Histochemical analysis of muscle fiber types of rat superior rectus extraocular muscle

Extraocular muscles (EOMs) represent a distinctive class among mammalian skeletal muscles in exhibiting unique anatomical and physiological properties. To gain insight into the basis for the unique structural/functional diversity of EOM fiber types and to explain their high fatigue resistance, rat superior rectus muscle (SRM) was studied using histochemical techniques. Muscle fibers were typed with regard to their oxidative and glycolytic profiles generated from succinic dehydrogenase (SDH) and phosphorylase activities in combination with their morphologic characteristics. Superior rectus muscle is organized into two layers, a central global layer (GL) of mainly large diameter fibers and an outer C-shaped orbital layer (OL) of principally small diameter fibers. Five muscle fiber types were recognized within the SRM: I, II, III, IV and V. In the global layer, four muscle fiber types were identified: type I (18.25±0.96 μm; 32%) showed intermediate SDH (coarse type) and high phosphorylase activity. Type II fibers (14.45±0.82 μm; 22%) exhibited high SDH (fine type) and intermediate phosphorylase activity. Low SDH (granular type) and high phosphorylase activity were demonstrated by type III fibers (22.65±1.73 μm; 36%). Type IV fibers (26.24±1.32 μm; 10%) were recognized by their low oxidative and glycolytic reactions. In the orbital region, only three muscle fiber types were recognized; fiber types I and II were found to compose approximately two-thirds of the layer. The third orbital fiber type (type V, 10.05±0.99 μm) exhibited low SDH and low phosphorylase profiles. In this paper, the functional significance of the histochemical characteristics of the EOM fiber types is discussed.     

Type-specific changes in fiber morphometry following denervation of canine extraocular muscle.

To investigate the changes that occur in extraocular muscle morphometry following denervation, 12 beagles were subjected to intracranial section of the left oculomotor nerve. The inferior and medial rectus muscles were removed from both orbits of four dogs killed at each 4-week postoperative interval. Fiber diameters and fiber-type ratios of denervated muscles were compared with those from paired muscles in the normal orbit. We found significant, persistent atrophy of the singly innervated fibers in both the global and orbital layers of denervated extraocular muscle. The multiinnervated fibers were predominantly spared from denervation atrophy. We also found a significant increase in the proportion of multiinnervated fibers in the orbital layer only. These results suggest a relative neurotrophic independence of multiinnervated fibers in extraocular muscle.     

Electrical properties and innervation of fibers in the orbital layer of rat extraocular muscles

1. The inferior rectus muscle of rat, one of the extraocular muscles, contains two populations of multiply innervated fibers (MIFs): orbital MIFs, located in the orbital layer of the muscle and global MIFs, found in the global layer. The electrical properties and the responses to nerve stimulation of orbital MIFs were studied with single intracellular electrodes and compared with those of twitch fibers of the orbital layer, MIFs of the global layer, and tonic fibers of the frog. 2. About 90% of the orbital MIFs did not produce overshooting action potentials. In these fibers the characteristics and time course of the responses to nerve stimulation varied along the length of the fibers. Within 2 mm of the end-plate band of the muscle, the responses consisted of several small end-plate potentials (EPPs) and a nonovershooting spike. Distal to 2 mm, the responses in most fibers consisted of large and small EPPs with no spiking response. Some fibers produced very small spikes surmounted on large EPPs. 3. Overshooting action potentials were observed in approximately 10% of the orbital MIFs recorded between the end-plate band and 2 mm distal. The presence or absence of action potentials was not related to the magnitude of the resting potential of the fibers. 4. The threshold of nerve stimulated responses in orbital MIFs was the same as that in orbital twitch fibers. A large number of orbital MIFs had latencies equal to those for the orbital twitch fibers recorded at the same distance from the end-plate band, but the average latency was greater in the MIFs. The latency of orbital MIFs was about one-half of that for the MIFs of the global layer. The values for the effective resistance and membrane time constant of orbital MIFs fell between those for orbital twitch fibers on the one hand, and global MIFs and frog tonic fibers on the other. 5. In order to compare electrical properties with innervation patterns, fibers identified electrophysiologically as orbital MIFs were injected with the fluorescent dye Lucifer yellow and then traced in Epon-embedded, serial transverse sections. In addition to numerous superficial endings distributed along the fibers, a single "en plaque" ending was also found in the end-plate band that resembled the end plates of the adjacent orbital twitch fibers. 6. From these results we conclude that the electrical activity of orbital MIFs varies along the length of the fibers.(ABSTRACT TRUNCATED AT 400 WORDS)     

Histochemistry of mouse extraocular muscle

Summary The organization, histochemical and endplate characteristics, and percentage fiber composition are described for mouse extraocular muscle (EOM). Both recti and obliques, but not the retractor bulbi, have two concentric layers, an inner global and superficial orbital. Three histochemical fiber types, coarse, fine and granular, are present in the EOM. The intermediate diameter coarse fibers are focally innervated and occur in both layers in all muscles. The large diameter granular fibers are focally innervated and occur in all EOM in the global layer. The small diameter fine fibers have multierminal endplates and occur in both layers of the recti. Fine fibers are not seen in the obliques or retractor bulbi. Focal endplates are confined to a broad diagonal band across the middle one third of the muscle, whereas multiterminal endplates are scattered throughout the length of the muscle.This research was carried out during the tenure of a Postdoctoral Fellowship from the Muscular Dystrophy Association     

Systematic variation in myosin expression along extraocular muscle fibres of the adult rat

Summary Monoclonal antibodies (McAB) specific for fast (C14) and slow (S58) myosin, and a myosin antigenically similar to neonatal/embryonic myosin in mammals (ALD180), were used to characterize the myosin distribution in orbital layer fibres of rat extraocular muscles (EOM) in relation to innervation patterns. The orbital layer is composed of both singly-innervated (SIF) and multiply-innervated (MIF) fibres. The SIFs have the characteristics of twitch fibres, while the MIFs, in addition to possessing many small endings characteristic of tonic fibres, also have an en-plaque-like innervation in the endplate band resembling that of the adjacent SIFs. Myosin expression in MIFs and SIFs is unusual and varies systematically along the length of the fibres. Both SIFs and MIFs label with ALD180, but this labelling is absent in both fibre types in the endplate band region, where all fibres label with C14. Distally and also proximally to the endplate band, SIFs label with both ALD180 and C14, while the MIFs, innervated by many small, superficial endings in these regions, label with ALD180 only. This pattern of myosin expression could also be demonstrated in isolated fibres. The results are discussed in relation to the hypothesis that both populations of orbital layer fibres express constitutively both fast and the neonatal-like myosin, and that superimposed on this constitutive expression twitch or tonic innervation acts locally to selectively suppress either neonatal-like or fast myosin, respectively.     

Internal structure of cat extraocular muscle.

Teasing preparations of cat extraocular muscles (EOM) were used to study the arrangement of muscle fibers and the distribution of the different cholinesterase-positive sites, i.e. (1) large motor endplates, (2) small motor endings of the 'en grappe' type, (3) myotendinous junctions and (4) myomyous junctions. The distribution of these cholinesterase-positive structures gives clear evidence of a complex muscle architecture of cat EOM. In the global layer of cat EOM, only multiply innervated muscle fibers run the whole length of the muscle. The focally innervated muscle fibers are generally shorter; they are usually arranged in series of two to three fibers being interconnected by myomyous junctions. Moreover, muscle fiber splitting is frequently present resulting in a netlike arrangement of muscle fibers. Most of the myomyous junctions occur between focally innervated muscle fibers, but also end-to-side connections of focally to multiply innervated muscle fibers are observed; multiply innervated muscle fi0ers have not been found connected to each other. In this layer, large motor endplates are distributed in several bands between origin and insertion. In the orbital layer all muscle fibers run from tendon to tendon, focally as well as multiply innervated ones. Here, large motor endplates are confined to a band in the middle of the muscle, and myomyous junctions are generally absent. Some functional implications of this complex architecture of cat EOM are discussed.     

The postnatal development of the inferior oblique muscle of the cat

Transversal sections of the inferior oblique muscle from kittens of various ages and from adult cats were stained for myofibrillar ATP-ase at Ph 9.4 and 4.35, succinic dehydrogenase (SDH) and fat. The same muscles had previously been submitted to studies of contractional and fatigue properties. With ATPase the fibers could be differentiated into types I, II and II C. The percentages of each fiber type remained approximately the same from birth onwards. In the inner, global layer of the muscle, type I fibers showed a linear increase in size, but types II and IIC fibers an accelerated growth after age 20 weeks. In the outer, orbital layer, where no type I fibers were seen, types II and IIC fibers showed the same growth pattern as in the global layer. The fiber content of SHD and fat was low at birth but increased after two weeks of age. Type I fibers were poor and type IIC fibers rich in SDH and fat. Type II fibers showed varying amounts of these substances. Provided that type 1 fibers are slow, type II fast and type II C intermediate in speed of contraction, like in other muscles, the findings on fiber growth and SDH content seem to support the idea that slow, fatigue resistant components in eye muscles reach maturity earlier than fast components.     

大鼠眼上直肌神经的生后发育

取生后两天、两周和两月的雄性Ｗｉｓｔａｒ大鼠双眼上直肌，进行硝酸银染色和乙酰胆碱酯酶染色，用光镜观察大鼠眼上直肌神经在生后的发育情况。在大鼠生后两天时，动眼神经的分支从眼上直肌的中后三分之一交界处由眶面进入该肌，它向该肌的球面区发出细小的神经纤维束，这些神经束抵达同一条肌纤维上。眶面层神经纤维细小，单条并且平行于肌纤维方向走行。此时，眼上直肌对乙酰胆碱酯酶染色着色浅，反应区未形成某一特定轮廓，说明此时运动神经的发育是不成熟的。在生后两周时，动眼神经的分支开始向眼上直肌的眶面层发出神经纤维；球面区中的神经纤维有明显的粗细两种。这时眼上直肌中的运动神经轴索对乙酰胆碱酯酶染色反应，出现葡萄状运动神经末梢及斑点状运动终板的雏形。生后两月时，眼上直肌中出现了肌梭及典型的葡萄状运动神经末梢和斑点状运动终板。生后两周是大鼠睁眼初期，由此可见，大鼠从闭眼至睁眼的时期，是运动神经发育的关键时期，此时，运动神经形成一些特定神经末梢，而睁眼后运动神经的发育主要在此基础上运动神经末梢日趋完善和成熟。     

Classification and Development of Myofiber Types in the Superior Oblique Extraocular Muscle of Chicken

Precise control of contractile force of extraocular muscles is required for appropriate movements and alignment of the eyes. It is unclear how such precise regulation of contractile force is achieved during development and maturation. By using the posthatch chicken as a model, we describe and quantify critical parameters of the developing superior oblique extraocular muscle from hatching to 16 weeks of age, including contractile force, muscle mass, myofiber diameters, classification of fiber types, and distribution and quantification of mitochondria. Analysis at the light‐ and electron microscopic levels shows that chicken myofiber types largely correspond to their mammalian counterparts, with four fiber types in the orbital and four types in the global layer. Twitch tension muscle force and muscle mass gradually increase and stabilize at approximately 11 weeks. Tetanic tension continues to increase between 11 and 16 weeks. Myofiber diameters in both the orbital and global layer increase from hatching to six weeks, and then stabilize, whereas the myofiber number is constant after hatching. This finding suggests that muscle mass increases during late maturation due to increasing fiber length rather than fiber diameter. Quantitative ultrastructural analysis reveals continuing changes in the composition of the four muscle fiber types, suggesting ongoing fiber type conversion or differential replacement of myofiber types. Muscle fiber composition continues to change into late juvenile and adult age. Our study provides evidence for gradual, incremental, and continuing changes in avian myofiber composition and function that is similar to postnatal oculomotor maturation in visually oriented mammals such as kitten. Anat Rec, 2007. © 2007 Wiley‐Liss, Inc.     

Mitochondrial morphometrics of histochemically identified human extraocular muscle fibers

Three fiber types--coarse, granular, and fine--were readily identified in histochemical cryostat sections of human extraocular muscle (EOM). The cryostat retrieval method was utilized to identify these three fiber types in serial electron microscopic thin sections. Using morphometric techniques, five mitochondrial variables (mitochondrial volume fraction, mitochondrial profile size, mitochondrial profile density, and clusters of two or of three or more mitochondrial profiles) were determined for a total of 162 histochemically identified fibers from two regions (orbital and global zones) from six EOMs. Coarse fibers had numerous large-sized mitochondrial profiles, often occurring in clusters. Granular fibers had fewer and smaller-sized profiles scattered across the fiber. Fine fibers had the most numerous, but smallest-sized mitochondrial profiles. Despite significant differences in group (fiber types) means for the mitochondrial variables, no single variable was sufficient for separating fiber types into distinct populations. Although a scattergram plot of two variables was sufficient to separate orbital zone fibers, a computer-generated, multivariate discriminant analysis was needed to separate the global zone fibers into distinct populations. These results will aid future studies on normal and pathological human EOM by providing a morphometric basis for identifying fiber types in the orbital and global zones.     

Spatial and temporal patterns of myosin heavy chain expression in developing rat extraocular muscle

Summary The present study describes transitions in myosin heavy chain expression in the extraocular muscles of rats between the ages of E17 and adult. The unique phenotype of the extraocular muscle is reflected in its fibre type composition, which is comprised by six distinct profiles, each defined by location (orbital versus global layer) and innervation pattern (single versus multiple terminals). During extraocular muscle myogenesis, developmental myosin heavy chains were expressed in both primary and secondary fibres from embryonic day E17 through the first postnatal week. At this time, the downregulation of developmental myosin heavy chain isoforms began in the global layer in a fibre type-specific manner, reaching completion only after the first postnatal month. By contrast, developmental isoforms were retained in the overwhelming majority of orbital layer fibres into adulthood and expressed differentially along the length of these fibres. Fast myosin heavy chain was detected pre- and postnatally in developing secondary fibres and in all of the singly innervated fibre types and one of the multiply innervated fibre types in the adult. As many as four fast isoforms were detected in maturing extraocular muscle, including the extraocular muscle-specific myosin heavy chain. Slow myosin heavy chain was expressed in primary fibres throughout development and in one of the multiply innervated fibre types in the adult. In contrast, the pure fast-twitch retractor bulbi initially expressed slow myosin heavy chain in fibres destined to switch to the fast myosin heavy chain developmental programme. Based upon spatial and temporal patterns of myosin heavy chain isoform transitions, we suggest that epigenetic influences, rather than purely myogenic stage-specific factors, are critical in determining the unique extraocular muscle phenotype.     

Localization of motoneurons innervating the extraocular muscles in the chameleon (Chamaeleo chameleon)

The topography and localization of motoneurons innervating the six extraocular muscles in the chameleon (Chamaeleo chameleon) was studied following HRP injection in each of these individual muscles. Four muscles were innervated ipsilaterally: medial rectus, inferior rectus, inferior oblique and lateral rectus. The medial rectus muscle was innervated by the dorsomedial part of the oculomotor nucleus. The innervation to the inferior rectus muscle arose from the lateral part of the intermediate oculomotor subnucleus, which extended to the lateral part of the dorsal subdivision. The lateral rectus muscle was innervated by the abducens nucleus, which was composed by two subgroups of labeled cells, respectively observed in the principal and accessory abducens subnuclei, whereas efferents to the inferior oblique muscle originated from both the ventral and intermediate oculomotor subnuclei. The contralateral pattern consisted of motoneurons innervating the superior rectus and the superior oblique that were located respectively in the caudal portion of the ventral oculomotor nucleus and in the trochlear nucleus. These results confirmed data reported in most vertebrate species, and were discussed from a comparative and functional point of view.     

Motoneuronal innervation and mechanical properties of extraocular muscles in the catfish, (Ictalurus punctatus)

Mechanical characteristics and electrical activity were studied in the extraocular muscles of the catfish, Ictalurus punctatus. The contractile properties were determined by stimulation of the individual muscle nerve branches to lateral and medial rectii and the superior and inferior oblique muscles. The speed of contraction was higher than in most other fish muscle, with a twitch contraction time of about 12 ms and a tetanus fusion frequency of 150–170 Hz in all four eye muscles. The fatigue resistance was also high. These properties were the same in fully innervated and partially innervated muscle, largely irrespective of what part of the muscle that was activated. Although different fibre types are known to exist in fish extraocular muscle, it was not possible to obtain functional separation of the mechanical force profile even in the lateral rectus with two distinct motoneuronal innervations. We suggest that polyneuronal innervation of the muscle fibres produces the mechanical responses. Since EMG activity during spontaneous eye movements was similar in the global and the orbital parts of the muscle, all types of fibres in fish extraocular muscle are probably recruited for all types of eye movements.     

Complex three-dimensional patterns of myosin isoform expression: differences between and within specific extraocular muscles

Because complex structural differences in adult extraocular muscles may have physiological and pathophysiological significance, the three-dimensional pattern of myosin heavy chain (MHC) isoform expression within the orbital and global layers of the muscle bellies compared with the distal tendon ends was quantitatively assessed. Three of the six extraocular muscles of adult rabbits were examined for immunohistologic expression of all fast, fast IIA/X, slow, neonatal and developmental MHC isoforms. The percentages of myofibers positive for each of these 5 myosin isoforms were determined in the orbital and global layers. There were relatively similar patterns of fast and slow MHC expression in the orbital and global layers of each of the three muscles examined. There were high levels of developmental MHC in the orbital layers, but significantly fewer developmental MHC positive myofibers in the global layer. The most variable expression was found with the neonatal MHC. There were significant differences between the longitudinal expression of the various isoforms in the middle of each muscle compared with the tendon end. In the orbital layer of all three muscles examined, the large numbers of fibers positive for fast MHC in the middle of the muscle dramatically decreased at the tendon end, with a concomitant increase in expression of slow myosin. There was a greater number of developmental MHC-positive myofibers at the tendon end than in the middle of the muscle in all three muscles examined. In the global layer, the IIA/X-positive myofibers comprised only half of the total number of fast-positive myofibers whereas in the orbital layer they comprised all or almost all of the fast positive myofibers. The configuration of the extraocular muscles is more complex than might be indicated by previous studies. The lateral rectus muscle had the most individual pattern of MHC expression when compared with the inferior rectus and inferior oblique muscles. Together with dramatic cross-sectional MHC fiber type differences between the orbital and global layers of the muscles, there are pronounced longitudinal differences in the proportions of myofibers expressing these five MHC isoforms in the middle region of the muscles and those in the distal tendon ends. This longitudinal progression appears to occur both within single myofibers, as well as within the series of myofibers that comprise the length of the muscle. We also confirm that the number of myofibers is reduced at the tendonous end while the cross-sectional area of each of the remaining myofibers is proportionally increased with regard to those in the muscle belly. Future studies may yet require two additional schemes for anatomic classification of the named extraocular muscles. One will be based on immunohistochemical features of their constituent myofibers as a supplement to classifications based on their electron microscopic appearance, innervation patterns or relative position with regard to the globe and orbit. Another will be based on the proportional length and longitudinal position of individual myofibers within an individual extraocular muscle.     

Localization of motoneurons innervating the extraocular muscles in the chameleon (Chamaeleo chameleon)

The topography and localization of motoneurons innervating the six extraocular muscles in the chameleon (Chamaeleo chameleon) was studied following HRP injection in each of these individual muscles. Four muscles were innervated ipsilaterally: medial rectus, inferior rectus, inferior oblique and lateral rectus. The medial rectus muscle was innervated by the dorsomedial part of the oculomotor nucleus. The innervation to the inferior rectus muscle arose from the lateral part of the intermediate oculomotor subnucleus, which extended to the lateral part of the dorsal subdivision. The lateral rectus muscle was innervated by the abducens nucleus, which was composed by two subgroups of labeled cells, respectively observed in the principal and accessory abducens subnuclei, whereas efferents to the inferior oblique muscle originated from both the ventral and intermediate oculomotor subnuclei. The contralateral pattern consisted of motoneurons innervating the superior rectus and the superior oblique that were located respectively in the caudal portion of the ventral oculomotor nucleus and in the trochlear nucleus. These results confirmed data reported in most vertebrate species, and were discussed from a comparative and functional point of view. Accepted: 18 June 1999     

Identification of motoneurons supplying multiply- or singly-innervated extraocular muscle fibers in the rat

In mammals, the extraocular muscle fibers can be categorized in singly-innervated and multiply-innervated muscle fibers. In the monkey oculomotor, trochlear and abducens nucleus the motoneurons of multiply-innervated muscle fibers lie separated from those innervating singly-innervated muscle fibers and show different histochemical properties. In order to discover, if this organization is a general feature of the oculomotor system, we investigated the location of singly-innervated muscle fiber and multiply-innervated muscle fiber motoneurons in the rat using combined tract-tracing and immunohistochemical techniques. The singly-innervated muscle fiber and multiply-innervated muscle fiber motoneurons of the medial and lateral rectus muscle were identified by retrograde tracer injections into the muscle belly or the distal myotendinous junction. The belly injections labeled the medial rectus muscle subgroup of the oculomotor nucleus or the greatest part of abducens nucleus, including some cells outside the medial border of abducens nucleus. In contrast, the distal injections labeled only a subset of the medial rectus muscle motoneurons and exclusively cells outside the medial border of abducens nucleus. The tracer detection was combined with immunolabeling using antibodies for perineuronal nets (chondroitin sulfate proteoglycan) and non-phosphorylated neurofilaments. In monkeys both antibodies permit a distinction between singly-innervated muscle fiber and multiply-innervated muscle fiber motoneurons. The experiments revealed that neurons labeled from a distal injection lack both markers and are assumed to represent multiply-innervated muscle fiber motoneurons, whereas those labeled from a belly injection are chondroitin sulfate proteoglycan- and non-phosphorylated neurofilament-immunopositive and assumed to represent singly-innervated muscle fiber motoneurons. The overall identification of multiply-innervated muscle fiber and singly-innervated muscle fiber motoneurons within the rat oculomotor nucleus, trochlear nucleus, and abducens nucleus revealed that the smaller multiply-innervated muscle fiber motoneurons tend to lie separate from the larger diameter singly-innervated muscle fiber motoneurons. Our data provide evidence that rat extraocular muscles are innervated by two sets of motoneurons that differ in their molecular, morphological, and anatomical properties.     

Ultrastructural and metabolic profiles on single muscle fibers of different types after hindlimb suspension in rats

The relationships between ultrastructural and metabolic profiles in different types of single muscle fiber after hindlimb suspension in rats were examined. Glycolytic (lactate dehydrogenase, LDH; phosphofructokinase, PFK) and oxidative (succinate dehydrogenase, SDH; malate dehydrogenase, MDH) enzyme activities in extensor digitorum longus (EDL) and soleus (SOL) muscles were measured. Relative mitochondrial and lipid droplet volumes were also measured in single muscle fiber of different types. Glycolytic enzyme activity in EDL muscle and oxidative enzyme activity in soleus muscle decreased following suspension for 2 weeks. LDH and PFK activities in fast-twitch (FG, fast-twitch glycolytic; FOG, fast-twitch oxidative glycolytic) fibers and oxidative enzymes in FOG and FG fibers decreased following suspension. Relative mitochondrial volume decreased significantly in all types (SO, slow-twitch oxidative; FOG, and FG) of fibers following suspension. The mitochondrial volume in SO fiber of the control group was significantly (p less than 0.01) higher than that of suspended group; however, SDH and MDH activities were not different between the control and suspended groups. The structural and metabolic changes following hindlimb suspension were influenced by different factors, respectively. Changes in ultrastructural and metabolic profiles in response to the hindlimb suspension differed according to the type of fibers.     

Ultrastructural observations on muscle spindles in extraocular muscles of pig

Human and some mammals such as the sheep, goat and domestic and wild pigs have more or less muscle spindles in the extrinsic eye muscles, especially the domestic pigs having abundant muscle spindles (Matsuyama, 1987). The muscle spindles play a large role in maintaining the stable visual posture of the eyeballs. To define the morphological properties of the muscle spindles relative to the eye movement, the ultrastructure of the spindles was investigated in 6 extraocular muscles of the pigs by electron microscopy. The muscle spindles in the pig extraocular muscles consist of 4 to 5 intrafusal muscle fibers, one of which is nuclear bag fiber and 3 to 4 are nuclear chain fibers. The outer capsule is thin, composing of few layers, and the inner capsule ramifying to enwrap the individual fiber, accompanied by the medullated and unmedullated nerve fibers and blood capillaries. The nuclear bag fiber, 14 micron in diameter, is innervated by the atypical annulospiral sensory terminals and the chain fiber by the typical annulospiral terminal packed with mitochondria and microvesicles. The intrafusal fibers are innervated by the flower-spray sensory terminals anchoring deeply into the sarcoplasma, having abundant neurotubules and few mitochondria. The gamma motor end-plates have a relatively smooth synaptic cleft with a width of 70 nm and synaptic boutons containing few synaptic vesicles, sometimes, revealing a shallow fold of postsynaptic sarcolemma and abundant synaptic vesicles. The alpha motor end-plates reveal a relatively smooth synaptic cleft with a width of 80 nm, sometimes with a rough postsynaptic infolding, and boutons containing few synaptic vesicles and small-sized mitochondria. The satellite cells are innervated by the sensory terminals in various ways. The muscle spindles in the pig extraocular muscles are found to be much simpler in structure than those in the other antigravity muscles of the body. Their ultrastructure seems to reflect the morphological adaptation relative to the eyebal movement.     

Architecture and fiber type of the pyramidalis muscle

The paired pyramidalis muscles are small triangular-shaped muscles that lie between the anterior surface of the rectus abdominus and the posterior surface of the rectus sheath. The precise function of pyramidalis muscles is unclear, but together the muscles are thought to tense the linea alba. The muscles are not always present, or are often unilateral, and vary greatly in size. Their wider inferior margins attach to the pubic symphyses and pubic crests, whereas their narrow superior margins attach to the linea alba. The gross anatomy and innervation of the pyramidalis muscles has been described by others, but their architecture and fiber type have not been determined in previous publications. The purpose of the present paper was therefore to investigate these parameters and place the findings into context for the literature available on this muscle. An example of bilateral pyramidalis muscles was recently encountered in a male cadaver that provided ample tissue for an analysis of its architecture and fiber type. The muscle mass, muscle length, fiber length, and pennation angle of muscle fibers were measured to ascertain physiological cross-sectional area and thereby estimate force production. Fiber type composition was also examined using immunofluorescent labeling. The results show that this is a muscle of mixed fiber type composition, similar to the rectus abdominus, and that the estimated forces generated by this muscle are relatively small.     

Morphological variability and specializations in bovine extraocular muscle spindles

Bovine extraocular muscles were examined to determine whether the structure of their muscle spindles was notably different from those commonly encountered in mammalian limb muscles. Extraocular muscle spindles on the whole were shorter, and intrafusal fiber counts/spindle were more variable than in somatic muscles. No pronounced nuclear bags were seen in intrafusal fibers. Based on cross-sectional areas, intrafusal fibers in extraocular muscles could be loosely categorized as small or large types. Small fibers expressed more neonatal/fast myosin heavy chain and less embryonic myosin heavy chain than large fibers. When incubated for myosin ATPase, about 70% of the large fibers and 15% of the small fibers in spindles presented profiles that were characteristic of type I extrafusal fibers, and not of nuclear bag or nuclear chain fibers. The ratio of number of small intrafusal fibers to number of large intrafusal fibers in extraocular spindles was on average greater than the ratio of nuclear chain fibers to nuclear bag fibers that is typical for limb spindles of rodents and cats. Structural modifications at muscle spindle sensory regions, extrafusal-like fibers and intrafusal-like fibers with few equatorial nuclei and many myofibrils, may produce distinct afferent signals that are appropriate for sensorimotor integration in the specialized extraocular muscles.