Origin of intrafusal fibers from a subset of primary myotubes in the rat

S46, a monoclonal antibody (mAb) specific for the SM-1 and SM-2 isoforms of avian slow myosin heavy chains (MHC), was used to study the earliest stages of development of intrafusal fibers in muscle spindles of the rat hindlimb. Spindles formed only in the regions of fetal muscles that contained primary myotubes reactive to mAb S46, such as the axial region of the tibialis anterior muscle. The first intrafusal fiber to form, the nuclear bag₂ fiber, originated from within the population of S46-reactive primary myotubes. Binding of mAb S46 by myotubes giving rise to the bag₂ fibers preceded the appearance of encapsulated spindles in the muscles by electron microscopy. However, reactivity to S46 intensified in the myotubes transforming into bag₂ fibers after the innervation of the fibers by afferents, and dissipated in myotubes differentiating into slow-twitch (type I) extrafusal fibers. Thus, afferents may enhance intrafusal expression of the MHC isoform reactive to mAb S46. The pattern of S46 binding to nuclear bag and chain intrafusal fibers in both developing and adult spindles was the same as that reported for the mAb ALD19, suggesting that both antibodies bind to the same MHC isoform. This isoform is probably a developmental form of slow myosin, because it was transiently expressed during the development of type I extrafusal fibers. The origin of bag₂ intrafusal and type I extrafusal fibers from a bipotential subpopulation of primary myotubes reactive to mAb S46 correlates with the location of muscle spindles in the slow regions of muscles in adult rat hindlimbs.     

Quantitative and histochemical aspects of the differentiation of muscle spindles in the anterior latissimus dorsi of the developing chick

Summary The differentiation of muscle spindles has been investigated both quantitatively and histochemically in the slow anterior latissimus dorsi (ALD) muscle during embryonic and post-hatching development of the chick. The first spindles are detected by the 13th day in ovo, and a rapid increase in number takes place until the 15th of embryonic life. Two histochemical fibre types of intrafusal fibres are distinguished as early as the 13th day of embryonic development when myofibrillar ATPase activity is demonstrated after acid preincubation. In ALD muscle from post-hatched animals, two intrafusal fibre types are also distinguished by histochemistry: one fibre type is characterized by an acid and alkali-stable myofibrillar ATPase activity while this activity is acid-labile and alkali-stable in the other type. As far as the properties of myofibrillar ATPase are concerned, such types of intrafusal fibres resemble the extrafusal and fibre types defined recently in ALD muscle.The development of spindles in the slow-tonic ALD muscle is compared to the differentiation of spindles in the fast-twitch posterior latissimus dorsi (PLD) muscle of the chick which has been previously described (Toutant et al. 1981).     

Sequences of intrafusal fiber formation are muscle-dependent in rat hindlimbs

A rat muscle spindle typically contains four intrafusal fibers — one nuclear bag₂, one nuclear bag₁ and two nuclear chain fibers. We compared the sequence of formation of the three intrafusal fiber types among the tibialis anterior (TA), soleus (SOL) and medial gastrocnemius (MG) muscles using immunocytochemistry of spindle-specific myosin heavy chain isoforms. Spindles of the TA began to differentiate earlier and acquired the full complement of intrafusal fibers sooner than spindles of the SOL or MG muscles. At the onset of spindle assembly, the intrafusal myotubes expressed myosin heavy chains similar to those expressed by extrafusal myotubes. The first intrafusal myotube then differentiated into the bag₂ fiber regardless of the muscle. However, the fate of the second-forming intrafusal myotube varied among the muscles studied. It usually differentiated into a chain fiber in the TA, into a bag₁ fiber in the SOL, and into either a bag₁ or a chain in the MG. The fate of the third-forming intrafusal myotube was reciprocal to that of the second; i.e. in those spindles in which the bag₁ fiber was second to form, a chain was third, and vice versa. The fourth and last intrafusal myotube gave rise to a chain fiber. The inter- and intramuscular variability in the fate of intrafusal myotubes of the second and third generation argues against the existence of a program intrinsic to the myotubes that would mandate their differentiation along specific paths. Rather, an extrinsic regulatory factor, probably associated with the primary afferent neuron, may govern differentiation of pluripotential myotubes into particular types of intrafusal fiber. The fate of the intrafusal myotubes might then depend on the timing of the regulatory effect of afferents relative to the stage of development of the intrafusal bundle.     

Postnatal expression of myosin heavy chains in muscle spindles of the rat

Summary The immunocytochemical expression of two myosin isoforms in intrafusal muscle fibers was examined in soleus muscles of neonatal (zero to six days postpartum) and adult rats. Monoclonal antibodies specific for myosin heavy chains of the slow-tonic anterior latissimus dorsi (ALD58) and fast-twitch pectoralis (MF30) muscles of the chicken were used. In adults ALD58 bound to the intracapsular regions of bag₁ and bag₂ fibers and MF30 bound to the intracapsular regions of bag₂ and chain fibers. The extracapsular regions of intrafusal fibers and all extrafusal fibers did not react to ALD58 or MF30. Bag₁ and bag₂ fibers of neonatal rats expressed immature myosin patterns but chain fibers did not. The adult pattern of immunoreactivity of intrafusal fibers developed by the fourth postnatal day, when the patterns of motor but not sensory innervation in the spindle are still immature. Data suggest that the expression and maintenance of the specific anti-myosin immunoreactivity of intrafusal fibers during postnatal development of rat spindles is dependent upon sensory but not motor innervation. Moreover, afferents might regulate the gene expression responsible for synthesis of myosins isoforms specific to intrafusal fibers only in those myonuclei located within the capsule, but not in the myonuclei in extracapsular regions of intrafusal fibers.     

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.     

Differential effects of neonatal denervation on intrafusal muscle fibers in the rat

The response of developing muscle spindles to denervation was studied by sectioning the nerve to the medial gastrocnemius muscle of rats at birth. The denervated spindles were examined daily throughout the first postnatal week for changes in ultrastructure and expression of several isoforms of myosin heavy chain (MHC). Each of the three different types of intrafusal muscle fiber exhibited a different response to denervation. Within 5 days after the nerve section nuclear bag₂ fibers degenerated completely; nuclear bag₁ fibers persisted, but ceased to express the spindle-specific slow-tonic MHC isoform and thereby could not be differentiated from extrafusal fibers; nuclear chain fibers did not form. The capsules of spindles disassembled, hence spindles or their remnants could no longer be identified 1 week after denervation. Neonatal deefferentation has little effect on these features of developing spindles, so removal of afferent innervation is presumably the factor that induces the loss of spindles in denervated muscles. Degeneration of the bag₂ fiber, but not bag₁ or extrafusal fibers, reflects a greater dependence of the bag₂ fiber than the bag₁ fiber on afferent innervation for maintenance of its structural integrity. This difference in response of the two types of immature bag fiber to denervation might reflect an origin of the bag₂ fibers from a lineage of myogenic cells distinct from that giving rise to bag₁ or extrafusal fibers, or a difference in the length of contact with afferents between the two types of bag fiber prior to nerve section.     

Structure, distribution and innervation of muscle spindles in avian fast and slow skeletal muscle

Muscle spindles in 2 synergistic avian skeletal muscles, the anterior (ALD) and posterior (PLD) latissimus dorsi, were studied by light and electron microscopy to determine whether morphological or quantitative differences existed between these sensory receptors. Differences were found in the density, distribution and location of muscle spindles in the 2 muscles. They also differed with respect to the morphology of their capsules and intracapsular components. The slow ALD possessed muscle spindles which were evenly distributed throughout the muscle, whereas in the fast PLD they were mainly concentrated around the single nerve entry point into the muscle. The muscle spindle index (number of spindles per gram wet muscle weight) in the ALD was more than double that of its fast-twitch PLD counterpart (130.5±2.0 vs 55.4±2.0 respectively, n=6). The number of intrafusal fibres per spindle ranged from 1 to 8 in the ALD and 2 to 9 in the PLD, and their diameters varied from 5.0 to 16.0 μm and 4.5 to 18.5 μm, respectively. Large diameter intrafusal fibres were more frequently encountered in spindles of the PLD. Unique to the ALD was the presence of monofibre muscle spindles (12.7% of total spindles observed in ALD) which contained a solitary intrafusal fibre. In muscle spindles of both the ALD and PLD, sensory nerve endings terminated in a spiral fashion on the intrafusal fibres in their equatorial regions. Motor innervation was restricted to either juxtaequatorial or polar regions of the intrafusal fibres. Outer capsule components were extensive in polar and juxtaequatorial regions of ALD spindles, whereas inner capsule cells of PLD spindles were more numerous in juxtaequatorial and equatorial regions. Overall, muscle spindles of the PLD exhibited greater complexity with respect to the number of intrafusal fibres per spindle, range of intrafusal fibre diameters and development of their inner capsules. It is postulated that the differences in muscle spindle density and structure observed in this study reflect the function of the muscles in which they reside.     

Morphological and histochemical differentiation of intrafusal fibres in the posterior latissimus dorsi muscle of the developing chick

Summary Morphological and histochemical differentiation of neuromuscular spindles was studied in the posterior latissimus dorsi (PLD) of the chick during embryonic and post-hatching development. A rapid increase in the number of spindles takes place between the 13th and 15th of embryonic life. By the 15th day in ovo, the spindle capsule appears filled with numerous contiguous cells. Large sensory endings and small primitive motor endings are observed on intrafusal fibres. Ultrastructural observations of the nerve supply of the spindles confirm that each developing spindle receives one thick Ia axon with one to three thin axons. The intracapsular space differentiates by the 17th day of embryonic development. All intrafusal fibres are morphologically of the nuclear-chain type, while two fibre types are distinguished as early as the 14th day of embryonic life, when myofibrillar ATPase activity is demonstrated after acid preincubation. These two histochemical types of intrafusal fibres are also described in the adult. The relation between these two histochemical types and different functional activity of intrafusal fibres is suggested.     

Effects of 14 days of microgravity on fast hindlimb and diaphragm muscles of the rat

The purpose of the present study was to determine the effects of 14 days of microgravity on specific rat fast-twitch muscles, and to compare these data with previous data from rat fast-twitch muscles exposed to microgravity for 10 days (Kraemer et al. 2000). Hindlimb muscles containing predominately fast fibers [extensor digitorum longus (EDL), superficial “white” (GSW) and deep “red” (GDR) gastrocnemius] and the diaphragm (DIA) were removed from flight and ground-based control animals and analyzed for: muscle mass, fiber type distribution, cross-sectional area, and myosin heavy chain (MHC) isoform content. Gravitational unloading for 14 days caused significant decreases in muscle mass (8–9%) and cross-sectional area of almost all fiber types (10–35%) from both EDL and gastrocnemius muscles. However, microgravity had little effect on fiber type composition in these muscles with significant changes occurring only in the EDL type IID fiber population (9.5% decrease). Similarly, relative MHC isoform content was only slightly altered by exposure to microgravity (increased content of MHCIIa in flight EDL). No changes in area, fiber type percentages, or MHC isoform content were detected in the DIA following the 14-day spaceflight. Similar to data gathered following a 10-day spaceflight (Kraemer et al. 2000), the 14-day flight did not appear to cause significant slow-to-fast (I → IIA) or fast-to-faster (IIA → IID → IIB) transformations in hindlimb muscles containing predominantly fast-twitch fibers. However, the longer period of gravitational unloading did result in additional loss in muscle fiber cross-sectional area with involvement of more major fiber types.     

Axon contacts and acetylcholinesterase activity on chicken intrafusal muscle fiber types identified by their myosin heavy chain composition

Summary Muscle spindles of 8-week old chicken tibialis anterior muscles were examined to determine if specific intrafusal fiber types were also characterized by differences in motor innervation. Incubation with a monoclonal antibody against myosin heavy chains permitted the identification of strongly reactive, moderately reactive and unreactive intrafusal fibers. The innervation of each fiber type was evaluated in silver-impregnated sections, and in sections incubated with a monoclonal antibody against acetylcholinesterase. There was no acetylcholinesterase activity at the midequator of any fiber. At the juxtaequator and at the pole strongly reactive fibers typically exhibited fewer axon contacts and less acetylcholinesterase activity than unreactive and moderately reactive fibers. Differences were also recognized at neuromuscular junctions in the size and shape of acetylcholinesterase-positive sites. At the juxtaequator and at the pole strongly reactive fibers and moderately reactive fibers displayed significantly more small, dot-like acetylcholinesterase sites than unreactive fibers. On the contrary, the greatest number of larger, stout sites was found on unreactive fibers and the least number on strongly reactive fibers. Moderately reactive fibers took an intermediate position. The results indicate that myosin heavy chain-based chicken intrafusal fiber types are also set apart by differences in innervation.     

Correlation between percentage fiber type area and myosin heavy chain content in human skeletal muscle

Histochemical methods are routinely used to delineate skeletal muscle fiber types. In the present investigation, this qualitative determination of fiber type composition was compared to the electrophoretically determined myosin heavy chain (MHC) content from a large number of human muscle biopsy samples. Biopsies were taken from the vastus lateralis muscle at the beginning and every 2 weeks during 8 weeks of highi-ntensity resistance training from men (n = 13) and woman (n = 8). Muscle was also extracted from nontraining men (n = 7) and women (n = 5) at the same periods. Six muscle fiber types (I, IC, IIAC, IIA, IIAB, and IIB) were determined using basic myofibrillar adenosine triphosphatase histochemistry. Cross-sectional areas were determined for the three major fiber types (I, IIA, and IIB) and used to calculate the percentage area of these types. Electrophoretic techniques were used to separate and quantify the percentage MHC content in these same biopsy samples, and these data were then used to compare with the percentage fiber type area. Correlation analyses suggest a relationship between the histochemically assessed percentage fiber type area and the electrophoretically assessed MHC content in human limb musculature. However, because of possible histochemical misclassification of some fibers (especially in trained muscle) both techniques may be essential in yielding important information about fiber type composition and possible fiber type transformations.     

Myelinated nerve fiber supply and muscle spindles in the respiratory muscles of cat: Quantitative study

Summary The present study was undertaken to provide quantitative data on the myelinated fibers of the phrenic and intercostal nerves and the number of spindles in the main respiratory muscles of the cat.The myelinated component of the phrenic and intercostal nerves was studied in the cat. Histograms of sequency distributions as a function of nerve fiber diameter were established for normal nerves. Certain nerves were then examined 35 to 40 days after excision of the dorsal spinal ganglia. The muscle spindles of the corresponding muscles were counted and localized, and, on the basis of several morphological criteria, were classified with those usually described in the interosseous muscles.The study of the nerves, as that of the spindles, demonstrates clear differences of proprioceptive innervation among the respiratory muscles. The lateral part of the diaphragm and the Triangularis sterni have practically no spindles. The external muscles of the first thoracic spaces are very rich in spindles. Respiratory muscles can be ranged in an almost continuous manner between these two extremes.     

Motor and sensory innervation of muscle spindles in the neonatal rat

Summary Neural and muscular elements of three muscle spindles from the soleus muscles of 4-day-old rats were reconstructed by electron microscopy of skip-serial transverse ultrathin sections. Each spindle contained four encapsulated intrafusal fibers, including a minimum of one bag₁, one bag₂ and one chain fiber. The fibers were innervated by unmyelinated motor and sensory axons. The primary and secondary afferents approached the spindles as single axons and terminated on the central region of the intrafusal fibers. Single profiles of terminal axons occupied the sites of sensory neuromuscular junctions, similar to adult sensory endings. No morphological features suggested retraction of afferents from 4-day postnatal spindles. Motor axons approached spindles tightly packed in bundles of 5–20 axons and terminated in the juxtaequatorial and polar regions of both bag and chain fibers. Multiple profiles of terminal axons were visible for each intrafusal motor ending. More motor axons innervated 4-day postnatal spindles and a greater number of axon terminals were visible in immature intrafusal motor endings than in adult spindles. The data suggest that postnatal maturation of motor innervation to intrafusal fibers involves the elimination of supernumerary motor nerve inputs. Synapse elimination in the development of the fusimotor system might represent a mechanism whereby individual axons adjust the number of spindles they innervate.     

Differentiation of myosin in soleus and extensor digitorum longus muscle in different animal species during development

Summary Ca²⁺-ATPase activity and light chains of myosin prepared from fast and slow muscles of rat, guinea-pig and rabbit were studied during development from embryonic to old age to establish further correlation with the well-known developmental changes in contraction properties of these muscles. The changes involve the slow soleus muscle much more than the fast extensor digitorum longus muscle. Myosin-ATPase activity of the soleus muscle before or at birth is higher than in the muscle of adult animals. Myosin from the soleus muscle of embryos or newborn animals reveales light chains of myosin of both fast and slow type (with a preponderance of light chains of fast type in 26-days-old rabbit embryos). During postnatal development the amount of light chains of the fast type decreases, that of the slow type increases. Myosin from the soleus muscle of adult animals contains only light chains of the slow type. However, myosin from the soleus muscle of 30-months-old rats exhibits high myosin ATPase activity and contains light chains of myosin of both slow and fast type as in perinatal development. This is in agreement with the shortening of contraction time observed in this muscle in very old age. Thus developmental differentiation of myosin in the soleus muscle is followed by a trend of levelling out of the differences between fast and slow muscles of senescent animals. No such biphasic development is observed with respect to the fast extensor digitorum longus muscle.Abbreviations EDL extensor digitorum longus - SDS sodium dodecyl sulfate - LC light chain - LC₁ light chain 1     

Distribution patterns of muscle fibre types in major muscles of the bull (Bos taurus).

Summary The study describes the variations in distribution and cross-sectional area (fibre size) of three muscle fibre types (I, IIA, IIB) in 34 of the largest muscles of the bull (Bos taurus). The animals had been kept strictly unexercised for one year before slaughter. Representative sampling was done at 15 positions within each muscle, and from 2700 to 4500 fibres were analysed in each muscle. Different intermuscular patterns are described. The overall volume fraction (%) of type I fibres was about 10% higher in the forepart muscles than in the hindpart muscles (41% and 31%, respectively), while the mean content of type IIB fibres was similar. Type I fibres were particularly abundant in antigravity muscles. Of these, the hindlimb muscles contained 50% more type I fibres (by weight) than those of the forelimb. Typical antigravity antagonists contained very few type I fibres. In the thigh cross-section the proportion of type I fibres was highest in the anterior and medial parts, while the IIB fibres tended to be concentrated in the superficial and posterior parts. Intramuscular patterns were revealed, with type I fibres becoming gradually more abundant from superficial to deep regions, while IIB fibres had an opposite distribution. This was particularly evident in the thigh proper and in the scapular region. Within each fasciculus of all the muscles, the I fibres in the muscles of the forepart were on average about 15% larger than those of the muscles in the hindpart. The IIB fibres were on average about 10% larger in the hindpart than in the forepart muscles. A covariation between the proportion of type I and IIB fibres and their cross-sectional area was indicated.     

Reactive oxygen species and inflammatory mediators enhance muscle spindles mechanosensitivity in rats

We tested the hypothesis that reactive oxygen species (ROS) and inflammatory mediators affect transduction properties of muscle spindles. In rats, muscle spindles response to high-frequency vibration (HFV) was recorded before and after (1) injection of hydrogen peroxide (H2O2) in control rats and animals pre-treated with diclofenac (anti-inflammatory substance), (2) injection of bradykinin and (3) fatigue induced by muscle stimulation (MS) in control rats and rats receiving diclofenac, superoxide dismutase (SOD) or H2O2. Muscular oxidative stress and inflammation induced by H2O2 or MS were assessed by measurements of isoprostanes and IL-6 levels. In control rats, H2O2, bradykinin and MS significantly enhanced the HFV response. Pre-treatment with SOD abolished the post-MS-enhanced HFV response whereas diclofenac lowered the peak HFV response to MS and H2O2. H2O2 injection and MS elicited significant and similar increases in isoprostanes and IL-6. We report a direct modulation of muscle spindles mechanosensitivity by ROS and inflammatory mediators.     

大鼠胸深肌肌纤维型的组成及分布

目的探讨SD大鼠胸深肌的肌纤维型组成和分布,借以了解该肌功能。方法采用Guth-Samaha肌球蛋白ATP酶组织化学染色法并稍做改良,对成年SD大鼠胸深肌冰冻切片进行肌纤维分型研究。结果大鼠胸深肌经肌球蛋白ATP酶组织化学染色后可明确分出2型肌纤维,即明亮色白的Ⅰ型纤维(慢缩纤维)和幽暗深褐的Ⅱ型纤维(快缩纤维),并且,两种纤维在肌内呈棋盘样均匀分布;图像分析仪下计数Ⅱ型纤维达到65%±6%,而Ⅰ纤维仅占35%±5%,前者明显高于后者(P<0.01)。结论大鼠胸深肌以Ⅱ型纤维为主,属于力量和速度型肌。     

Superfluousness of motor innervation for the formation of muscle spindles in neonatal rats

Summary Muscle spindles form de novo in reinnervated muscles of neonatal rats treated with nerve growth factor. Whether the spindles can also form in muscle reinnervated only by afferents was investigated by removing the lumbosacral segment of the spinal cord immediately after crushing the nerve to the medial gastrocnemius muscle at birth, and administering nerve growth factor for 10 days afterwards. As predicted, the medial gastrocnemius muscles were reinnervated by afferents, but not efferents. No motor endplates were visible on any muscle fibers, and extrafusal fibers were atrophied. The reinnervated muscles contained spindle-like encapsulations of one to four fibers at 5, 7, 9 and 30 days after the nerve crush. The number of spindles as well as encapsulated fibers exceeded that of normal medial gastrocnemius muscles. The encapsulated fibers resembled typical intrafusal fibers. They had normal sensory-muscle contacts, but no motor endings. The fibers displayed equatorial clusters of myonuclei and expressed the spindle-specific slow-tonic myosin heavy chain isoform at postnatal day 30. Thus, efferents are not essential for the formation and differentiation of muscle spindles in reinnervated muscles of neonatal rats.     

Exercise-induced fibre type transitions with regard to myosin,parvalbumin, and sarcoplasmic reticulum in muscles of the rat

Effects of a long-term, high intensity training program upon histochemically assessed myofibrillar actomyosin ATPase, myosin composition, peptide pattern of sarcoplasmic reticulum (SR), and parvalbumin content were analysed in muscles from the same rats which were used in a previous study (Green et al. 1983). Following 15 weeks of extreme training, an increase in type I and type IIA fibres and a decrease in type IIB fibres occurred both in plantaris and extensor digitorum longus (EDL) muscles. In the deep portion of vastus lateralis (VLD), there was a pronounced increase from 10±5% to 27±11% in type I fibres. No type I fibres were detected in the superficial portion of vastus lateralis (VLS) both in control and trained animals. An increase in slow type myosin light chains accompanied the histochemically observed fibre type transition in VLD. Changes in the peptide pattern of SR occurred both in VLS and VLD and suggested a complete transition from type IIB to IIA in VLS and from type IIA to I in VLD. A complete type IIA to I transition in the VLD was also suggested by the failure to detect parvalbumin in this muscle after 15 weeks of training. Changes in parvalbumin content and SR tended to precede the transitions in the myosin light chains. Obviously, high intensity endurance training is capable of transforming specific characteristics of muscle fibres beyond the commonly observed changes in the enzyme activity pattern of energy metabolism. The time courses of the various changes which are similar to those in chronic nerve stimulation experiments, indicate that various functional systems of the muscle fibre do not change simultaneously.     

Skeletal fiber types and spindle distribution in limb and jaw muscles of the adult and neonatal opossum,Monodelphis domestica

The South American opossum, Monodelphis domestica, is very immature at birth, and we wished to assess its potential for studies of jaw muscle development. Given the lack of prior information about any Monodelphis fiber types or spindles, our study aimed to identify for the first time fiber types in both adult and neonatal muscles and the location of spindles in the jaw muscles. Fiber types were identified in frozen sections of adult and 6-day-old jaw and limb muscles by using myosin ATPase and metabolic enzyme histochemistry and by immunostaining for myosin isoforms. The distribution of fiber types and muscle spindles throughout the jaw-closer muscles was identified by immunostaining of sections of methacarnoy-fixed, wax-embedded heads. Most muscles contained one slow (type I) and two fast fiber types (equivalent to types IIA and IIX), which were similar to those in eutherian muscle, and an additional (non-IIB) fast type. In jaw-closer muscles, the main extrafusal fiber type was IIM (characteristic of these muscles in some eutherians), and almost all spindles were concentrated in four restricted areas: one in masseter and three in temporalis. Six-day neonatal muscles were very immature, but future spindle-rich areas were revealed by immunostaining and corresponded in position to the adult areas. Extrafusal and spindle fiber types in Monodelphis share many similarities with eutherian mammalian muscle. This finding, along with the immaturity of myosin isoform expression observed 6 days postnatally, indicates that Monodelphis could provide a valuable model for studying early developmental events in the jaw-closer muscles and their spindles. Anat. Rec. 251:548–562, 1998. © 1998 Wiley-Liss, Inc.