Fast Glycolytic Fibers Research Articles

HISTOCHEMICAL PROPERTIES OF MUSCLE ALLOGRAFTS ENHANCED VIA CYCLOSPORINE

Until recently, the transplantation of skeletal muscle across a major histocompatibility barrier has proved difficult. However, with the advent of cyclosporine (CsA), it has become possible to achieve extended survival across such histocompatibility barriers. To date, very little is known about the histochemical, biochemical, immunological or contractile properties of long-term-surviving muscle allografts. Consequently, it was the focus of this study to histochemically examine muscle allografts prolonged with CsA and determine the cross-sectional area of fast glycolytic muscle fibers. Measurements of cross-sectional area were made because they are an important correlate to the amount of tension a muscle can generate. Animals were assigned randomly to one of three groups: control (normal) (n = 5), syngeneic (n = 4), and allogeneic (n = 4). Muscle allografts were performed by transplanting the gastrocnemius of an ACI rat (RT1a) hindlimb into the hindlimb of a Lewis rat (LEW;RT1(1]. The syngeneic model consisted of an ACI-to-ACI transplant. Animals in the allograft group were given CsA (8 mg/kg/day) until the time of sacrifice. At approximately 100 days following transplantation, both syngeneic and allogeneic muscles were removed from the recipient, and quickly frozen in isopentane cooled by liquid nitrogen. Muscle fibers were classified as slow-oxidative (SO), fast-oxidative-glycolytic (FOG), or fast-glycolytic (FG) based upon their staining for myofibrillar ATPase and NADH-dehydrogenase. From each muscle, the cross-sectional area of approximately 175 FG muscle fibers was determined. The fast-glycolytic muscle fibers of both the syngeneic and allogeneic grafts demonstrated a substantial decrease in cross-sectional area. The mean value (+/- SD) for the fibers of the allografted muscle was 1165 +/- 533 microns 2. The mean (+/- SD) fiber cross-sectional area for the fibers of the syngeneic muscle was 973 +/- 421 microns 2. These values contrast with a mean (+/- SD) value of 3552 +/- 601 microns 2 for fibers from age-matched control animals. The differences between the syngeneic and allogeneic muscles were not significant (P greater than 0.05). However, both exhibited significant (P less than 0.01) atrophy compared with the control muscle.

Accumulation of collagen and altered fiber-type ratios as indicators of abnormal muscle gene expression in the mdx dystrophic mouse.

The growth and development of the X-linked muscular dystrophy mutant mouse (mdx) was compared with a control group from 3 weeks to 1 year old. Quantitative cytological analysis of the soleus muscle revealed cycles of degeneration, regeneration, and hypertrophy, and at any one time it was difficult to assess the extent of the disease based on muscle fiber size. One noticeable difference even in the youngest muscles studied was the reduced numbers of slow oxidative fibers and the increased number of fast glycolytic fibers in the mdx soleus muscles. The collagen of the connective tissue components of selectively stained sections was determined by computerized image analysis. Marked accumulation of collagen was found in both the endomysium and perimysium of the dystrophic muscles as compared with age-matched controls. Since the mdx mouse is a result of the same type of genetic defect as in human Duchenne muscular dystrophy, this model could thus be used to assess the effectiveness of various therapeutic approaches, including gene therapy using muscle fibrosis and fiber type proportions as the indicators.

Single muscle fiber enzyme shifts with hindlimb suspension and immobilization.

The purpose of this investigation was to determine how models of weightlessness, hindlimb suspension (HS), and hindlimb immobilization (HI) affect the metabolic enzyme profile in the slow oxidative (SO), fast oxidative glycolytic (FOG), and fast glycolytic (FG) fibers of rat hindlimb. After 1, 2, or 4 wk of HS or HI, single fibers were isolated from freeze-dried soleus and gastrocnemius muscles; a small section of each fiber was run on sodium dodecyl sulfate-polyacrylamide gel electrophoresis gels to identify fiber type, and the remaining piece was assayed for either lactate dehydrogenase (LDH) and citrate synthase (CS) or phosphofructokinase (PFK) and beta-hydroxyacyl-CoA dehydrogenase (beta-OH-acyl-CoA). Two weeks of HS induced an almost twofold increase in the activity of CS (2.13 +/- 0.13 vs. 3.60 +/- 0.26 mol.kg dry wt-1.h-1) in the SO fiber of the soleus, and the activity stayed high at 4 wk. Although the FOG fiber had significantly higher CS activity (3.85 +/- 0.29) than either the SO or FG (1.59 +/- 0.16 mol.kg dry wt-1.h-1) fiber, neither fast fiber type was altered by HS. The glycolytic enzymes LDH and PFK were both elevated in the SO fiber after HS. The increase in LDH occurred by 1 wk (14.80 +/- 1.51 vs. 8.83 +/- 0.78), whereas the activity of PFK was not significantly changed until 4 wk (1.16 +/- 0.13 vs. 0.68 +/- 0.05 mol.kg dry wt-1.h-1). The control FG fiber had the highest LDH (44.30 +/- 2.29) and PFK (2.40 +/- 0.16) activities, followed by the FOG fiber (LDH, 34.10 +/- 2.83; PFK, 1.62 +/- 0.17 mol.kg dry wt-1.h-1); however, the activities of these glycolytic enzymes in the fast fiber types were unaltered by HS. The activity of beta-OH-acyl-CoA was not affected by HS in either the slow or fast fiber types. HI showed qualitatively similar changes to those observed with HS; however, the enzyme shifts developed with a slower time course. In conclusion, both HS and HI shifted the SO fiber enzyme pattern toward that of the control FOG fiber; however, a complete conversion from the SO to FOG fiber did not occur within the 4-wk treatment period.

Effects of postnatal administration of ethanol on the M. gastrocnemius of the albino mouse

A combined morphometric-histochemical (mATPase) study of the effects of ethanol during postnatal development on the m. gastrocnemius has been performed in the albino mouse. The experimental group received ethanol in the drinking water until sacrifice at the age of 40 days. Based on the fiber composition, three different areas are distinguished in the m. gastrocnemius of the mouse. The typical location of these areas does not change after ethanol administration. However, postnatal administration of ethanol produces a selective atrophy and a decrease of the number of type IIb (fast glycolytic) fibers. Concurrently, the number of type IIa (fast oxidative-glycolytic) fibers increases.

Histochemical classification of neck and limb muscle fibers in a turtle, Pseudemys scripta: A study using microphotometry and cluster analysis techniques

We have attempted to develop an objective, semiquantitative classification of fiber types in turtle neck and limb muscle using microphotometry and multivariate statistical techniques. We first stained serial sections for myosin adenosine triphosphatase (ATPase) (with acid and alkaline preincubation and without preincubation), NADH-diaphorase, and two glycolysis-associated markers, alpha-glycerophosphate dehydrogenase (alpha-GPDH) and glycogen phosphorylase A (GPA). This allowed us to characterize individual muscle fibers in terms of their contraction speed and metabolic properties. Next we used microphotometry to measure the optical density of the reaction product in each fiber, and we subjected the resulting optical density matrix to cluster and discriminant function analyses in order to assign fibers to groups (fiber types) and to determine which stains contribute most to the distinction between groups. As a control, we processed a well characterized mammalian muscle (rat sternomastoid) simultaneously. Our results suggest that both neck and limb muscle in Pseudemys can best be described as falling into three groups: 1) slow oxidative (SO) fibers; 2) fast oxidative glycolytic (FOG) fibers, with relatively high oxidative and glycolytic capacities; and 3) fast glycolytic (Fg) fibers, with low oxidative, low/intermediate alpha-GPDH, and high GPA activities. These three fiber types differ from like-named types in rat muscle both in the pH lability of their myosins and in their metabolic profiles.

Fibre typing using sarcoplasmic reticulum Ca2+-ATPase and myoglobin immunohistochemistry in rat gastrocnemius muscle

Skeletal muscle fibre types were identified by using immunohistochemical detection of sarcoplasmic reticulum Ca2+-ATPase and myoglobin content in rat gastrocnemius muscle. The strong Ca2+-ATPase-reactive fibres were identical with the fast-twitch population, while the fibres with weak reactivity represented the slow-twitch type. Strong myoglobin immunoreactivity reflected the fast oxidative glycolytic (FOG) and slow oxidative (SO) types. Slight to moderate myoglobin immunostaining was found in the fast glycolytic (FG) fibres. The staining intensity of the different fibre types differed as follows: for Ca2+-ATPase FG greater than FOG greater than SO, and for myoglobin FOG greater than SO greater than FG. The immunoreactivity of Ca2+-ATPase and myoglobin were well preserved after fixation of the muscles in Bouin's solution, or in formol/acetic acid fixative, and paraffin embedding. Detection of the primary antibodies was carried out by using the avidin-biotin-peroxidase complex, and the immunogold-silver-staining methods. The latter was found to be more sensitive and suitable for postembedding ultrastructural demonstration of the Ca2+-pump enzyme on Durcupan-embedded muscles. The method, using 5 nm immunogold conjugate with silver enhancement, offered the advantages of high sensitivity and excellent visualization of the reaction product. The postembedding detection of sarcoplasmic reticulum Ca2+-ATPase also proved to be useful in the retrospective identification of the main fibre classes in human muscle biopsies.

Structure and Function in Vertebrate Skeletal Muscle

its structure. An important aspect of this is its hierarchical design. At the cellular level, muscle fibers form three categories whose functional properties grade into each other: slow-oxidative fibers with high endurance to fatigue, fast-oxidative/glycolytic fibers also endurant but with greater metabolic diversity, and fast-glycolytic fibers with limited endur? ance but quick response. This partitioning of functional properties found among single muscle fibers is conserved at a second level of the structural hierarchy, since the group of myofibers innervated by a single motor neuron (together called a motor unit) is com? posed ofthe same fiber type. Different motor units may be recruited in an orderly pattern depending upon the functional demands ofa particular behavior. Finally, groups of motor units innervated by axons travelling together in the primary nerve branches may form discrete neuromuscular compartments at a third level of structural hierarchy. Different motor units may be found in regional arrays in these compartments so that slow or fast units tend to be clumped together and may be recruited together as larger functional units. This hierarchical organization of skeletal muscle may be a fundamental vertebrate plan that allows the diversity of functions so evident in vertebrate behavior.

Influence of fiber type and muscle source on Ca2+ sensitivity of rat fibers

This study investigated the influence of muscle source and fiber type on the calcium sensitivity of skinned rat skeletal muscle fibers from predominantly slow muscles [soleus (SOL) and adductor longus (AL)], mixed muscle [posterior gracilis (PG)], and predominantly fast-twitch muscle [extensor digitorum longus (EDL)]. Fibers were characterized histochemically and by one-dimensional protein gel electrophoresis, and calcium-tension relationships were determined. Fiber type and muscle source had significant effects on the negative log of the calcium concentration associated with half-maximal tension (pCa1/2). Slow-twitch fibers had larger values of pCa1/2 than did fast-twitch fibers. Slow-twitch fibers from the predominantly slow muscles, SOL and AL, had similar values of pCa1/2 but slightly smaller values than from the mixed muscle, PG. Fast-glycolytic (FG) fibers from the predominantly fast muscle, EDL, had a higher pCa1/2 than fibers from the mixed fiber type muscle, PG. There were no differences between the pCa1/2 associated with FG and fast-oxidative-glycolytic fibers.

EFFECTS OF LONG‐TERM STREPTOZOTOCIN DIABETES ON THE CONTRACTILE AND HISTOCHEMICAL PROPERTIES OF RAT MUSCLES

Contractile and histochemical properties of soleus (a slow-twitch muscle) and extensor digitorum longus (EDL, a fast-twitch muscle) were studied in mature rats after 3 months of streptozotocin-induced diabetes. Results were compared with age- and weight-matched controls. Diabetes produced profound wasting of fast muscles and particularly of the fast glycolytic (FG) fibres. Slow muscle fibres, both within the mixed EDL and in soleus, were less atrophied. Strength performance of EDL was reduced by diabetes, but maintained in soleus. Diabetes was without effect on the time to peak tension (TTP) and half-relaxation time (HRT) of EDL. However it produced profound slowing of soleus muscles, particularly of the relaxation phase. Part of the slowing effect of diabetes may be related to a histochemically demonstrable loss of fast oxidative glycolytic (FOG) fibres in soleus. Histochemical staining for the oxidative marker succinic dehydrogenase (SDH) revealed marked disruption of reaction product distribution in soleus, indicating an impairment of oxidative capacity.

Calcium sensitivity of fast‐and slow‐twitch human muscle fibers

Muscle fibers from the lateral gastrocnemius muscles of five normal adult males were chemically skinned (sarcolemma disrupted) and each fiber was divided into two parts for histochemical determination of fiber type and calcium-induced tension measurements. The calcium concentration associated with half-maximal tension was 2.5 microM for fast-twitch and 1.3-1.4 microM for slow-twitch fibers. Fast-oxidative-glycolytic (type 2A) and fast-glycolytic (type 2B) fibers had similar calcium sensitivities. Maximum tensions (kg/cm2) were 2.24 for fast-twitch and 2.19 for slow-twitch fibers.

In situ hybridization and immunocytochemistry in serial sections of rabbit skeletal muscle to detect myosin expression.

We performed in situ hybridization of myosin heavy-chain (MHC) mRNA on rabbit muscle using a biotin-labeled complementary RNA probe. An 1107-nucleotide fragment from an alpha-cardiac MHC cDNA was used to transcribe an RNA probe 97% similar to slow-twitch and 75% similar to fast-twitch sequences. Serial sections were used to identify slow-twitch fibers in medial gastrocnemius, soleus, and tibialis anterior by immunofluorescence of slow MHC and oxidative capacity by histochemistry. Slow-twitch fibers hybridized by the RNA probe stained heavily after detection with streptavidin-alkaline phosphatase (89% dark and 11% medium density). Fast-oxidative fibers stained intermediately (26% dark, 58% medium, and 16% light) and fast-glycolytic fibers stained lightly (12% medium and 88% light). Biotin-labeled probe and enzymatic detection allowed greater resolution of the subcellular location of the MHC mRNA, a distinct advantage over isotope labeling and autoradiography. A non-uniform distribution of MHC mRNA was recognized within an adult skeletal muscle fiber. High concentrations of MHC mRNA were found under the sarcolemma and between the myofibrils, suggesting the existence of a distribution mechanism. The combination of in situ hybridization and immunocytochemistry allows rapid subcellular localization of both MHC mRNA and its translated protein.

Selective damage of fast glycolytic muscle fibres with eccentric contraction of the rabbit tibialis anterior.

Acta Physiologica ScandinavicaVolume 133, Issue 4 p. 587-588 Selective damage of fast glycolytic muscle fibres with eccentric contraction of the rabbit tibialis anterior R. L. LIEBER, R. L. LIEBER Division of Orthopaedics and Rehabilitation, Veterans Administration Medical Center and University of California, San Diego, USASearch for more papers by this authorJ. FRIDÉN, J. FRIDÉN Department of Hand and Reconstructive Surgery, Umeå University Hospital, S-901 85 Umeå, SwedenSearch for more papers by this author R. L. LIEBER, R. L. LIEBER Division of Orthopaedics and Rehabilitation, Veterans Administration Medical Center and University of California, San Diego, USASearch for more papers by this authorJ. FRIDÉN, J. FRIDÉN Department of Hand and Reconstructive Surgery, Umeå University Hospital, S-901 85 Umeå, SwedenSearch for more papers by this author First published: August 1988 https://doi.org/10.1111/j.1748-1716.1988.tb08446.xCitations: 141AboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onFacebookTwitterLinked InRedditWechat No abstract is available for this article.Citing Literature Volume133, Issue4August 1988Pages 587-588 RelatedInformation

Glycogen utilization in rat respiratory muscles during intense running.

Glycogen concentration in the adult rat diaphragm and intercostal muscles has been examined following heavy treadmill exercise to determine the recruitment strategy and the significance of glycogen as a substrate to satisfy the elevated energy requirements accompanying hyperpnea. Short-term continuous running at 60 m/min and a 12 degree grade resulted in a reduction (p less than 0.05) in the concentration of glycogen (39%) in the costal region of the rat diaphragm. Similarly, glycogen concentration was significantly reduced (p less than 0.05) with this exercise protocol in all respiratory muscles studied, with the exception of the sternal region of the diaphragm. With the less intense running protocols, glycogen degradation continued to be pronounced (p less than 0.05) in the majority of the respiratory muscles sampled. The significance of muscle glycogen as a substrate for energy metabolism in the respiratory muscles was not affected by the procedure used to prepare the animal for tissue sampling (Somnitol, diethyl ether, decapitation). Examination of selected locomotor muscles revealed extensive glycogen loss in muscles composed of essentially slow oxidative fibres (soleus), fast oxidative glycolytic fibres (vastus lateralis red), and fast glycolytic fibres (vastus lateralis white). It is concluded that during heavy exercise in the rat, recruitment of motor units occurs in all regions of the diaphragm and in the intercostal muscles. At least for the costal region of the diaphragm and as evidenced by the modest (two- to four-fold) but significant (p less than 0.05) increases in lactate concentration, the increased ATP requirements in these muscles are met to a large degree by increases in aerobic metabolism.(ABSTRACT TRUNCATED AT 250 WORDS)

Comparison of physiological and histochemical properties of motor units after cross-reinnervation of antagonistic muscles in the cat hindlimb.

1. Physiological and histochemical properties of the cat ankle extensor muscles, the lateral and medial gastrocnemius, and the soleus were studied after cross-reinnervation by flexor motoneurons. 2. Tibial and common peroneal nerves were cut and cross-united in the popliteal fossa of 2- to 6-mo-old cats. Eighteen to 24 mo later, single motor units were isolated by dissection and stimulation of ventral root filaments and classified into four types: fast-twitch, fatigable (FF), fast-twitch with intermediate fatigue resistance (FI), fast-twitch, fatigue-resistant (FR), and slow, fatigue-resistant (S). Muscle fibers were classified as fast glycolytic (FG), fast, oxidative glycolytic (FOG), and slow oxidative (SO) on the basis of histochemical staining. 3. Although motor-unit force was normally well correlated with the size of the innervating motor axon in the cross-reinnervated muscles, the force of different unit types overlapped considerably. The reinnervated motor units also showed a higher than normal degree of fatigability. 4. The range of muscle unit forces in cross-reinnervated triceps surae muscles was the same as in the normally innervated triceps surae muscles. This range is 2-3 times greater than the flexor muscles, which the common peroneal nerve normally supplies. The range of contraction speed of units in the cross-reinnervated extensor muscles was comparable to that in the flexor muscles, consistent with a motoneuron-specific determination of muscle speed (28). 5. SO and FOG muscle fibers were found in all reinnervated triceps surae muscles, but FG fibers were only found in reinnervated medial gastrocnemius (MG) and lateral gastrocnemius (LG) muscles, consistent with previous findings of the resistance of soleus muscles to complete conversion (10, 16, 20, 21). Type grouping of muscle fibers was characteristic of the reinnervated muscles. 6. Reinnervated SO muscle fibers were larger than the corresponding fibers in normally innervated muscles as were the estimated number of muscle fibers innervated by slow motor axons. Nonetheless, the force generated by the S motor units remained relatively smaller than FR and FF units. The relative contributions of the number, cross-sectional area and specific tension to the force generation of reinnervated motor units are discussed.

Comparative aspects of fibre types, areas, and capillary supply in the pectoralis muscle of some passerine birds with differing migratory behaviour

Fibre types, fibre areas and capillary supply in the pectoralis muscle of fifteen passerines with four different patterns of migratory behaviour have been studied. The predominant fibre type was a fast-twitch oxidative-glycolytic which was the only fibre type present in all species, except in the robin and the blackbird where a fast fibre with intermediate oxidative capacity and a fast glycolytic fibre were also found. There was a significant difference in fibre areas between birds with different migratory strategies, with the long-distance migratory group having the smallest fibres. This also led to higher capillary densities, shorter diffusion distances and, consequently, more capillaries around the fibres relative to fibre area in this group. This indicates an adaptation in the morphology of the pectoralis muscle to differences in migration strategies. In the robin, the proportion of the intermediate fibre was significantly greater during the breeding season than during migration. Seasonal differences in fibre areas and capillary supply within a species were also seen, but no definite trends were detectable.

The distribution of anterogradely labeled I--IV primary afferents in histochemically defined compartments of the rat's sternomastoid muscle.

The sternomastoid muscle of the rat is divided into a white (dominated by fast-glycolytic twitch fibers) and a red (dominated by fast oxidative-glycolytic twitch fibers, but also containing slow-oxidative twitch fibers) compartment. Previous reports on exclusive location of muscle spindles in the red portion were confirmed. On the basis of anterograde labeling with horseradish peroxidase-wheat germ agglutinine conjugate (WGA-HRP) it was shown in this study that, in addition to muscle spindle compartmentalisation, there was also an exclusive occurrence of tendon organs in the red part of the muscle; moreover, fine afferents (III- and IV-afferents) were mainly distributed to this portion as well. Radioimmunassay studies revealed that this part of the muscle contained twice as much substance P as the white part. It could be shown by acetylcholinesterase (AChE) histochemistry that the myelinated fibers of the white branch to the muscle exclusively displayed high enzyme activity which is characteristic for motor fibers; on the other hand, in the branch to the red portion two classes of AChE-positive fibers were found: a large one with a peak in the alpha-range, and a small one with a peak in the gamma-range. In addition, there was also a group of enzyme-negative (sensory) fibers. These results also indicate the red portion of the sternomastoid muscle to be its "sensory compartment".

Structure of abdominal muscles in the hamster: effect of elastase-induced emphysema.

The present study examined the effects of elastase-induced emphysema on the structure of the external oblique and transverse abdominis muscles and a non-respiratory muscle, the extensor digitorum longus. Muscle structure was assessed from the cross-sectional area (CSA) and percent of individual fiber types in histochemically stained sections and from the number of sarcomeres arranged in series along the length of individual fibers. Data were obtained in eight hamsters with emphysema and nine saline-injected controls. In the normal (control) animals the external oblique was thicker but contained fewer sarcomeres than the transverse abdominis. Fiber size was similar in the two muscles. In the transverse abdominis the percents of fast-glycolytic and fast-oxidative fibers were greater and smaller, respectively, than in the external oblique. Lung volume of emphysematous hamsters was 168% of control values (P less than 0.001). In emphysematous compared with control animals, the CSA of fast-twitch fibers in the external oblique and transverse abdominis was significantly reduced. Fiber length and sarcomere number were significantly decreased in the transverse abdominis but not in the external oblique in emphysematous hamsters. In contrast, fiber size and composition of the extensor digitorum longus was similar in emphysematous and control animals. These data indicate that cellular responses of the ventilatory muscles to chronic hyperinflation and altered thoracic geometry induced by emphysema are not present in limb skeletal muscle. We speculate that changes in fiber length and CSA of fast fibers in the abdominal expiratory muscles reflect responses to chronic alterations in the mechanics of breathing that may affect muscle load, length, or the pattern of activity.

Fiber-specific cholesterol changes in murine dystrophy

The cholesterol concentration in dystrophic mouse muscle is reported to be increased compared to normal. The muscles studied are, however, composed in most cases of more than one fiber type. As a result, the observed concentration increase may be due to a general increase or may be due to changes in the proportion of individual fiber types which themselves differ in cholesterol concentration. To decide between these possibilities we have measured the cholesterol concentrations (both free cholesterol and cholesterol esters) in normal and dystrophic whole gastrocnemius muscles and compared the values with the concentrations in fast-glycolytic muscle tissue alone. The cholesterol concentrations in both whole and fast-glycolytic sections of dystrophic muscle are increased compared to normal, with the largest increase in the cholesterol ester fraction. Furthermore, the concentration changes in fast-glycolytic fibers are due mainly to cholesterol ester differences in both membrane and sarcoplasm fractions, with differences in the latter being larger. The data show that changes in whole muscle concentrations cannot be ascribed solely to altered fiber type proportions.

Fiber types in normal and neonatally denervated fast muscles of the rat: Immunocytochemical study with an antimyosin monoclonal antibody

The differentiation of fiber types in normal and neonatally denervated gastrocnemius muscles of the rat was compared by myosin ATPase histochemistry and immunocytochemistry using a monoclonal antibody, HM-1.2. The specificity of HM-1.2 for the fast myosin heavy chain was determined by radioimmunoassay, immunoautoradiography, and indirect immunofluorescence techniques. In normal 1-month-old and adult rats, the type IIB (fast glycolytic) fibers of the gastrocnemius could be clearly divided into three subtypes by their graded immunofluorescence staining with the myosin heavy chain-specific monoclonal antibody. In the gastrocnemius muscle of the newborn rat, all fibers were negative with the monoclonal antibody. The transition from negative to three grades of immunoreactivity occurred 1 to 2 weeks postnatally. After neonatal denervation of the gastrocnemius muscle, however, uniformly positive monoclonal antibody immunofluorescence staining for the myosin heavy chain was observed without subtype differentiation. This study, thus, gave clear immunocytochemical evidence that the type IIB muscle fibers are heterogeneous with respect to their myosin isoform and that the expression of this heterogeneity is dependent on the normal developmental influence of motor innervation on the muscle fibers.

Morphometry, histochemistry, and contractility of dystrophic hamster diaphragm

Differences in gross and microscopic morphology, fiber-size distribution, and fiber-type composition were present in the diaphragm of 35-, 130-, and 180-day-old dystrophic (Bio 14.6) compared with age-matched control (Bio F1B) hamsters. The dystrophic diaphragm was significantly thicker than the control at 130 and 180 days. Increases in wet-to-dry weight ratios, connective tissue per unit area, and muscle fiber number suggest that increased tissue hydration, fibrosis, and fiber hyperplasia contribute to diaphragm hypertrophy. Marked variations of fiber areas and diameters were evident in each fiber type at each age, but generalized atrophy predominated over hypertrophy, resulting in significant decreases in cross-sectional areas of each fiber type. Significant differences in fiber-type composition were noted in the dystrophic vs. control diaphragm at each age: at 35 days the percentage of slow-oxidative fibers was lower and in fast-oxidative fibers was higher; at 130 days the percentage of fast-oxidative fibers remained elevated; at 180 days the percentage of fast-glycolytic fibers was reduced. In vitro contractility studies of 130-day-old animals showed that twitch and peak tension development were significantly lower in the dystrophic compared with the control diaphragm, whereas optimal length, contraction time, and half-relaxation time were within control limits. Microscopic and physiological abnormalities were also present in the soleus of 130-day-old dystrophic animals. As in the diaphragm, fiber areas were reduced, connective tissue area increased, and peak and twitch tension decreased significantly compared with the control soleus. The histopathological and pathophysiological changes in the diaphragm correlated well with each other and are consistent with the slowly evolving inability of the dystrophic hamster to increase tidal volume and minute ventilation in response to a hypercapnic challenge.