Anatomic and neuromuscular characterisation of the equine cricothyroid muscle.

REASONS FOR PERFORMING STUDY: As part of investigation into laryngeal stability and reanimation using functional electrical stimulation, the cricothyroid muscle might be utilised to increase laryngeal cross-sectional area in horses with recurrent laryngeal neuropathy. For optimal electrode placement and muscle recruitment, the neuroanatomy and excitability of the equine cricothyroid muscle needs to be defined. OBJECTIVES: To describe the anatomy, innervation and function of the equine cricothyroid muscle and its contribution to laryngeal diameter. METHODS: Seventeen equine larynges were collected at necropsy and 12 were grossly dissected. Seven larynges (five grade 1, two grade 4) were prepared for immunohistochemistry following gross dissection and 5 larynges were prepared for special staining: acetylcholinesterase staining of motor endplates (n = 3) and Sihler's staining (n = 2). Three larynges were stimulated following in situ cadaver dissection and 2 larynges were removed and stimulated ex vivo. RESULTS: Three neuromuscular compartments, each innervated by a primary nerve branch of the external branch of the cranial laryngeal nerve, were identified in all larynges. Stimulation of each neuromuscular compartment resulted in ventral displacement of the thyroid cartilage with respect to the cricoid cartilage, thereby increasing dorsoventral height of the rima glottis. CONCLUSIONS: The equine cricothyroid muscle has 3 distinct neuromuscular compartments with discrete innervation, fibre type distribution and muscle fibre sizes. All neuromuscular compartments tense the vocal cords by increasing dorsoventral height of the rima glottis through ventral displacement of the thyroid cartilage with respect to the cricoid cartilage. POTENTIAL RELEVANCE: Simultaneous functional electrical stimulation of the cricothyroid and dorsal cricoarytenoid muscles may enhance laryngeal cross-sectional area in horses with recurrent laryngeal neuropathy.

Contractile properties of muscle fibers from the deep and superficial digital flexors of horses.

Equine digital flexor muscles have independent tendons but a nearly identical mechanical relationship to the main joint they act upon. Yet these muscles have remarkable diversity in architecture, ranging from long, unipennate fibers ("short" compartment of DDF) to very short, multipennate fibers (SDF). To investigate the functional relevance of the form of the digital flexor muscles, fiber contractile properties were analyzed in the context of architecture differences and in vivo function during locomotion. Myosin heavy chain (MHC) isoform fiber type was studied, and in vitro motility assays were used to measure actin filament sliding velocity (V(f)). Skinned fiber contractile properties [isometric tension (P(0)/CSA), velocity of unloaded shortening (V(US)), and force-Ca(2+) relationships] at both 10 and 30°C were characterized. Contractile properties were correlated with MHC isoform and their respective V(f). The DDF contained a higher percentage of MHC-2A fibers with myosin (heavy meromyosin) and V(f) that was twofold faster than SDF. At 30°C, P(0)/CSA was higher for DDF (103.5 ± 8.75 mN/mm(2)) than SDF fibers (81.8 ± 7.71 mN/mm(2)). Similarly, V(US) (pCa 5, 30°C) was faster for DDF (2.43 ± 0.53 FL/s) than SDF fibers (1.20 ± 0.22 FL/s). Active isometric tension increased with increasing Ca(2+) concentration, with maximal Ca(2+) activation at pCa 5 at each temperature in fibers from each muscle. In general, the collective properties of DDF and SDF were consistent with fiber MHC isoform composition, muscle architecture, and the respective functional roles of the two muscles in locomotion.

Considerations for pacing of the cricoarytenoid dorsalis muscle by neuroprosthesis in horses.

REASONS FOR PERFORMING STUDY: The success rate of prosthetic laryngoplasty is limited and may be associated with significant sequelae. Nerve muscle pedicle transplantation has been attempted but requires a year before function is restored. OBJECTIVE: To determine the optimal parameters for functional electrical stimulation of the recurrent laryngeal nerve in horses. METHODS: An experimental in vivo study was performed on 7 mature horses (2-21 years). A nerve cuff was placed on the distal end of the common trunk of the recurrent laryngeal nerve (RLN). In 6 horses the ipsilateral adductor branch of RLN was also transected. The electrodes were connected to programmable internal stimulator. Stimulation was performed using cathodic phase and then biphasic pulses at 24 Hz with a 0.427 ms pulse duration. Stimulation-response experiments were performed at monthly intervals, from one week following implantation. The study continued until unit failure or the end of project (12 months). Two of the horses were stimulated continuously for 60 min to assess onset of fatigue. RESULTS: Excellent arytenoid cartilage abduction (mean arytenoid angle of 52.7 degrees, range 48.5-56.2 degrees) was obtained in 6 horses (laryngeal grades I or II (n = 3) and III (n = 2). Poor abduction was obtained in grade IV horses (n = 2). Arytenoid abduction was maintained for up to a year in one horse. Technical implant failure resulted in loss of abduction in 6 horses at one week to 11 months post operatively. Mean tissue impedance was 1.06 kOhm (range 0.64-1.67 kOhm) at one week, twice this value at 2 months (mean 2.32, range 1.11-3.75 kOhm) and was stable thereafter. Maximal abduction was achieved at a stimulation range of 0.65-7.2 mA. No electrical leakage was observed. Constant stimulation of the recurrent laryngeal nerve for 60 min led to full abduction without evidence of muscle fatigue. CONCLUSIONS: Functional electrical stimulation of the recurrent laryngeal nerve leading to full arytenoid abduction can be achieved. The minimal stimulation amplitude for maximal abduction angle is slightly higher than those for man and dogs. CLINICAL RELEVANCE: This treatment modality could eventually be applicable to horses with recurrent laryngeal neuropathy.

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.

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), Sihler's 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.

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.

Morphology, histochemistry, and function of epaxial cervical musculature in the horse (Equus caballus).

The semispinalis capitis and splenius muscles of the horse were analyzed for gross morphology, microarchitecture, fiber length, and fiber type. Although these two muscles are similar in size and anatomical position, they are very different from one another in structural design and histochemistry, implying diverse functional roles in the animal's behavior. The histochemical staining profile was limited to two fiber types: slow oxidative and fast glycolytic. The splenius muscle has simple architecture, long fibers, and a 60/40 ratio of SO to FG cross-sectional area. The semispinalis capitis has complex architecture with short-fibered, concentric compartments dorsal to its central tendon and longer-fibered compartments ventrally. The entire dorsal region has an increasing gradient of slow oxidative fiber percentage from caudal to cranial (58-71% SO). In contrast, the ventral region has a decreasing gradient of slow oxidative fibers from caudal to cranial (48-67% FG). These patterns can be interpreted within the context of the cervical musculature during locomotion and posture to indicate the functional advantages of this organization.

Segmental in vivo vertebral kinematics at the walk, trot and canter: a preliminary study.

Understanding the pathophysiology of equine back problems, for clinical evaluation, treatment or injury prevention, requires understanding of the normal 3-dimensional motion characteristics of the vertebral column. Recent studies have investigated regional vertebral kinematics; however, there are no reported measures of direct in vivo segmental vertebral kinematics in exercising horses. Relative movements between 2 adjacent vertebrae were recorded for 3 horses that were clinically sound and did not have a known history of a back problem. A transducer consisting of 2 fixtures and an array of liquid metal strain gauges (LMSGs) was used to measure 3-dimensional segmental vertebral motion. The transducer was attached directly to Steinmann pins implanted in the dorsal spinous processes of adjacent vertebrae in 3 vertebral regions: thoracic (T14 to T16), lumbar (L1 to L3) and lumbosacral (L6 to S2). Rotational displacements between adjacent vertebrae were calculated from the differential outputs of the LMSG array during walk, trot and canter on a treadmill. Peak magnitudes of dorsoventral flexion, lateral bending and axial rotation were recorded continuously for each stride. The largest motion of the 3 instrumented vertebral segments was at the lumbosacral junction. In general, the greatest magnitude of segmental vertebral motion occurred during the canter and the least during the trot. The dynamic and continuous measure of 3-dimensional in vivo segmental vertebral motion provides an important new perspective for evaluating vertebral motion and back problems in horses.

Effects of birth weight and postnatal nutrition on neonatal sheep: II. Skeletal muscle growth and development.

This study investigated effects of birth weight and postnatal nutrition on growth and development of skeletal muscles in neonatal lambs. Low (L; mean +/- SD 2.289 +/- .341 kg, n = 28) and high (H; 4.840 +/- .446 kg, n = 20) birth weight male Suffolk x (Finnsheep x Dorset) lambs were individually reared on a liquid diet to grow rapidly (ad libitum fed, ADG 337 g, n = 20) or slowly (ADG 150 g, n = 20) from birth to live weights (LW) up to approximately 20 kg. At birth, weight of semitendinosus (ST) muscle in L lambs was 43% that in H lambs; aggregate weights of ST and seven other dissected muscles were similarly reduced. In ST muscle of L lambs, mass of DNA, RNA, and protein were also significantly reduced to levels 67, 60, and 34%, respectively, of those in H lambs. However, myofiber numbers of ST, tibialis caudalis, or soleus muscles did not differ between the L and H birth weight lambs and did not change during postnatal growth. During postnatal rearing, daily accretion rate of dissected muscle was lower in L than in H lambs. Accretion of muscle per kilogram of gain in empty body weight (EBW) was reduced in the slowly grown L lambs compared with their H counterparts, although the difference was less pronounced between the rapidly grown L and H lambs. Throughout the postnatal growth period, ST muscle of L lambs contained less DNA with a higher protein:DNA ratio at any given muscle weight than that of H lambs. Slowly grown lambs had heavier muscles at any given EBW than rapidly grown lambs. Content of DNA and protein:DNA ratio in ST muscle were unaffected by postnatal nutrition, but RNA content and RNA:DNA were greater and protein:RNA was lower at any given muscle weight in rapidly grown lambs. Results suggest that myofiber number in fetal sheep muscles is established before the presumed, negative effects of inadequate fetal nutrient supply on skeletal muscle growth and development become apparent. However, proliferation of myonuclei may be influenced by fetal nutrition in late pregnancy. Reduced myonuclei number in severely growth-retarded newborn lambs may limit the capacity for postnatal growth of skeletal muscles.

Intrauterine growth retardation is associated with reduced cell cycle activity, but not myofibre number, in ovine fetal muscle.

Cellular development of muscle was studied in sheep fetuses at 85 days of gestation. Large and small fetuses were compared at 100, 115 and 130 days, and an additional group of large 130-day fetuses were studied following 7 days of maternal undernutrition. Myogenesis in the peroneus longus muscle was completed between 100 and 115 days of gestation, and myofibre number did not differ between small and large fetuses. The proportion of myofibre-related nuclei identified as entering S-phase of the cell cycle was 1.7% per hour in 85-day fetuses. In large fetuses, subsequent rates were relatively constant (approximately 1.5% h(-1)), whereas in small fetuses cell cycle activity declined with age from 1.3 to 0.9% h(-1), and was 0.5% h(-1) in 130-day fetuses of restricted ewes. The constant rate of cell cycle activity in large fetuses was associated with an increasing estimated rate of muscle growth (peroneus longus (mg) = 0.831 x 10(0.024 x age [d]), r2 = 0.98), which contrasted with slow and relatively constant muscle accretion in small fetuses (8.4 mg day(-1)), and slower muscle accretion at 130 days in large fetuses from restricted ewes. Differences in DNA and RNA content in the semimembranosus muscle increased with age, large fetuses having 70% more muscle DNA, 108% more muscle RNA and 104% larger muscles than small fetuses at 130 days (all P<0.001). The results demonstrate that myonuclei accumulation, but not myofibre number, is associated with fetal growth in sheep and, therefore, with fetal nutrition during mid to late gestation.

An ultrasound-guided procedure to administer a label of DNA synthesis into fetal sheep.

A novel technique was developed to deliver a bolus dose of a DNA label into the peritoneal cavity of fetal sheep at 85-130 days gestation. Use of markers to identify the site of injection in fetuses from litters up to quadruplets, and immunohistochemistry to detect the DNA label, 5-bromo-2'-deoxyuridine (BrdU), confirmed the procedure was successful in 85% of cases. Duration of the procedure was (mean +/- SD) 44 +/- 16 min, and recovery from anaesthesia was rapid and uneventful in all cases. Fetal weight was estimated with a high degree of accuracy (residual standard deviation (RSD) = 297 g and r2 = 0.93, P<0.001) and the dose of label administered (110 +/- 33 mg BrdU/kg fetal weight) was adequate in all cases. BrdU detected in fetal nuclei following injection into amniotic fluid highlights the need for positive identification of the injection site in timed, short-term studies, and suggests potential to further develop the technique to investigate cellular events in fetal sheep younger than 85 days of gestation. The results demonstrate that the procedure can be used to determine in vivo whether or not nuclei have entered the S-phase of the cell cycle.

Effects of birth weight and postnatal nutrition on neonatal sheep: I. Body growth and composition, and some aspects of energetic efficiency.

We investigated the effects of birth weight and postnatal nutrition on growth characteristics of neonatal lambs. Low- and high-birth-weight male lambs were individually reared on a high-quality liquid diet to grow rapidly (ad libitum access to feed) or slowly (ADG 150 g) to various weights up to 20 kg live weight (LW). Average daily gain tended to be greater in the high- (mean+/-SE 345+/-14 g) than in the low- (329+/-15 g) birth-weight lambs given ad libitum access to feed owing to slower growth by the small newborns during the immediate postpartum period. At birth, on a weight-specific basis, small newborns contained 6.4% less nitrogen and tended to have more ash (8.9%) than the high-birth-weight newborns. Daily rates of fat, ash, and GE accretion were greater, and nitrogen accretion tended to be greater in the rapidly grown large newborns than in their small counterparts. At any given empty body weight (EBW) during rearing, low-birth-weight lambs contained more fat and less ash, resulting in slowly and rapidly grown small newborns containing 39.3 and 42.7 Mcal GE, respectively, at completion of the study (17.5 kg EBW), compared with 34.8 and 40.5 Mcal in their large counterparts. The differences in fatness and energy content between the birth weight categories are attributed to energy requirements for maintenance that were approximately 30% lower, coupled with higher relative intakes in the low-birthweight lambs, during the early postnatal period. At this time, the ability to consume nutrients in excess of lean tissue growth requirements was apparently more pronounced in small than in large newborns, which resulted in lower efficiency of energy utilization for tissue deposition. Furthermore, body composition differences between the slowly and rapidly reared lambs support the notion of a priority of lean tissue over fat when nutrient supply is limited.

Architecture and the division of labor in the extensor carpi radialis muscle of horses.

The extensor carpi radialis muscle of the horse is deceptive at first appearance. It has a fusiform shape similar to other forearm extensor muscles. The fiber arrangement also appears long and relatively parallel. However, it may contain two or more compartments that correlate with differing functional roles. Histochemical and immunocytochemical analysis of proximal and distal regions of the muscle (n = 9) demonstrate that the proximal portion of the muscle is composed of a mean of 13% type I, presumed slow twitch, and 61% type IIb, presumed fast twitch fibers. In contrast, the distal compartment is composed of a mean of about 43% type I and only 22% type IIB fibers. The type I and IIa fibers are all highly aerobic based on nicotinamide dinucleotide tetrazolium reductase reactions. Correlative data regarding the myosin isoforms has been obtained with 4% SDS-PAGE analysis of myosin heavy chain isoforms which demonstrate isoforms migrating at rates similar to rat type I, IIa, and IIx. The latter has been referred to as type IIB/X in a study of the horse's gluteus medius muscle. We propose that the in-series 'compartmentalization' of the muscle, while not conforming strictly to the definitions of neuromuscular compartments, relates to the insertion of the lacertus fibrosus, a distal slip of the biceps brachii, upon the extensor carpi radialis. Earlier studies demonstrated a high proportion of type I fibers in the equine lateral biceps brachii which were thought to stabilize the shoulder during long periods of quiet standing. Because of action imposed on the distal compartment by the biceps brachii, slow and fatigue-resistant functions are part of the limb's passive stay apparatus to effect long-term standing by the horse. Thus, the fatigue-resistant compartments of biceps brachii and extensor carpi radialis may constitute an in-series arrangement of the two muscles. The proximal compartment is suited to provide powerful, more fatigable contractions during locomotion and likely affects stress or strain within the distal postural compartment.

The dynamics of flight-initiating jumps in the common vampire bat Desmodus rotundus.

Desmodus rotundus, the common vampire bat (Phyllostomidae: Desmodontinae), exhibits complex and variable terrestrial movements that include flight-initiating vertical jumps. This ability is unique among bats and is related to their unusual feeding behavior. As a consequence of this behavior, the wing is expected to have design features that allow both powered flight and the generation of violent jumps. In this study, high-speed cine images were synchronized with ground reaction force recordings to evaluate the dynamics of jumping behavior in D. rotundus and to explore the functional characteristics of a wing operating under competing mechanical constraints. The pectoral limbs are responsible for generating upward thrust during the jump. The hindlimbs stabilize and orient the body over the pectoral limbs. The thumbs (pollices) stabilize the pectoral limb and contribute to extending the time over which vertical force is exerted. Peak vertical force can reach 9.5 times body weight in approximately 30 ms. Mean impulse is 0.0580+/-0.007 N s (mean +/- s.d., N=12), which accelerates the animal to a mean take-off velocity of 2.38+/-0.24 m s-1. A model of the muscular activity during jumping is described that accounts for the characteristic force output shown by these animals during flight-initiating jumps.

Myosin isoforms and muscle fiber characteristics in equine gluteus medius muscle.

BACKGROUND: 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. METHODS: Samples from different depths of the gluteus medius muscle (2, 4, 6, and 8 cm) and gluteus profundus muscle of five sedentary horses were examined by MyHC gel electrophoresis, monoclonal antibodies staining against fast, slow and neonatal MyHC isoforms, myosin adenosine triphosphatase (m-ATPase) activity, nicotinamide adenine dinucleotide tetrazolium reductase, alpha-glycerophosphate dehydrogenase, and alpha-amylase-PAS. Data about relative frequencies, sizes, and capillaries of the various histochemical fiber types were collected by morphometry. RESULTS: Three MyHC isoforms were present in the gluteus medius muscle. Two of them comigrated with type I and IIa MyHC isoforms of rat diaphragm (used as a control). The third isoform showed an electrophoretic mobility closer to type IIx than to the IIb MyHC isoform of rat diaphragm. Only two MyHC isoforms (type I and IIa) were detected in the gluteus profundus muscle. In both muscles, type I fibers (high m-ATPase activity at pH 4.5) only reacted with the anti slow-MyHC antibody and both type IIA and IIB fibers (low and moderate m-ATPase activity at pH 4.5, respectively) only reacted with the anti fast-MyHC antibody. No cross-reactivity of fibers positive for both antibodies was found except for the scarce type IIC fibers. Fiber types and capillaries were heterogeneously distributed across the gluteus medius muscle. The deeper regions of this muscle were found to contain a higher percentage of type I fibers, a large number of capillaries and a lower proportion of type IIB fibers compared to the superficial regions of the muscle. The gluteus profundus muscle had more abundant and larger type I fibers than the deepest sampling site of the gluteus medius muscle. CONCLUSIONS: These results show the existence of three different MyHC isoforms in the equine gluteus medius muscle and that fiber types and MyHC isoforms are heterogeneously distributed within this muscle. The distribution of slow-twitch and fast-twitch MyHCs among the fibers determined by immunohistochemistry was in agreement with histochemically identified type I and type II fibers, respectively.

Ontogeny of the pectoralis muscle in the little brown bat, Myotis lucifugus.

The ontogeny of a primary flight muscle, the pectoralis, in the little brown bat (Myotis lucifugus: Vespertilionidae) was studied using histochemical, immunocytochemical, and electrophoretic techniques. In fetal and early neonatal (postnatal age 1-6 days) Myotis, histochemical techniques for myofibrillar ATPase (mATPase) and antibodies for slow and fast myosins demonstrated the presence of two fiber types, here called types I and IIa. These data correlated with multiple transitional myosin heavy chain isoforms and native myosin isoforms demonstrated with SDS-PAGE and 4% pyrophosphate PAGE. There was a decrease in the distribution and number of type I fibers with increasing postnatal age. At postnatal age 8-9 days, the adult phenotype was observed with regard to muscle fiber type (100% type IIa fibers) and myosin isoform profile (single adult MHC and native myosin isoforms). This "adult" fiber type profile and myosin isoform composition preceded adult function by about 2 weeks. For example, little brown bats were incapable of sustained flight until approximately postnatal day 24, and myofiber size did not achieve adult size until approximately postnatal day 25. Although Myotis pectoralis is unique in being composed of 100% type IIa fibers, transitional fiber types and isoforms were present. These transitional forms had been observed previously in other mammals bearing mixed adult muscle fibers and which undergo transitional stages in muscle ontogeny. However, in Myotis pectoralis, this transition transpires relatively early in development.

Neonatal development of the diaphragm of the horse, Equus caballus.

The diaphragm of neonatal horses is significantly different from the diaphragm of adult horses in terms of histochemical fiber type composition, myosin heavy chain isoform, and native myosin isoform composition. There is a significant increase in the percentage of type I fibers present in the diaphragm with increasing age from birth through about seven months postnatal age. A possible lack of postural tone in the hiatal region of the neonatal diaphragm is suggested to account for increased incidence of vomiting or aspiration pneumonia in younger horses. The isoform data lead to rejection of the hypothesis that the diaphragm of the horse should, as an ungulate, be relatively precocial in its rate of maturation relative to other non-ungulate mammals that have been studied.

Morphological, histochemical, and myosin isoform analysis of the diaphragm of adult horses, Equus caballus.

The horse provides an interesting model for study of the structure and function of the mammalian diaphragm. Multiple regions of diaphragm from seven adult horses were prepared for histochemistry, immunocytochemistry, myosin heavy chain electrophoresis, and native myosin electrophoresis. Two additional adults were dissected to demonstrate myofiber and central tendon morphology and stained for acetylcholinesterase to demonstrate motor endplates. All regions of the adult diaphragm were histochemically characterized by a preponderance of type I fibers with some type IIa fibers. Type IIb fibers were absent in all adult specimens. Myosin heavy chain electrophoresis supported the histochemical study: two isoform bands were present on SDS gels that comigrated at the same rate as rat type I and IIa myosin heavy chain isoforms. No isoform was determined to comigrate with rat type IIb heavy chain isoforms. Native myosin isoform analysis revealed two isoforms that comigrated with rat FM-4 and FM-3 (FM = fast myosin) and two isoforms that comigrated with rat SM-1 and SM-2 (SM = slow myosin) isoforms. In some samples, a third slow native myosin isoform was observed that comigrated at the same rate as the SM-3 of the equine biceps brachii muscle. This doublet (or "triplet") of slow isoforms is unique to some horse muscles compared with other adult animals studied. It is not known if these multiple slow native myosin isoforms confer some functional advantage to the equine muscles. The adult equine diaphragm also differs in its morphology by having a large central tendon compared to that in other mammals, and is predominantly slow in fiber type and myosin isoform composition.

Pectoralis muscle morphology in the little brown bat, Myotis lucifugus: a non-convergence with birds.

Recent studies of muscle architecture demonstrate that many mammalian muscles are composed of short, interdigitating fibers. In addition, the avian pectoralis, a muscle capable of producing high frequency oscillations has been shown to possess a serially arranged pattern of muscle endplate in all sizes of birds studied. The pectoralis muscle of the little brown bat, Myotis lucifugus (Chiroptera: Vespertilionidae), is composed of fairly uniform fibers that span the length of the muscle and is characterized by a zone of motor endplates within the middle third of the muscle. The homogeneous fiber architecture of the bat pectoralis muscle is in contrast to the serial arrangement of endplates (and presumably muscle fibers) in the avian pectoralis in species equivalent in size to Myotis. The short fiber organization and motor endplate pattern observed in most birds is thus not a requisite design for flying vertebrates.

Histochemical and myosin composition of vampire bat (Desmodus rotundus) pectoralis muscle targets a unique locomotory niche.

The vampire bat pectoralis muscle contains at least four fiber types distributed in a nonhomogeneous pattern. One of these fiber types, here termed IIe, can be elucidated only by adenosine triphosphatase (ATPase) histochemistry combined with reactions against antifast and antislow myosin antibodies. The histochemical and immunohistochemical observations indicate a well-developed specialization of function within specific regions of the muscle. In parallel, analyses of native myosin isoforms and myosin heavy chain isoforms indicate two points. First, the histochemical "type IIe" fiber is predominant in cranial portions of the muscle, and myosin extracted from these regions exhibits a unique electrophoretic mobility not observed in the myosin isoforms of more traditional laboratory mammals. Second, the type I fibers are confined to the pectoralis abdominalis muscle and a small adjacent region of the caudal part of the pectoralis. This pattern of type I fiber distribution is considered a derived character state compared to muscle histochemical phenotype and isoform composition in the pectoralis muscles of other phyllostomids we have studied (Artibeus jamaicensis, Artibeus lituratus, Carollia perspicillata). We relate this to the unique locomotory needs of the common vampire bat, Desmodus rotundus.