increased the activities of citrate synthase and cytochrome oxidase. In lowlanders, this was associated with increases in capillary density and the proportional abundance of oxidative muscle fibres, but in highlanders, these traits were unchanged at high levels across environments. Environment had some distinct effects on mitochondrial OXPHOS capacity between species, but the capacity of complex II relative to the combined capacity of complexes I and II was consistently reduced in both cold environments. Both cold environments also increased leak respiration and decreased phosphorylation efficiency and OXPHOS coupling efficiency in both species, which may serve to augment non-shivering thermogenesis. These cold-induced changes in mitochondrial function were overlaid upon the generally more oxidative phenotype of highlanders. Therefore, both plasticity and evolved changes in muscle mitochondria contribute to thermogenesis at high altitudes.

Acclimatization of low altitude-bred deer mice ( Peromyscus maniculatus) to high altitude.

A colony of deer mice subspecies ( Peromyscus maniculatus sonoriensis) native to high altitude (HA) has been maintained at sea level for 18-20 generations and remains genetically unchanged. To determine if these animals retain responsiveness to hypoxia, one group (9-11 wk old) was acclimated to HA (3,800 m) for 8 wk. Age-matched control animals were acclimated to a lower altitude (LA; 252 m). Maximal O2 uptake (V̇o2max) was measured at the respective altitudes. On a separate day, lung volume, diffusing capacity for carbon monoxide (DLCO), and pulmonary blood flow were measured under anesthesia using a rebreathing technique at two inspired O2 tensions. The HA-acclimated deer mice maintained a normal V̇o2max relative to LA baseline. Compared with LA control mice, antemortem lung volume was larger in HA mice in a manner dependent on alveolar O2 tension. Systemic hematocrit, pulmonary blood flow, and standardized DLCO did not differ significantly between groups. HA mice showed a higher postmortem alveolar-capillary hematocrit, larger alveolar ducts, and smaller distal conducting structures. In HA mice, absolute volumes of alveolar type I epithelia and endothelia were higher whereas that of interstitia was lower than in LA mice. These structural changes occurred without a net increase in whole-lung septal tissue-capillary volumes or surface areas. Thus, deer mice bred and raised to adulthood at LA retain phenotypic plasticity and adapt to HA without a decrement in V̇o2max via structural (enlarged airspaces, alveolar septal remodeling) and nonstructural (lung expansion under hypoxia) mechanisms and without an increase in systemic hematocrit or compensatory lung growth. NEW & NOTEWORTHY Deer mice ( Peromyscus maniculatus) are robust and very active mammals that are found across the North American continent. They are also highly adaptable to extreme environments. When introduced to high altitude they retain remarkable adaptive ability to the low-oxygen environment via lung expansion and remodeling of existing lung structure, thereby maintaining normal aerobic capacity without generating more red blood cells or additional lung tissue.

The Mitochondrial Basis for Adaptive Variation in Aerobic Performance in High-Altitude Deer Mice.

Mitochondria play a central role in aerobic performance. Studies aimed at elucidating how evolved variation in mitochondrial physiology contributes to adaptive variation in aerobic performance can therefore provide a unique and powerful lens to understanding the evolution of complex physiological traits. Here, we review our ongoing work on the importance of changes in mitochondrial quantity and quality to adaptive variation in aerobic performance in high-altitude deer mice. Whole-organism aerobic capacity in hypoxia (VO2max) increases in response to hypoxia acclimation in this species, but high-altitude populations have evolved consistently greater VO2max than populations from low altitude. The evolved increase in VO2max in highlanders is associated with an evolved increase in the respiratory capacity of the gastrocnemius muscle. This appears to result from highlanders having more mitochondria in this tissue, attributed to a higher proportional abundance of oxidative fiber-types and a greater mitochondrial volume density within oxidative fibers. The latter is primarily caused by an over-abundance of subsarcolemmal mitochondria in high-altitude mice, which is likely advantageous for mitochondrial O2 supply because more mitochondria are situated adjacent to the cell membrane and close to capillaries. Evolved changes in gastrocnemius phenotype appear to be underpinned by population differences in the expression of genes involved in energy metabolism, muscle development, and vascular development. Hypoxia acclimation has relatively little effect on respiratory capacity of the gastrocnemius, but it increases respiratory capacity of the diaphragm. However, the mechanisms responsible for this increase differ between populations: lowlanders appear to adjust mitochondrial quantity and quality (i.e., increases in citrate synthase [CS] activity, and mitochondrial respiration relative to CS activity) and they exhibit higher rates of mitochondrial release of reactive oxygen species, whereas highlanders only increase mitochondrial quantity in response to hypoxia acclimation. In contrast to the variation in skeletal muscles, the respiratory capacity of cardiac muscle does not appear to be affected by hypoxia acclimation and varies little between populations. Therefore, evolved changes in mitochondrial quantity and quality make important tissue-specific contributions to adaptive variation in aerobic performance in high-altitude deer mice.

Divergent Mitochondrial Antioxidant Activities and Lung Alveolar Architecture in the Lungs of Rats and Mice at High Altitude.

Compared with mice, adult rats living at 3,600 m above sea level (SL—La Paz, Bolivia) have high hematocrit, signs of pulmonary hypertension, and low lung volume with reduced alveolar surface area. This phenotype is associated with chronic mountain sickness in humans living at high altitude (HA). We tested the hypothesis that this phenotype is associated with impaired gas exchange and oxidative stress in the lungs. We used rats and mice (3 months old) living at HA (La Paz) and SL (Quebec City, Canada) to measure arterial oxygen saturation under graded levels of hypoxia (by pulse oximetry), the alveolar surface area in lung slices and the activity of pro- (NADPH and xanthine oxidases—NOX and XO) and anti- (superoxide dismutase, and glutathione peroxidase—SOD and GPx) oxidant enzymes in cytosolic and mitochondrial lung protein extracts. HA rats have a lower arterial oxygen saturation and reduced alveolar surface area compared to HA mice and SL rats. Enzymatic activities (NOX, XO, SOD, and GPx) in the cytosol were similar between HA and SL animals, but SOD and GPx activities in the mitochondria were 2–3 times higher in HA vs. SL rats, and only marginally higher in HA mice vs. SL mice. Furthermore, the maximum activity of cytochrome oxidase-c (COX) measured in mitochondrial lung extracts was also 2 times higher in HA rats compared with SL rats, while there was only a small increase in HA mice vs. SL mice. Interestingly, compared with SL controls, alterations in lung morphology are not observed for young rats at HA (15 days after birth), and enzymatic activities are only slightly altered. These results suggest that rats living at HA have a gradual reduction of their alveolar surface area beyond the postnatal period. We can speculate that the elevation of SOD, GPx, and COX activities in the lung mitochondria are not sufficient to compensate for oxidative stress, leading to damage of the lung tissue in rats.

Evolved changes in the intracellular distribution and physiology of muscle mitochondria in high‐altitude native deer mice

Mitochondrial function changes over time at high altitudes, but the potential benefits of these changes for hypoxia resistance remains unclear. We used high-altitude-adapted populations of deer mice, which exhibit enhanced aerobic performance in hypoxia, to examine whether changes in mitochondrial physiology or intracellular distribution in the muscle contribute to hypoxia resistance. Permeabilized muscle fibres from the gastrocnemius muscle had higher respiratory capacities in high-altitude mice than in low-altitude mice. Highlanders also had higher mitochondrial volume densities, due entirely to an enriched abundance of subsarcolemmal mitochondria, such that more mitochondria were situated near the cell membrane and adjacent to capillaries. There were several effects of hypoxia acclimation on mitochondrial function, some of which were population specific, but they differed from the evolved changes in high-altitude natives, which probably provide a better indication of adaptive traits that improve performance and hypoxia resistance at high altitudes. High-altitude natives that have evolved to live in hypoxic environments provide a compelling system to understand how animals can overcome impairments in oxygen availability. We examined whether these include changes in mitochondrial physiology or intracellular distribution that contribute to hypoxia resistance in high-altitude deer mice (Peromyscus maniculatus). Mice from populations native to high and low altitudes were born and raised in captivity, and as adults were acclimated to normoxia or hypobaric hypoxia (equivalent to 4300m elevation). We found that highlanders had higher respiratory capacities in the gastrocnemius (but not soleus) muscle than lowlanders (assessed using permeabilized fibres with single or multiple inputs to the electron transport system), due in large part to higher mitochondrial volume densities in the gastrocnemius. The latter was attributed to an increased abundance of subsarcolemmal (but not intermyofibrillar) mitochondria, such that more mitochondria were situated near the cell membrane and adjacent to capillaries. Hypoxia acclimation had no significant effect on these population differences, but it did increase mitochondrial cristae surface densities of mitochondria in both populations. Hypoxia acclimation also altered the physiology of isolated mitochondria by affecting respiratory capacities and cytochrome c oxidase activities in population-specific manners. Chronic hypoxia decreased the release of reactive oxygen species by isolated mitochondria in both populations. There were subtle differences in O2 kinetics between populations, with highlanders exhibiting increased mitochondrial O2 affinity or catalytic efficiency in some conditions. Our results suggest that evolved changes in mitochondrial physiology in high-altitude natives are distinct from the effects of hypoxia acclimation, and probably provide a better indication of adaptive traits that improve performance and hypoxia resistance at high altitudes.

BIRTH ALTITUDE AFFECTS AEROBIC PERFORMANCE

High altitude rarefied atmospheres can come as a shock if you're used to life at sea-level. No matter how hard you breathe, initially you're left gasping. However, given time, most species adjust to the new conditions. Gregory Russell from the University of California at Riverside explains that animals living at altitude tend to have larger hearts and lungs than their sea-level relatives. Yet despite this advantage, they don't always perform better than mice living at lower altitudes, possibly due to low oxygen levels limiting the animal's aerobic performance. Which made Russell, Enrico Rezende and Kimberly Hammond wonder how much of an effect developmental history has on an animal's ability to cope with different oxygen levels. The team decided to see how the aerobic performance of adult deer mice was affected by the oxygen level and altitude of their birth(p. 35).With access to a lab-based colony of deer mice, Russell transported twenty mouse pairs to 3800 m altitude at the White Mountain Research Station ready to see how their high-born young faired relative to low-born mice. At 5 weeks of age, Russell put each of the high- and low-altitude-born mice in a treadmill and measured their aerobic performances. Then he artificially reduced the oxygen levels in the treadmill for the low-altitude mice (to simulate the effects of hypoxia at high altitude) and increased the oxygen for the high-altitude mice (simulating the effects of normoxia at low altitude) to see how they faired. The animals born in low oxygen (hypoxia) at high altitude out-performed the mice born in high oxygen (normoxia) at low altitude. At 5 weeks of age it seemed as if being born at high altitude had given the mice an advantage over the low-altitude pups.So how would the pups born at high and low altitude adjust when transported to the opposite situation? Russell carefully transported half of the 5-week-old high-altitude pups down to the Riverside labs, and half of the Riverside pups up to the mountains, leaving the other two groups at their birth places. Giving the relocated animals 8 weeks to acclimate to their new environment, Russell again tested all of the animals' aerobic performances in response to hypoxia and normoxia.Amazingly, pups born at low altitude (normoxia) outperformed all of the other mice after a period of acclimation to high altitude, while the performance of pups born at high altitude (hypoxia) changed little or declined after remaining at altitude or acclimating at low altitude. The team realised that when they had tested the 5-week-old animals, the high-altitude pups must have already acclimated to the hypoxic conditions. And when the team analysed the animals' aerobic performances, it became clear to them that there must be a physiological difference between pups gestated and born at high altitude,and pups born at low altitude. `How the mice accommodate the challenges imposed by high altitude depends on where they are born' says Russell. `We don't know how they do it', he adds `but suspect that high-altitude mice acclimate in a different way'.

Altitudinal Effects on Water Fluxes of Deer Mice: A Physiological Application of Structural Equation Modeling with Latent Variables

An animal's physiological capacities, the environment it inhabits, and their interaction determine the animal's water flux (turnover rate). The proximate physiological determinants of water flux (e.g., urinary concentrating ability, evaporative water loss, dietary water content, and oxidative water yield) are well understood. Excluding effects of diet type (e.g, herbivory vs. granivory) and free water availability (e.g., xeric vs. mesic habitats), ecological and environmental effects on water fluxes are less well known. We used labeled water to study the effects of large altitudinal differences (~ 3,000 m) on water fluxes of free-living deer mice. In absolute terms, high-altitude mice had higher water fluxes and field metabolic rates, but were smaller, than low-altitude mice. A multiple regression with altitude coded as a dummy variable indicated that after adjusting for mass and field metabolic rate, altitude had no effect on water flux. However, using mass and field metabolic rate as independent variabl...

Muscle capillary tortuosity in high altitude mice depends on sarcomere length

At given capillary-to-fiber ratio and sarcomere length, capillary tortuosity determines capillary length and the geometry of blood-tissue exchange in muscles. It is therefore an important determinant of the potential efficiency of O 2 supply to the muscle fibers. Reports have indicated that capillary tortuosity increases with adaptation to hypoxia. Unfortunately, sarcomere length was not taken into account. Because capillary tortuosity increases substantially as the muscle shortens, it is important to measure sarcomere length when comparing capillary configuration among muscles. We addressed the question of whether or not capillary tortuosity is greater in skeletal muscles of high altitude mice compared to sea-level, when account is taken of sarcomere length. Calf and thigh muscles of high altitude (3800 m; inspired P O2, 91 mm Hg) and sea-level deer mice, Peromyscus maniculatus, were perfusion-fixed in situ at sarcomere lengths ranging from 1.9 to 2.4 μm. Capillary-to-fiber ratio, capillary length per volume of fiber, and the degree of orientation (anisotropy) of capillaries were estimated by morphometry. In both sea-level and high altitude mice, capillaries were relatively straight in extended muscles, and substantially more tortuous in muscles fixed at shorter sarcomere length. There was no systematic difference in the degree of tortuosity of capillaries in the high altitude compared to sea-level mice, when account was taken of sarcomere length. Capillary length per volume of muscle fiber and capillary-to-fiber ratio were not significantly different between the two groups.

Altitudinal and seasonal effects on aerobic metabolism of deer mice.

I compared the maximal aerobic metabolic rates (VO2max), field metabolic rates (FMR), aerobic reserves (VO2max-FMR), and basal metabolic rates (BMR) of wild and recently captured deer mice from low (440 m) and high (3800 m) altitudes. To separate the effects of the thermal environment from other altitudinal effects, I examined mice from different altitudes, but similar thermal environments (i.e., summer mice from high altitude and winter mice from low altitude). When the thermal environment was similar, VO2max, FMR, and aerobic reserve of low and high altitude mice did not differ, but BMR was significantly higher at high altitude. Thus, in the absence of thermal differences, altitude had only minor effects on the aerobic metabolism of wild or recently captured deer mice. At low altitude, there was significant seasonal variation in VO2max, FMR, and aerobic reserve, but not BMR. BMR was correlated with VO2max, but not with FMR. The significant positive correlation of BMR with VO2max indicates a cost of high VO2max, because higher BMR increases food requirements and energy use during periods of thermoneutral conditions.

A stereological analysis of the ultrastructure of the lungs of wild mice living at low and high altitude.

Stereological analysis of the ultrastructural composition of the pulmonary alveolo-capillary region of mice living at sea level compared with that of the same species (Phyllotis darwini) genetically adapted to life at 4,660 m reveals a trend at high altitude towards a greater volume percentage of tissue components. On a weight-specific basis, non-circulating tissue occupies a significantly greater volume in high-altitude mice, but air space and capillary contents are not correspondingly greater. Since the arithmetic mean thickness of the tissue layers and of the air-blood barrier are the same in the two altitudinal groups, the average alveolus must have a smaller volume in the high-altitude mice. Epithelial, endothelial, and erythrocyte surface areas per gram body weight are significantly greater in the high-altitude mice. Nuclear counts indicate that the larger lungs of mice adapted to high altitude are due to larger Pneumocyte I and II and endothelial cells rather than to an increase in the number of these cells. Hematocrits measured within the pulmonary capillaries in the two altitudinal groups were equal. An heretofore unrecognized feature of possible adaptive value is the surface/volume ratio of erythrocytes, which is similar for erythrocytes in alveolar space of mice at low and high altitudes but within lung capillaries is 14.7% greater at high altitude.

Influence of altitude on heart and breathing rates in some Peruvian rodents.

Heart and breathing rates were measured at 0.1 and 4.5 km before, during, and after Nembutal anesthesia in Phyllotis darwini chilensis, P. d. rupestris, P. osilae, P. pictus, Akodon amoenus, A. berlepschii, A. jelskii, and A. boliviensis; in Mus musculus rates were measured at 3.9-4.5 km, and in P. d. limatus, M. musculus, and the white mouse at 0.1 km. None of the high-altitude mice and only the native Phyllotis (+25%) of the low-altitude mice showed a circulatory response to altitude. All the low-altitude mice but only Akodon (+22%) of the high-altitude mice showed a respiratory response to altitude. The native mountain species had higher resting heart and breathing rates than did the lowland forms. Submitted on October 25, 1961