John W. Hermanson

Myosin isoforms and muscle fiber characteristics in equine gluteus medius muscle

To date, four different myosin heavy chain (MyHC) isoforms have been identified in adult skeletal muscle of a number of species: types I, IIa, IIx or IId, and IIb. The aim of this study was to investigate the distribution of various MyHC isoforms in the equine gluteus medius and gluteus profundus muscles in relation with several morphometric variables of muscle fibers.

Terrestrial locomotion of the New Zealand short-tailed bat Mystacina tuberculata and the common vampire bat Desmodus rotundus.

SUMMARY Bats (Chiroptera) are generally awkward crawlers, but the common vampire bat (Desmodus rotundus) and the New Zealand short-tailed bat (Mystacina tuberculata) have independently evolved the ability to manoeuvre well on the ground. In this study we describe the kinematics of locomotion in both species, and the kinetics of locomotion in M. tuberculata. We sought to determine whether these bats move terrestrially the way other quadrupeds do, or whether they possess altogether different patterns of movement on the ground than are observed in quadrupeds that do not fly. Using high-speed video analyses of bats moving on a treadmill, we observed that both species possess symmetrical lateral-sequence gaits similar to the kinematically defined walks of a broad range of tetrapods. At high speeds, D. rotundus use an asymmetrical bounding gait that appears to converge on the bounding gaits of small terrestrial mammals, but with the roles of the forelimbs and hindlimbs reversed. This gait was not performed by M. tuberculata. Many animals that possess a single kinematic gait shift with increasing speed from a kinetic walk (where kinetic and potential energy of the centre of mass oscillate out of phase from each other) to a kinetic run (where they oscillate in phase). To determine whether the single kinematic gait of M. tuberculata meets the kinetic definition of a walk, a run, or a gait that functions as a walk at low speed and a run at high speed, we used force plates and high-speed video recordings to characterize the energetics of the centre of mass in that species. Although oscillations in kinetic and potential energy were of similar magnitudes, M. tuberculata did not use pendulum-like exchanges of energy between them to the extent that many other quadrupedal animals do, and did not transition from a kinetic walk to kinetic run with increasing speed. The gait of M. tuberculata is kinematically a walk, but kinetically run-like at all speeds.

Contractile behavior of the forelimb digital flexors during steady-state locomotion in horses (Equus caballus): An initial test of muscle architectural hypotheses about in vivo function

The forelimb digital flexors of the horse display remarkable diversity in muscle architecture despite each muscle-tendon unit having a similar mechanical advantage across the fetlock joint. We focus on two distinct muscles of the digital flexor system: short compartment deep digital flexor (DDF(sc)) and the superficial digital flexor (SDF). The objectives were to investigate force-length behavior and work performance of these two muscles in vivo during locomotion, and to determine how muscle architecture contributes to in vivo function in this system. We directly recorded muscle force (via tendon strain gauges) and muscle fascicle length (via sonomicrometry crystals) as horses walked (1.7 m s(-1)), trotted (4.1 m s(-1)) and cantered (7.0 m s(-1)) on a motorized treadmill. Over the range of gaits and speeds, DDF(sc) fascicles shortened while producing relatively low force, generating modest positive net work. In contrast, SDF fascicles initially shortened, then lengthened while producing high force, resulting in substantial negative net work. These findings suggest the long fibered, unipennate DDF(sc) supplements mechanical work during running, whereas the short fibered, multipennate SDF is specialized for economical high force and enhanced elastic energy storage. Apparent in vivo functions match well with the distinct architectural features of each muscle.

Architectural and Histochemical Analysis of the Biceps brachii Muscle of the Horse

The biceps brachii of horses is a complex muscle subdivided into two heads which may subserve distinct functions. The lateral head contains a large percentage of type I myofibers. This region is largely composed of short fibers (5-7 mm long) arranged in a pinnate fashion and heavily invested with connective tissue. The medial head contains fewer type I fibers and is composed of relatively longer myofibers (15-20 mm long), also arranged in a pinnate fashion but less heavily invested with connective tissue. It is hypothesized that the lateral muscle head of biceps brachii contributes to the postural role of the muscle in the forelimb passive stay apparatus. The medial head, with its longer fibers and generally fast fiber population may be most important during dynamic activity such as walking, trotting and running.

Role of the hypoglossal nerve in equine nasopharyngeal stability

The equine upper airway is highly adapted to provide the extremely high oxygen demand associated with strenuous aerobic exercise in this species. The tongue musculature, innervated by the hypoglossal nerve, plays an important role in airway stability in humans who also have a highly adapted upper airway to allow speech. The role of the hypoglossal nerve in stabilizing the equine upper airway has not been established. Isolated tongues from eight mature horses were dissected to determine the distal anatomy and branching of the equine hypoglossal nerve. Using this information, a peripheral nerve location technique was used to perform bilateral block of the common trunk of the hypoglossal nerve in 10 horses. Each horse was subjected to two trials with bilateral hypoglossal nerve block and two control trials (unblocked). Upper airway stability at exercise was determined using videoendoscopy and measurement of tracheal and pharyngeal pressure. Three main nerve branches were identified, medial and lateral branches and a discrete branch that innervated the geniohyoid muscle alone. Bilateral hypoglossal block induced nasopharyngeal instability in 10/19 trials, and none of the control trials (0/18) resulted in instability (P<0.001). Mean treadmill speed (+/-SD) at the onset of instability was 10.8+/-2.5 m/s. Following its onset, nasopharyngeal instability persisted until the end of the treadmill test. This instability, induced by hypoglossal nerve block, produced an expiratory obstruction similar to that seen in a naturally occurring equine disease (dorsal displacement of the soft palate, DDSP) with reduced inspiratory and expiratory pharyngeal pressure and increased expiratory tracheal pressure. These data suggest that stability of the equine upper airway at exercise may be mediated through the hypoglossal nerve. Naturally occurring DDSP in the horse shares a number of anatomic similarities with obstructive sleep apnea. Study of species with extreme respiratory adaptation, such as the horse, may provide insight into respiratory functioning in humans.

Superficial digital flexor tendon lesions in racehorses as a sequela to muscle fatigue : A preliminary study

REASON FOR PERFORMING STUDY Racing and training related lesions of the forelimb superficial digital flexor tendon are a common career ending injury to racehorses but aetiology and/or predisposing causes of the injury are not completely understood. OBJECTIVES Although the injury takes place within the tendon, the lesion must be considered within the context of the function of the complete suspensory system of the distal limb, including the associated muscles. METHODS Both muscle and tendon function were investigated in vivo using implanted strain gauges in 3 Thoroughbred horses walking, trotting and cantering on a motorised treadmill. These data were combined with assessments of muscle architecture and fibre composition to arrive at an overview of the contribution of each muscle-tendon unit during locomotion. RESULTS The superficial digital flexor muscle has fatigue-resistant and high force production properties that allow its tendon to store and return elastic energy, predominantly at the trot. As running speed increases, deep digital flexor tendon force increases and it stabilises hyperextension of the fetlock, thus reinforcing the superficial digital flexor in limb load support. The deep digital flexor muscle has fast contracting properties that render it susceptible to fatigue. CONCLUSION Based on these measurements and supporting evidence from the literature, it is proposed that overloading of the superficial digital flexor tendon results from fatigue of the synergistic, faster contracting deep digital flexor muscle. POTENTIAL RELEVANCE Future research investigating distal limb system function as a whole should help refine clinical diagnostic procedures and exercise training approaches that will lead to more effective prevention and treatment of digital flexor tendon injuries in equine athletes.

Forelimb versus Hindlimb Skeletal Development in the Big Brown Bat, Eptesicus fuscus: Functional Divergence Is Reflected in Chondrocytic Performance in Autopodial Growth Plates

The morphology of the chiropteran forelimb demonstrates musculoskeletal specializations for powered flight essentially unique among mammals, including extreme elongation of the distal skeletal elements. Recent studies have focused primarily on the relative timing and levels of gene expression during early stages of endochondral ossification in the chiropteran embryo for clues to the molecular basis of the evolutionary origins of flight in these species. The goal of the current study was to examine how elongation of skeletal elements of the forelimb autopod is achieved through a combination of cellular proliferation, cellular enlargement and matrix synthesis during a period of rapid postnatal growth in Eptesicus fuscus. Quantitative analyses were done of multiple performance parameters of growth plate chondrocytes during all phases of the differentiation cascade. Fourteen autopodial growth plates from the forelimb and hindlimb of one individual, as well as the proximal tibial growth plate, were collected and analyzed. Significant differences were seen in all performance parameters examined. Particularly striking were the differences between growth plates of the manus and pes in the size of the pool of chondrocytes in all cellular zones and rates of turnover of terminal cells. The magnitude of hypertrophy of chondrocytes in growth plates of the manus in E. fuscus far exceeded what has been reported previously in any species, even in rapidly elongating rodent long bones. Volume changes approaching ×70 and height changes of 50–60 µm/cell (paralleling the direction of growth) occurred after proliferation in the most rapidly growing growth plates. The data demonstrate that final differences in lengths of homologous skeletal elements in the autopod of the forelimb and hindlimb of this species result not just from an initiating factor early in development, but from continued quantitative differences in chondrocytic performance during postnatal bone elongation as measured by multiple kinetic-based parameters.

Testing the hindlimb-strength hypothesis: non-aerial locomotion by Chiroptera is not constrained by the dimensions of the femur or tibia

SUMMARY In the evolution of flight bats appear to have suffered a trade-off; they have become poor crawlers relative to terrestrial mammals. Capable walking does occur in a few disparate taxa, including the vampire bats, but the vast majority of bats are able only to shuffle awkwardly along the ground, and the morphological bases of differences in crawling ability are not currently understood. One widely cited hypothesis suggests that the femora of most bats are too weak to withstand the compressive forces that occur during terrestrial locomotion, and that the vampire bats can walk because they possess more robust hindlimb skeletons. We tested a prediction of the hindlimb-strength hypothesis: that during locomotion, the forces produced by the hindlimbs of vampire bats should be larger than those produced by the legs of poorly crawling bats. Using force plates we compared the hindlimb forces produced by two species of vampire bats that walk well, Desmodus rotundus (N=8) and Diaemus youngi (N=2), to the hindlimb forces produced during over-ground shuffling by a similarly sized bat that is a poor walker (Pteronotus parnellii; N=6). Peak hindlimb forces produced by P. parnellii were larger (ANOVA; P<0.05; N=65) and more variable (93.5±36.6% body weight, mean ± s.d.) than those of D. rotundus (69.3±8.1%) or D. youngi (75.0±6.2%). Interestingly, the vertical components of peak force were equivalent among species (P>0.6), indicating similar roles for support of body weight by the hindlimbs in the three species. We also used a simple engineering model of bending stress to evaluate the support capabilities of the hindlimb skeleton from the dimensions of 113 museum specimens in 50 species. We found that the hindlimb bones of vampires are not built to withstand larger forces than those of species that crawl poorly. Our results show that the legs of poorly crawling bats should be able to withstand the forces produced during coordinated crawling of the type used by the agile vampires, and this indicates that some mechanism other than hindlimb bone thickness, such as myology of the pectoral girdle, limits the ability of most bats to crawl.

Elbow extensor muscles of the horse: postural and dynamic implications.

Based on histochemical and immunohistochemical evidence, horse elbow extensor muscles are composed of two morphologically distinct muscle groups. The long and lateral heads of the triceps brachii are large, predominantly type II (presumed fast) muscles. The long and lateral heads of the triceps together account for 96% of the weight of the elbow extensors (long head of triceps is 81%). The long and lateral heads contain three histochemical fiber types: types I, IIa and IIb. Type I muscle fibers account for approximately 18 and 27% of the fibers in the long and lateral heads of the triceps, respectively. In the lateral head, type IIa and IIb fibers account equally for the remaining 70%, while in the long head of the triceps type IIb fibers predominate (50%) over type IIa fibers (32%). In contrast, the much smaller medial head of the triceps (2% of triceps mass) and the anconeus (2% of mass) contain almost exclusively type I muscle fibers. It is hypothesized that the medial head and anconeus, with their slow fibers, contribute to the postural maintenance of the forelimb by preventing flexion at the elbow joint during passive stance. The larger long and lateral heads, with their generally fast fiber populations, are most likely important during dynamic activity.

Neuroanatomy of the equine dorsal cricoarytenoid muscle: Surgical implications

REASON FOR PERFORMING STUDY Studies are required to define more accurately and completely the neuroanatomy of the equine dorsal cricoarytenoid muscle as a prerequisite for developing a neuroprosthesis for recurrent laryngeal neuropathy. OBJECTIVES To describe the anatomy, innervation, fibre types and function of the equine dorsal cricoarytenoid muscle. METHODS Thirty-one larynges were collected at necropsy from horses with no history of upper airway disease and 25 subjected to gross dissection. Thereafter, the following preparations were made on a subset of larynges: histochemical staining (n = 5), Sihlers and acetylcholinesterase staining for motor endplates (n = 2). An additional 6 larynges were collected and used for a muscle stimulation study. RESULTS Two neuromuscular compartments (NMC), each innervated by a primary nerve branch of the recurrent laryngeal nerve, were identified in all larynges. Stimulation of the lateral NMC produced more lateral displacement of the arytenoid cartilage than the medial NMC (P<0.05). The medial NMC tended to rotate the arytenoid cartilage dorsally. Motor endplates were identified at the junction of the middle and caudal thirds of each NMC. If fibre type grouping was present it was always present in both NMCs. CONCLUSIONS The equine dorsal cricoarytenoid muscle has 2 distinct muscle NMCs with discrete innervation and lines of action. The lateral NMC appears to have a larger role in increasing cross-sectional area of the rima glottidis. POTENTIAL RELEVANCE This information should assist in planning surgical reinnervation procedures and development of a neuroprosthesis for recurrent laryngeal neuropathy.