[Phylogeny of gas exchange systems]. / Phylogenese der Gasaustauschsysteme.

Several systems of gas transport have developed during evolution, all of which are able to sufficiently supply oxygen to the tissues and eliminate the CO2 produced by the metabolism, in spite of great distances between the environment and the individual cells of the tissues. Almost all these systems utilize a combination of convection and diffusion steps. Convection achieves an efficient transport of gas over large distances, but requires energy and cannot occur across tissue barriers. Diffusion, on the other hand, achieves gas transport across barriers, but requires optimization of diffusion paths and diffusion areas. When two convectional gas flows are linked via a diffusional barrier (gas/fluid in the case of the avian lung, fluid/fluid in the case of gills), the directions in which the respective convectional movements pass each other are important determinants of gas exchange efficiency (concurrent, countercurrent and cross-current systems). The tracheal respiration found in insects has the advantage of circumventing the convective gas transport step in the blood, thereby avoiding the high energy expenditure of circulatory systems. This is made possible by a system of tracheae, ending in tracheoles, that reaches from the body surface to every cell within the body. The last step of gas transfer in these animals occurs by diffusion from the tracheoles ("air capillaries") to the mitochondria of cells. The disadvantage is that the tracheal system occupies a substantial fraction of body volume and that, due to limited mechanical stability of tracheal walls, this system would not be able to operate under conditions of high hydrostatic pressures, i. e. in large animals. Respiration in an "open" system, i. e. direct exposure of the diffusional barrier to the environmental air, eliminates the problem of bringing the oxygen to the barrier by convection, as is necessary in the avian and mammalian lung, in the insects' tracheal system and in the gills. An open system is found in the respiration via the skin, which is of significance in some amphibians, but is limited by the thickness of the skin that constitutes a substantial diffusion path for O2 and CO2. The thick skin, on the other hand, provides mechanical protection as well as flexibility for the animals' body and helps avoid massive water loss via the body surface. The gills of fishes, in contrast, exhibit rather short diffusion distances, are located in a mechanically protected space, and the problem of water loss does not exist. The flows of blood and water occur in opposite direction (countercurrent flow) and this situation makes an arterial PO2 approaching the environmental PO2 possible. A major disadvantage is constituted by the environmental medium since water contains little O2 compared to air and, to compensate this, much energy is expended to maintain a high flow rate of water through the gills. In the mammalian lung ("pool system"), the presence of a dead space and the rhythmic ventilation that replaces only a small fraction of the gas volume of the lung per breath, are responsible for an arterial PO2 (2/3 of the atmospheric PO2) that cannot reach the expiratory PO2. However, an advantage of this feature is the constantly high alveolar and arterial PCO2, which provides a highly effective H(+) buffer system in the entire body. The apparent disadvantage of the mammalian lung is avoided by the avian lung, which uses an extended system of airways to establish continuous equilibration of a part of the capillary blood with fresh air (cross current system), during inspiration as well as during expiration. In this system, arterial PO2 can significantly exceed expiratory PO2. A disadvantage here is the enormous amount of space taken up by the avian lung, in animals of 1 kg body weight three times as much as taken up by the mammalian lung. All respiratory exchange systems considered here exhibit high degrees of optimization - yet follow highly diverse construction principles. There is no such thing as an ideal gas exchange system. The system that has evolved in each species depends to an impressive extent on environmental conditions, on body build and size, on the animal's patterns of movement and on its energy consumption.

Radial and longitudinal diffusion of myoglobin in single living heart and skeletal muscle cells.

We have used a fluorescence recovery after photobleaching (FRAP) technique to measure radial diffusion of myoglobin and other proteins in single skeletal and cardiac muscle cells. We compare the radial diffusivities, D(r) (i.e., diffusion perpendicular to the long fiber axis), with longitudinal ones, D(l) (i.e., parallel to the long fiber axis), both measured by the same technique, for myoglobin (17 kDa), lactalbumin (14 kDa), and ovalbumin (45 kDa). At 22 degrees C, D(l) for myoglobin is 1.2 x 10(-7) cm(2)/s in soleus fibers and 1.1 x 10(-7) cm(2)/s in cardiomyocytes. D(l) for lactalbumin is similar in both cell types. D(r) for myoglobin is 1.2 x 10(-7) cm(2)/s in soleus fibers and 1.1 x 10(-7) cm(2)/s in cardiomyocytes and, again, similar for lactalbumin. D(l) and D(r) for ovalbumin are 0.5 x 10(-7) cm(2)/s. In the case of myoglobin, both D(l) and D(r) at 37 degrees C are about 80% higher than at 22 degrees C. We conclude that intracellular diffusivity of myoglobin and other proteins (i) is very low in striated muscle cells, approximately 1/10 of the value in dilute protein solution, (ii) is not markedly different in longitudinal and radial direction, and (iii) is identical in heart and skeletal muscle. A Krogh cylinder model calculation holding for steady-state tissue oxygenation predicts that, based on these myoglobin diffusivities, myoglobin-facilitated oxygen diffusion contributes 4% to the overall intracellular oxygen transport of maximally exercising skeletal muscle and less than 2% to that of heart under conditions of high work load.

Protein diffusion in living skeletal muscle fibers: dependence on protein size, fiber type, and contraction.

Sarcoplasmic protein diffusion was studied under different conditions, using microinjection in combination with microspectrophotometry. Six globular proteins with molecular masses between 12 and 3700 kDa, with diameters from 3 to 30 nm, were used for the experiments. Proteins were injected into single, intact skeletal muscle fibers taken from either soleus or extensor digitorum longus (edl) muscle of adult rats. No correlation was found between sarcomere spacing and the sarcoplasmic diffusion coefficient (D) for all proteins studied. D of the smaller proteins cytochrome c (diameter 3.1 nm), myoglobin (diameter 3.5 nm), and hemoglobin (diameter 5.5 nm) amounted to only approximately 1/10 of their value in water and was not increased by auxotonic fiber contractions. D for cytochrome c and myoglobin was significantly higher in fibers from edl (mainly type II fibers) compared to fibers from soleus (mainly type I fibers). Measurements of D for myoglobin at 37 degrees C in addition to 22 degrees C led to a Q(10) of 1.46 for this temperature range. For the larger proteins catalase (diameter 10.5 nm) and ferritin (diameter 12.2 nm), a decrease in D to approximately 1/20 and approximately 1/50 of that in water was observed, whereas no diffusive flux at all of earthworm hemoglobin (diameter 30 nm) along the fiber axis could be detected. We conclude that 1) sarcoplasmic protein diffusion is strongly impaired by the presence of the myofilamental lattice, which also gives rise to differences in diffusivity between different fiber types; 2) contractions do not cause significant convection in sarcoplasm and do not lead to increased diffusional transport; and 3) in addition to the steric hindrance that slows down the diffusion of smaller proteins, diffusion of large proteins is further hindered when their dimensions approach the interfilament distances. This molecular sieve property progressively reduces intracellular diffusion of proteins when the molecular diameter increases to more than approximately 10 nm.

Contraction parameters, myosin composition and metabolic enzymes of the skeletal muscles of the etruscan shrew Suncus etruscus and of the common European white-toothed shrew Crocidura russula (Insectivora: soricidae).

In the Etruscan shrew, the isometric twitch contraction times of extensor digitorum longus (EDL) and soleus muscles are shorter than in any other mammal, allowing these muscles to contract at outstandingly high contraction frequencies. This species has the highest mass-specific metabolic rate of all mammals and requires fast skeletal muscles not only for locomotion but also for effective heat production and for an extremely high ventilation rate. No differences could be detected in the fibre type pattern, the myosin heavy and light chain composition, or in the activity of the metabolic enzymes lactate dehydrogenase and citrate synthase of the two limb muscles, the EDL and the soleus, which in larger mammalian species exhibit distinct differences in contractile proteins and metabolic enzymes. All properties determined in EDL and soleus muscles of Suncus etruscus, as well as in the larger Crocidura russula, are typical for fast-oxidative fibres, and the same holds for several other skeletal muscles including the diaphragm muscle of S. etruscus. Nevertheless, the EDL and soleus muscles showed different mechanical properties in the two shrew species. Relaxation times and, in C. russula, time to peak force are shorter in the EDL than in the soleus muscle. This is in accordance with the time course of the Ca(2+) transients in these muscles. Such a result could be due to different parvalbumin concentrations, to a different volume fraction of the sarcoplasmic reticulum in the two muscles or to different Ca(2+)-ATPase activities. Alternatively, the lower content of cytosolic creatine kinase (CK) in the soleus compared with the EDL muscle could indicate that the observed difference in contraction times between these shrew muscles is due to the CK-controlled activity of their sarcoplasmic reticulum Ca(2+)-ATPase.

Rates of rewarming, heart and respiratory rates and their significance for oxygen transport during arousal from torpor in the smallest mammal, the Etruscan shrew Suncus etruscus.

We investigated the process of rewarming from torpor with respect to respiratory and circulatory oxygen transport properties in the smallest mammal, the Etruscan shrew Suncus etruscus. In seven adult Etruscan shrews with a mean body mass of 2.4g, torpor was induced by deprivation of food and a cold environment. During arousal from torpor at an ambient temperature of 22 degrees C, the shrews actively rewarmed from the lowest mean (+/- S.D.) body temperature (Tb) of 12.1 +/- 1.2 degrees C to 20 degrees C at a rate of 0.43 +/- 0.14 degree C min-1, from 20 to 24 degrees C at a rate of 0.8 degree C min-1, and from 24 to 36 degrees C at a rate of 1.1 +/- 0.1 degrees C min-1. The mean rate from 12 degrees C to normothermia amounted to 0.83 degree C min-1, which is among the highest values recorded in mammals. During rewarming, the heart rate increased exponentially (Q10 = 2.2) from 100 to 800-1200 min-1, whereas the respiratory rate increased linearly from 50 to 600-800 min-1. These rates are higher than the heart and respiratory rates reported for other small mammals at the same Tb. The fraction of brown adipose tissue (BAT) was 9.2 +/- 1.6% of body mass, which is higher than in any other mammal. Up to a body temperature of approximately 17 degrees C, the heat for rewarming was mainly produced in the BAT; above this value, considerable activity of the skeletal muscles enhanced thermogenesis. Estimation of the mixed venous oxygen partial pressure showed that, at the tissue level, the rewarming process corresponds to heavy work conditions. The ventilatory system is adapted such that during rewarming, in addition to the appropriate oxygen transport capacity, there is also a capacity for hyperventilation.

Heart and respiratory rates and their significance for convective oxygen transport rates in the smallest mammal, the Etruscan shrew Suncus etruscus.

Heart and respiratory rates of the smallest mammal (mean adult body mass 2g), the Etruscan shrew Suncus etruscus, were determined at rest and under stress conditions. Heart rate was obtained from electrocardiograms (ECGs), recorded via foot electrodes. The mean +/- S.D. heart rate of resting animals (ambient temperature 22 degrees C) was 835 +/- 107 min-1, the mean maximal rate amounted to 1093 +/- 235 min-1. The highest single value recorded was 1511 min-1, which is the highest heart rate reported so far for an endotherm. The respiratory rate was also obtained from ECG recordings, which showed the electrical activity of the breathing muscles during inhalation, and additionally by recording the movements of the thoracic wall with a laser autofocus system. The mean resting respiratory rate was 661 +/- 93 min-1, the mean maximal rate was 758 +/- 109 min-1 and the highest single value recorded was 894 min-1. At 22 degrees C, the specific oxygen consumption rate is 67 times higher in resting S. etruscus than in resting humans. Under these conditions, the respiratory rate of the shrew is 47 times higher but the heart rate only 12 times higher than in man. Therefore, to achieve an adequate circulatory oxygen transport rate, the product of relative stroke volume and arterio-venous O2 difference has to be 5.6 times higher in the shrew than in man, whereas for an appropriate ventilatory oxygen transport rate the product of relative tidal volume and oxygen extraction has to be only 1.4 times higher in this small insectivore than in man. The maximal possible oxygen transport rates of the ventilatory and the circulatory system have been estimated and compared with the diffusional transport capacity of the lung. These rates amount to approximately 1000 ml O2 kg-1 min-1. According to our results and data in the literature, an aerobic scope of 7-10 seems to be realistic for the Etruscan shrew.

Diffusion of myoglobin in skeletal muscle cells--dependence on fibre type, contraction and temperature.

We measured the diffusion coefficient of myoglobin (DMb) inside mammalian skeletal muscle cells with a microinjection technique. A small bolus of horse Mb was injected into a single muscle fibre and the subsequent time-dependent changes of the Mb profiles along the fibre axis were measured with a microscope-photometer. For fibres of the rat soleus muscle at 22 degrees C, a DMb of 1.3.10(-7) cm2/s was found, confirming a result obtained previously by us for rat diaphragm muscle with a photo-oxidation technique. In the extensor digitorum longus muscle of the rat, a higher value of 1.9.10(-7) cm2/s was measured. Auxotonic muscle contractions did not change the apparent DMb. For the temperature range between 22 degrees C and 37 degrees C, a temperature coefficient. Q10, of 1.5 was calculated. The implication of this result for the role of Mb in the facilitation of oxygen transport was examined. Model calculations show that with this relatively low DMb value, the intracellular oxygen supply can be improved only slightly.

[The value of myocardia protection in chronic hypoxic immature rat hearts]. / Die Wertigkeit myokardprotektiver Methoden bei chronisch hypoxischen unreifen Rattenherzen.

This study examines the efficacy of three methods for myocardial protection during 8 hours of global ischema at 10 degrees C in immature (28 days) rat hearts subjected to lifelong hypoxia afforded by exposure to simulated high altitude. Hearts in group 1 were protected by rapid topical cooling alone, in group 2 by slow pre-arrest cooling with Krebs-Henseleit solution plus topical cooling and in group 3 by coronary perfusion with St. Thomas' Hospital cardioplegia No 2 (STS 2) plus topical cooling. Hearts in groups 4-6 served as controls without hypoxia and were protected accordingly. Parameters of myocardial function (left ventricular pressure, LVP), the metabolic status (myocardial concentration of ATP and creatine phosphate) and endothelial function (response to the vasodilator acetylcholine) were measured. Myocardial protection by rapid topical cooling alone resulted in equal--or significantly improved--postischemic recovery of LVP and endothelial function compared with slow pre-arrest cooling or additional protection with STS 2. The data advocate topical cooling for myocardial protection during surgical correction of cyanotic congenital cardiac disease in early infancy. In this age group, coronary perfusion with cold crystalloid solutions appears to aggravate ischemic endothelial injury.

Diffusivity of myoglobin in intact skeletal muscle cells.

We report a method that allows us to determine the diffusion coefficient of native myoglobin in intact and mechanically unaffected red muscle fibers. The method is based on an optical recording of intracellular diffusion of metmyoglobin, which is produced inside the cells by photooxidation of oxymyoglobin with a UV light pulse. We find a myoglobin diffusivity of 1.2 x 10(-7) cm2/s (22 degrees C), which is only 1/10th of the value measured in very dilute myoglobin solutions and 1/5th of the value obtained from measurements in solutions of myoglobin at 18 g/dl. The latter value often has been used in model calculations of oxygen transport to tissue incorporating myoglobin-facilitated oxygen diffusion. Recalculating facilitated diffusion with the value obtained by us implies that its contribution to total intracellular oxygen transport is of minor importance. Furthermore, it shows that sterical hindrance to myoglobin diffusion is dominated by the muscle-cell architecture rather than by the overall protein concentration of the muscle fiber.

Lifetime energy budgets in mammals and birds.

1. Two data sets for standard energy metabolism (351 and 320 species, respectively) and one for maximal lifespan (494 species) in mammals have been assembled from the literature. 2. In addition smaller data sets of active (field) energy metabolism in mammals (36 species) and in birds (25 species) have been drawn on. 3. The products of the respective regression parameters as well as the products of energy metabolism and maximal lifespan in individual species have been computed in order to estimate lifetime energy metabolism in mammals generally and in various mammalian orders. 4. It is found that lifetime energy budgets in mammals generally, whether standard or active, very systematically with body mass with slopes between 0.87 and 0.93, significantly different from unity (P less than 0.001 or P less than 0.01). 5. In birds, lifetime energy budgets, whether standard or active, vary with slopes of 0.94 +/- 0.05 and 0.88 +/- 0.09, which are not significantly different from unity (P greater than 0.1). 6. In carnivores, artiodactyls, primates and bats the slopes for lifetime standard as well as lifetime active energy budgets are not significantly different from one in any of the investigated data sets. 7. In rodents the lifetime standard energy budgets have slope significantly different from one; in marsupials one data set for lifetime standard and the one for lifetime active energy budget lead to slopes significantly different from one. 8. It is concluded from this analysis that current data do not support the hypothesis that lifetime energy budgets, whether standard or active, vary as the first power of body mass in mammals generally.(ABSTRACT TRUNCATED AT 250 WORDS)

Primary structure and oxygen-binding properties of the hemoglobin from guanaco (Lama guanacoë, Tylopoda).

The primary structure of the hemoglobin from guanaco (Lama guanacoë, Tylopoda) is presented. It could be separated into the chains by CM-cellulose chromatography. The sequences have been determined by automatic Edman degradation with the film technique or gas phase method, using the native chains and the tryptic peptides of the oxidized chains as well as a fragment obtained by acid hydrolysis. Guanaco hemoglobin has identical alpha-chains with alpaca and identical beta-chains with all Lama species with the exception that one guanaco in this study had alanine and serine in the ratio 1:1 in position beta 135 whereas a second individual had alanine only. Since the data suggest that the domesticated species llama and alpaca originate from the guanaco, it seems likely that beta 135Ala is the common form. Guanaco, llama, and alpaca show a comparable high blood oxygen affinity, caused by the substitution beta 2(NA2)His----Asn, as an adaptation to life at high altitude.

[Strategies of adaptation of oxygen transport systems in mammals to life at high altitude]. / Strategien der Anpassung des Sauerstofftransportsystems von Säugetieren an das Leben in grossen Höhen.

Like many mammalian species, man responds to exposure to high altitude with an increasing hemoglobin concentration and a decreasing blood O2-affinity. The Lama vicugna of the Andes does not show this kind of adaptation to altitude. Even at high altitude its hematocrit is unusually low and its O2-affinity is the highest known among mammals. Additionally, it is characterized by a high capillary density of the muscles and a large relative heart weight. The consequences of these different strategies of altitude adaptation of the oxygen supply system and the capability for physical performance are discussed.

Oxygen binding properties, capillary densities and heart weights in high altitude camelids.

The oxygen binding properties of the blood of the camelid species vicuna, llama, alpaca and dromedary camel were measured and evaluated with respect to interspecific differences. The highest blood oxygen affinity, not only among camelids but of all mammals investigated so far, was found in the vicuna (P50 = 17.6 Torr compared to 20.3-21.6 Torr in the other species). Low hematocrits (23-34%) and small red blood cells (21-30 microns 3) are common features of all camelids, but the lowest values are found in the Lama species. Capillary densities were determined in heart and soleus muscle of vicuna and llama. Again, the vicuna shows exceptional values (3720 cap/mm2 on average in the heart) for a mammal of this body size. Finally, heart weight as percent of body weight is higher in the vicuna (0.7-0.9%) than in the other camelids studied (0.5-0.7%). The possibility that these parameters, measured in New World tylopodes at sea level, are not likely to change considerably with transfer to high altitude, is discussed. In the vicuna, a unique combination of the following features seems to be responsible for an outstanding physical capability at high altitude: saturation of blood with oxygen in the lung is favored by a high blood oxygen affinity, oxygen supply being facilitated by low diffusion distances in the muscle tissue. Loading, as well as unloading, of oxygen is improved by a relatively high oxygen transfer conductance of the red blood cells, which is due to their small size and which compensates the negative effect of a low hematocrit on the oxygen conductance of blood.(ABSTRACT TRUNCATED AT 250 WORDS)

Dependence of O2 transfer conductance of red blood cells on cellular dimensions.

To estimate the significance of the dimensions of RBC on O2 transfer, the kinetics of O2 release from RBC into medium containing dithionite (40 mmol/l) was measured, by a stopped-flow technique, for nine different species with varying RBC size (man, llama, vicuna, alpaca, dromedary camel, pygmy goat, domestic hen, muscovy duck and turtle). The observed O2 transfer kinetics were found to be size-dependent, i.e. the O2 transfer conductance of the single RBC, gst, was lower, whereas the specific O2 transfer conductance of packed RBC, Gst, or of whole blood, theta st, was higher for smaller RBC. The ratio of surface area to effective diffusion path length which was found to be about one fourth of the mean cell thickness irrespective of cell size and cell shape, may be considered as the essential morphological factor determining O2 transfer efficiency of the single RBC.

Oxygen transfer properties and dimensions of red blood cells in high-altitude camelids, dromedary camel and goat.

To estimate the advantage of the small red blood cells (RBC) of high-altitude camelids for O2 transfer, the kinetics of O2 uptake into and release from the RBC obtained from llama, vicuña and alpaca were investigated at 37 degrees C with a stopped-flow technique. O2 transfer conductance of RBC (G) was estimated from the rate of O2 saturation change and the corresponding O2 pressure difference between medium and hemoglobin. For comparison, O2 kinetics for the RBC of a low-altitude camelid (dromedary camel) and the pygmy goat were determined and previously measured values for human RBC were used. O2 transfer of RBC was found to be strongly influenced by extracellular diffusion, except with O2 release into dithionite solutions of sufficiently high concentration (greater than 30 mM). The G values measured in these 'standard' conditions, Gst (in mmol X min-1 X Torr-1 X (ml RBC)-1) were: high-altitude camelids, 0.58 (averaged for llama, alpaca and vicuña since there were no significant interspecific differences); camel 0.42; goat, 0.42; man, 0.39. The differences can in part be attributed to expected effects of the size and shape of the RBC (volume, surface area, mean thickness), as well as to the intracellular O2 diffusivity which depends on the concentration of cellular hemoglobin. The high Gst of RBC of high-altitude camelids may be considered to enhance O2 transfer in lungs and tissues. But the O2 transfer conductance of blood, theta, equal to Gst multiplied by hematocrit (in mmol X min-1 X Torr-1 X (ml blood)-1), was only slightly higher as compared to other species: 0.20 (llama, alpaca, vicuña), 0.14 (camel), 0.18 (goat), 0.17 (man).

Scaling of maximal lifespan in bats.

1. Values for maximal lifespan in heterothermic and homeothermic bats as a function of body weight, brain weight and lifetime basal energy consumption were submitted to linear (log-log) and multiple regression analysis. 2. The results of the regression analyses of maximal lifespan in bats were compared with those reported for non-flying mammals based on both narrow and wide weight ranges. 3. It was found that the regression lines (linear or multiple) for maximal lifespan in bats (heterothermic or homeothermic) lie well above the regression lines for non-flying mammals. 4. Predictions of maximal lifespan in heterothermic bats based on estimated lifetime basal energy consumption and body weight are in reasonable agreement with observed values when torpor and hibernation behaviour are taken into account. 5. But observed values of maximal lifespan in homeothermic bats were found to lie substantially above the regression lines derived for non-flying mammals. 6. It was concluded that existing hypotheses do not account for the long lifespan observed in bats generally.