Hemorheology and oxygen transport in vertebrates. A role in thermoregulation?

We studied the effect of temperature on blood rheology in three vertebrate species with different thermoregulation and erythrocyte characteristics. Higher fibrinogen proportion to total plasma protein was found in turtles (20%) than in pigeons (5.6%) and rats (4.2%). Higher plasma viscosity at room temperature than at homeotherm body temperature was observed in rats (1.69 mPa x s at 20 degrees C vs. 1.33 mPa x s at 37 degrees C), pigeons (3.40 mPa x s at 20 degrees C vs. 1.75 mPa x s at 40 degrees C), and turtles (1.74 mPa x s at 20 degrees C vs. 1.32 mPa x s at 37 degrees C). This fact allow us to hypothesize that thermal changes in protein structure may account for an adjustment of the plasma viscosity. Blood viscosity was dependent on shear rate, temperature and hematocrit in the three species. A different behaviour in apparent and relative viscosities between rat and pigeon at environmental temperature was found. Moreover, the blood oxygen transport capacity seems more affected by a reduction of temperature in rats than in pigeons. Both findings indicate a greater influence of temperature on mammalian erythrocyte than on nucleated red cells, possibly as a consequence of differences in thermal sensitivity and mechanical stability between them. A comparison between the three species revealed that apparent blood viscosity measured at homeotherm physiological temperature was linearly related to the hematocrit level of each species. However, when measured at environmental temperature, rat blood showed a higher apparent viscosity than those found in species with non-nucleated red cells, thus indicating a higher impact of temperature decrease on blood viscosity in mammals. This suggest that regional hypothermia caused by cold exposure may affect mammalian blood rheological behaviour in a higher extent than in other vertebrate species having nucleated red cells and, consequently, influencing circulatory function and oxygen transport.

Capillarity, fibre types and fibre morphometry in different sampling sites across and along the tibialis anterior muscle of the rat.

Capillarity, fibre types, fibre cross-sectional areas and perimeters were studied along and across the rat tibialis anterior muscle. The muscle was sectioned at three different levels (proximal, equatorial and distal) choosing five sampling fields for measurements at each level (from anterior to posterior and lateral to medial zones). Significant differences were found in the percentage of fibre types and capillarity between different fields of the same muscle section. Slow oxidative fibres were confined to the posterior muscle zone with a maximum of 3.7%. The posterior fields also had a greater percentage of fast oxidative glycolytic fibres at proximal (72.3%) and equatorial (61.3%) levels, but a lower value at the distal level (44.8%) and lower capillary density counts in total cross-section means (758 vs. 1,069 capillaries/mm(2) in equatorial and 1,035 capillaries/mm(2) at proximal levels). The uneven distribution of both fast fibre types and the different degrees of capillarisation along the muscle are statistically significant and may be due to different biomechanical performances along the rat tibialis anterior. Fibre size was significantly larger at the distal level, but no morphometric differences were found across the section of the same level. At the distal level, the mean total fibre area of fast glycolytic fibres (5,130 microm(2)) and fast oxidative glycolytic fibres (2,493 microm(2)) contrasted with values at the proximal (fast glycolytic: 4,070 microm(2), fast oxidative glycolytic: 1,970 microm(2)) and equatorial (fast glycolytic: 3,535 microm(2), fast oxidative glycolytic: 1,714 microm(2)) levels. The differences along and across the muscle show the need to design a standardised procedure for sample location when performing comparative studies of morphofunctional adaptive changes in skeletal muscle. A significant difference between individuals (animals) in all parameters was evident and should be taken into consideration when analysing the variability: the factor 'animal' should be considered in multiway ANOVAs, especially when low sample sizes are used.

Capillarity and fibre types in locomotory muscles of wild yellow-legged gulls (Larus cachinnans).

This study analyzes the capillarity and fibre-type distribution of six locomotory muscles of gulls. The morphological basis and the oxygen supply characteristics of the skeletal muscle of a species with a marked pattern of gliding flight are established, thus contributing to a better understanding of the physiology of a kind of flight with low energetic requirements. The four wing muscles studied (scapulotriceps, pectoralis, scapulohumeralis, and extensor metacarpi) exhibited higher percentages of fast oxidative glycolytic fibres (>70%) and lower percentages of slow oxidative fibres (<16%) than the muscles involved in nonflight locomotion (gastrocnemius and iliotibialis). Capillary densities ranged from 816 to 1,233 capillaries mm(-2), having the highest value in the pectoralis. In this muscle, the fast oxidative glycolytic fibres had moderate staining for succinate dehydrogenase and relatively large fibre sizes, as deduced from the low fibre densities (589-665 fibres mm(-2)). All these findings are seen as an adaptive response for gliding, when the wing is held outstretched by isometric contractions. The leg muscles studied included a considerable population of slow oxidative fibres (>14% in many regions), which suggests that they are adapted to postural activities. Regional variations in the relative distributions of fibre types in muscle gastrocnemius may reflect different functional demands placed on this muscle during terrestrial and aquatic locomotion. The predominance of oxidative fibres and capillary densities under 1,000 capillaries mm(-2) in leg muscles is probably a consequence of an adaptation for slow swimming and maintenance of the posture on land rather than for other locomotory capabilities, such as endurance or sprint activities.

Comparative skeletal muscle fibre morphometry among wild birds with different locomotor behaviour.

Six muscles of the mallard duck (Anas platyrhynchos), the common coot (Fulica atra) and the yellow-legged gull (Larus cachinnans) were analysed morphometrically, with special emphasis on their functional implications and physiological needs. Oxidative fibres always had significantly smaller size than anaerobic fibres, although no differences in the number of capillaries per fibre were found. This resulted in greater capillary counts per unit of fibre area and perimeter in oxidative than anaerobic fibres, which indicates that the greater demand for oxygen supply may be achieved by decreasing the size of the muscle fibre rather than by increasing the number of associated capillaries. Fast oxidative fibres of the pectoralis and the triceps of the gull had greater sizes than the fast oxidative fibres of the mallard and the coot, which correlates with the difference in energetic demands between flapping and gliding flight. Greater fibre cross-sectional areas and perimeters seem suited to afford the long-lasting activity with low metabolic demands required during gliding. By contrast, mallards and coots attain a high oxidative metabolism, during sustained flapping flight, by reducing fibre size at the expense of a diminished ability for force generation. Between-species comparisons of the hindlimb muscles only yielded differences for the anaerobic fibres of the gastrocnemius, as an important adaptive response to force generation during burst locomotion. The need to manage sustained swimming abilities effectively may result in similar FOG fibre morphometry of the hindlimb muscles studied, indicating that a compromise between the oxygen flux to the muscle cell and the development of power is highly optimised in oxidative fibres of the bird species studied.

Capillarity and fiber types in locomotory muscles of wild common coots, Fulica atra.

Six locomotory muscles of wild common coots, Fulica atra, were analyzed histochemically. Capillarity and fiber-type distributions were correlated to the functional implications and physiological needs of each muscle. Leg muscles exhibit three unevenly distributed fiber types, a pattern that reflects the great variety of terrestrial and aquatic locomotory performances that coots are able to develop. Aerobic zones are presumably recruited during steady swimming and diving, while regions with anaerobic characteristics may be used for bursts of activity such as sprint swimming or during take off, when coots run along the water's surface. Fiber types and capillarization in wing muscles have a marked oxidative trend. High wing beat frequencies, short and broad wings, and the long distance migrations that these birds perform indicate that the presence of high numbers of oxidative fibers and the well developed capillary supply are needed for enhanced oxygen uptake. The pectoralis muscle, except in its deep part, has exclusively fast oxidative fibers with a very high staining intensity for succinate dehydrogenase assay as compared to the same fiber type of other muscles. Its predominant role in flapping flight justifies these characteristics that are typical of fibers with high aerobic metabolism. The deep part of the pectoralis muscle presents a low proportion of an unusual slow anaerobic fiber type. These fibers could play a role during feeding dives when the bird presses the air out of the feathers by tightening the wings against the body. A linear relationship between capillary and fiber densities in all coot muscles studied reflects an adjustment between fiber diameter and vascularization in order to obtain the oxygen for mitochondrial supply. This strategy seems a suitable way to cope with the rigid aerobic constraints that flying and diving impose upon the coot's physiology. J. Morphol. 237:147-164, 1998. © 1998 Wiley-Liss, Inc.

Innervation distribution pattern, nerve ending structure, and fiber types in pigeon skeletal muscle.

Four fiber types have been characterized in different pigeon skeletal muscles according to their innervation pattern (nerve ending structure and innervation distribution) and histochemical properties (SDH and m-ATPase activities). All fast fibers, types IIA and IIB, present aggregated distribution of their nerve endings with "en plaque" structures and very low innervation frequencies. The two kinds of slow fibers recognized are multiple innervated and present higher innervation frequencies. However, type I fibers have nerve terminals in small knobs with uniform localization, whereas type III fibers present "en grappe" nerve endings, which tend to be randomly distributed. Fiber type composition of skeletal muscles has been found closely related to their biomechanical function. Fast fibers are predominant in muscles with an active role in locomotive movements, whereas slow fibers are mainly or exclusively located in postural muscles.

A combined myosin ATPase and acetylcholinesterase histochemical method for the demonstration of fibre types and their innervation pattern in skeletal muscle.

An improved, combined staining method for myofibrillar ATPase (m-ATPase) and for acetylcholinesterase activity is described. This method allows the observations, on the same slide, of the classical histochemical m-ATPase profile following the Brooke and Kaiser technique and the neuromuscular junction morphology. Thus the pattern of innervation, nerve ending structure and number of nerve endings along the fibres is shown simultaneously for the basic differentiation between slow and fast fibres. The use of acidic and alkaline preincubation allows better visualization of endplate morphology and avoids the masking effect of a positive m-ATPase reaction. The technique has been validated on skeletal muscles from avian and mammalian species.

A histochemical ATPase method for the demonstration of the muscle capillary network.

A histochemical method for demonstration of the capillaries in skeletal muscle of birds is proposed. The present method, which is a modification of a previously reported myosin ATPase technique used for simultaneous staining of capillaries and fiber types, provides an accurate count of capillaries associated with different fiber types in avian skeletal muscles. We have applied the original and the modified method to serial adjacent sections of certain skeletal muscles and our results show that after the application of the original technique: (a) in muscles having dark Type II fibers, these fibers produce a masking effect on their adjacent capillaries; (b) a consistent and significant undercounting in capillary densities can be seen even in muscles having no dark Type II fibers; and (c) the staining quality and capillary count are substantially improved with the use of the modified method. We attribute the better results obtained with our modification to differences in thermosensitivity of ATPase activity from the capillary endothelium and of the myofibers. A mathematical treatment is therefore proposed to correct the values of capillary count obtained with the original method.

Skeletal muscle capillarization and fiber types in urban and homing pigeons (Columba livia).

1. Fiber types, capillary supply and other morphometrical parameters were analysed in pectoral, gastrocnemius and pronator muscles of homing and urban pigeons. 2. The two kinds of birds were analysed before and after a restrainment period of at least 5 months. 3. Only slight differences in fiber type frequencies were noted between urban and homing pigeons in control conditions. 4. The effect of restrainment on the different parameters studied was unclear in gastrocnemius and pronator muscles and negligible in M. pectoralis. 5. Mean diffusion distances for oxygen from capillaries were smaller in oxidative fibers; also, vascularization indexes were higher for these fiber types. 6. The contribution of each fiber type to total sectional ara of muscle remains stable in spite of fiber type frequencies heterogeneity.