Immunohistochemistry of kangaroo rat hindlimb muscles.

Kangaroo rats (Dipodomys spp.) use specialized bipedal hopping like that of kangaroos. In contrast to kangaroos that have elastic tendons capable of storing energy, kangaroo rats have inelastic tendons that are unable to store large amounts of energy. Thus, the musculature of the ankle joint provides the greatest power contribution to kangaroo rat hopping. Skeletal muscle can be characterized by several fiber types, including slow twitch (Type I) and fast twitch (Type II) fibers. Fast fibers are found in higher concentration in muscles that perform quick, dynamic movements, whereas slow fibers are found in higher proportion in muscles that perform slow, endurant movements. Using fiber type specific antibodies, we identified four pure (Types I, IIA, IIB, and IIX) and two hybrid (Types I/IIA and IIA/IIX) fiber types in six hindlimb muscles from three kangaroo rats (Dipodomys merriami) to investigate the relationship between fiber composition and hindlimb muscle function. Hindlimb muscles (except soleus) were dominated by Type IIB fibers, which were largest in cross-sectional area, and are known to be best suited for rapid and explosive movements. Oxidative Type IIA and Type IIX fibers were found at moderate concentrations and likely function in maintaining continual saltatory locomotion. Thus, kangaroo rats can use these two fiber type populations as "gears" for both endurant and explosive behaviors.

The anatomy and histochemistry of flight hindlimb posture in birds. II. The flexed hindlimb posture of perching birds.

During flight, birds employ one of two hindlimb postures. Perching birds utilize a flexed posture with their folded legs tucked beneath the body, whereas shorebirds and raptors use an extended posture with straightened legs trailing behind the body. Maintenance of either posture during flight requires the hindlimbs to hold their position for prolonged periods. Slow contracting fibers are known for their fatigue-resistant properties and are often found in high percentages in muscles utilized for postural actions. Given the similar actions required of the hip and knee flexors used during flight, we hypothesized that the equivalent postural muscles of perching birds (flexed posture) would contain similar percentages of slow fibers as shorebirds (extended posture). We investigated the anatomy and fiber type composition of seven hindlimb muscles in yellow-headed and red-winged blackbirds and revealed that they possess a smaller percentage of slow fibers than we found previously in the same muscles of American avocets and black-necked stilts. The comparably smaller body size of yellow-headed and red-winged blackbirds could mitigate the need for more slow fibers. In addition, the biomechanical placement of the weight force in the flexed posture may require less muscle force for postural support during flight and, therefore, fewer slow fibers.

Anatomy and histochemistry of spread-wing posture inbirds. 4. Eagles soar with fast, not slow muscle fibres.

Slow fibers are typically characterized as functioning in avian postural behaviors such as soaring flight, and are described for a number of elite soarers such as vultures, pelicans and albatrosses. Golden Eagles and Bald Eagles also display soaring behavior and we examined their flight muscles for the presence of slow fibers. Surprisingly, eagles lack a deep layer to the pectoralis found in other soaring species. Additionally, the pectoralis as well as other shoulder muscles had few to no slow muscle fibers. The lack of functionally meaningful numbers of slow muscle fibers in eagle flight muscles indicates that they must rely on fast fibers for posture; these can function in that role due to their high aerobic capacity and also perhaps a "tuning" of muscle contraction frequency to function more efficiently at isometric contractions.

Anatomy and histochemistry of hindlimb flight posture in birds. I. The extended hindlimb posture of shorebirds.

Birds utilize one of two hindlimb postures during flight: an extended posture (with the hip and knee joints flexed, while the ankle joint is extended caudally) or a flexed posture (with the hip, knee, and ankle joints flexed beneath the body). American Avocets (Recurvirostra americana) and Black-necked Stilts (Himantopus mexicanus) extend their legs caudally during flight and support them for extended periods. Slow tonic and slow twitch muscle fibers are typically found in muscles functioning in postural support due to the fatigue resistance of these fibers. We hypothesized that a set of small muscles composed of high percentages of slow fibers and thus dedicated to postural support would function in securing the legs in the extended posture during flight. This study examined the anatomy and histochemical profile of eleven hindlimb muscles to gain insight into their functional roles during flight. Contrary to our hypothesis, all muscles possessed both fast twitch and slow twitch or slow tonic fibers. We believe this finding is due to the versatility of dynamic and postural functions the leg muscles must facilitate, including standing, walking, running, swimming, and hindlimb support during flight. Whether birds use an extended or flexed hindlimb flight posture may be related to the aerodynamic effect of leg position or may reflect evolutionary history.

Horse soleus muscle: postural sensor or vestigial structure?

The soleus muscle of horses is rather diminutive with respect to the overall size of adjacent synergist muscles in the hind limb of the horse. Whether or not such a muscle might be vestigial or may be providing some essential function has not been determined. We have studied the horse's soleus muscle using histochemical (ATPase), immunocytochemical (myosin isoform identification), and SDS-PAGE analysis to demonstrate that it is largely composed of 100% type I, presumed slow-twitch fibers. Only one soleus muscle studied (out of 13 adult horses) contained any type II muscle fibers. Given this consistent high percentage of slow-oxidative fibers, we hypothesized that the soleus muscle could have a significant role in proprioceptive function, essentially functioning as a proprioceptive organ instead of a significant force-generating muscle during locomotion. We tested this by examining three whole soleus muscles and assessing their muscle spindle content, which proved to have a spindle index of about 12. This value provided equivocal support for the hypothesis since it did not approach values reported for other mammalian proprioceptive muscles that were approximately 40-50 spindles per gram of muscle mass. Other parameters, such as motoneuron number and muscle unit size, may be useful in understanding these data.

Anatomy and histochemistry of spread-wing posture in birds. 3. Immunohistochemistry of flight muscles and the "shoulder lock" in albatrosses.

As a postural behavior, gliding and soaring flight in birds requires less energy than flapping flight. Slow tonic and slow twitch muscle fibers are specialized for sustained contraction with high fatigue resistance and are typically found in muscles associated with posture. Albatrosses are the elite of avian gliders; as such, we wanted to learn how their musculoskeletal system enables them to maintain spread-wing posture for prolonged gliding bouts. We used dissection and immunohistochemistry to evaluate muscle function for gliding flight in Laysan and Black-footed albatrosses. Albatrosses possess a locking mechanism at the shoulder composed of a tendinous sheet that extends from origin to insertion throughout the length of the deep layer of the pectoralis muscle. This fascial "strut" passively maintains horizontal wing orientation during gliding and soaring flight. A number of muscles, which likely facilitate gliding posture, are composed exclusively of slow fibers. These include Mm. coracobrachialis cranialis, extensor metacarpi radialis dorsalis, and deep pectoralis. In addition, a number of other muscles, including triceps scapularis, triceps humeralis, supracoracoideus, and extensor metacarpi radialis ventralis, were found to have populations of slow fibers. We believe that this extensive suite of uniformly slow muscles is associated with sustained gliding and is unique to birds that glide and soar for extended periods. These findings suggest that albatrosses utilize a combination of slow muscle fibers and a rigid limiting tendon for maintaining a prolonged, gliding posture.

Anatomy and histochemistry of spread-wing posture in birds. I. Wing drying posture in the double-crested cormorant, Phalacrocorax auritus.

Spread-wing postures of birds often have been studied with respect to the function of behavior, but ignored with regard to the mechanism by which the birds accomplish posture. The double-crested cormorant, Phalacrocorax auritus, was used as a model for this study of spread-wing posture. Those muscles capable of positioning and maintaining the wing in extension and protraction were assayed histochemically for the presence of slow (postural) muscle fibers. Within the forelimb of Phalacrocorax, Mm. coracobrachialis cranialis, pectoralis thoracicus (cranial portion), deltoideus minor, triceps scapularis, and extensor metacarpi radialis pars dorsalis and ventralis were found to contain populations of slow-twitch or slow-tonic muscle fibers. These slow fibers in the above muscles are considered to function during spread-wing posture in this species. J Morphol 233:67-76, 1997. © 1997 Wiley-Liss, Inc.

Gliding flight in the American Kestrel (Falco sparverius): An electromyographic study.

Electromyographic (EMG) activity was studied in American Kestrels (Falco sparverius) gliding in a windtunnel tilted to 8 degrees below the horizontal. Muscle activity was observed in Mm. biceps brachii, triceps humeralis, supracoracoideus, and pectoralis, and was absent in M. deltoideus major and M. thoracobrachialis (region of M. pectoralis). These active muscles are believed to function in holding the wing protracted and extended during gliding flight. Quantification of the EMG signals showed a lower level of activity during gliding than during flapping flight, supporting the idea that gliding is a metabolically less expensive form of locomotion than flapping flight. Comparison with the pectoralis musculature of specialized gliding and soaring birds suggests that the deep layer of the pectoralis is indeed used during gliding flight and that the slow tonic fibers found in soaring birds such as vultures represents a specialization for endurant gliding. It is hypothesized that these slow fibers should be present in the wing muscles that these birds use for wing protraction and extension, in addition to the deep layer of the pectoralis. © 1993 Wiley-Liss, Inc.