2b Fibers Research Articles

Fiber-type specific and position-dependent expression of a transgene in limb muscles

We have previously shown that the proximal sequences of the human aldolase A fast-muscle-specific promoter (pM) are sufficient to target the expression of a linked CAT reporter gene to all fast, glycolytic trunk and limb muscles of transgenic mice (pM310CAT lines) in a manner mimicking the activity of the endogenous mouse promoter. When a NF1-binding site (motif M2) in this proximal regulatory region is mutated, the activity of the corresponding mM2 transgene is strongly affected but only in a some fast muscles. Here we show that the mutation of the M2 motif has only mild effects on pM activity in axial and proximal limb, while it drastically reduces this activity in both fore and hind limb distal muscles. At the cellular level, we show that both the pM310CAT and mM2 transgenes are highly expressed in fast glycolytic 2B fibers. However, by contrast to the pM310CAT transgene, whose expression is mainly restricted to fast glycolytic 2B fibers, the mM2 transgene is also active in a high proportion of 2X fibers. This result suggests that the M2 sequence could play a role in restricting the expression of pM to the 2B fibers. The variable expression of the mM2 transgene along the limb axis already exists at post-natal day 10 and seems to result from a change in the proportion of expressing fast fibers per muscle. Altogether, these results suggest that, although considered as phenotypically similar, different populations of fast glycolytic fibers exist, in which the requirement of the NF1 activity for pM expression varies according to the proximal versus distal position of the muscle along the limb axis.

Relative distribution of three major lactate transporters in frozen human tissues and their localization in unfixed skeletal muscle.

We have prepared affinity-purified rabbit polyclonal antibodies to the near-C-terminal peptides of human monocarboxylate transporters (MCTs) 1, 2, and 4 coupled to keyhole limpet hemocyanin. Each antiserum reacted only with its specific peptide antigen and gave a distinct molecular weight band (blocked by preincubation with antigen) after chemiluminescence reaction on Western blots from sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) of tissue membrane proteins. Densitometry showed distinctive expression patterns for each MCT in a panel of 15 frozen human tissues, with the distribution of MCT1 >>MCT2>MCT4. Fluorescence microscopy of unfixed skeletal muscle using fluorescein-conjugated secondary antibody was correlated with reverse adenosine triphosphatase (ATPase) stained sequential sections to identify fiber-type localization. MCT1 expression was high in the sarcolemma of type 1 fibers, modest to low in type 2a fibers, and almost absent in type 2b fibers. In contrast, MCT4 expression was low to absent in the membrane of most type 1 fibers, but high in most 2a and in all 2b fibers, favoring the view that their high lactate levels during work may be channeled in part to neighboring type 1 (and perhaps 2a) fibers for oxidation, thereby delaying fatigue. MCT2 expression was limited to the sarcolemma of a type 1 fiber subset, which varied from <5 to 40%, depending on the specific muscle under study. Quantitative chemiluminescent densitometry of 10 muscle biopsies for their MCT2 and MCT4 content, each normalized to MCT1, confirmed the unique variation of MCT2 expression with biopsy site. The application of these antibodies should add to the understanding of motor unit physiology, and may contribute to the muscle-biopsy assessment of low-level denervation.

Fibre type composition of soleus and extensor digitorum longus muscles in normal female inbred Lewis rats

We have analysed the fibre type composition of soleus and extensor digitorum longus (EDL) muscles of normal female 4-6-month-old inbred Lewis rats. This rat strain is used in our ongoing study of the effects of thyroid hormone on myosin heavy chain (MyHC) isoform expression. On the basis of the mATPase reaction, soleus muscles contained 96.1 +/- 2.9% of type 1 fibres supplemented by 2A fibres. EDL muscles contained type 1 (5.5 +/- 1.0%), type 2A (18.8 +/- 1.7%) and type 2B (75.7 +/- 2.2%) fibres. Immunohistochemical analysis and SDS gel electrophoresis confirmed that most fibres in the soleus muscle expressed the type 1 (slow) MyHC isoform and that only a small proportion of fibres expressed the fast 2a MyHC isoform. Immunohistochemical analysis and SDS gel electrophoresis demonstrated that almost half of the 2B fibres of EDL muscles expressed the 2x/d MyHC isoform. In both muscle types, many fibres expressed more than one MyHC isoform. The content of slow fibres in the soleus muscle of female inbred Lewis rats was slightly higher than that reported for Wistar rats, but was considerably higher than that of Sprague-Dawley rats, whereas substantial differences were not found in the proportion of slow and fast fibre types in EDL muscles in these strains.

Myostatin, insulin-like growth factor-1, and leukemia inhibitory factor mRNAs are upregulated in chronic human disuse muscle atrophy.

Human disuse muscle atrophy frequently accompanies orthopedic injury, arthritis, or bed rest, and recovery is often incomplete despite current rehabilitation programs. We have studied the vastus lateralis muscle in 12 patients with chronic disuse atrophy associated with chronic osteoarthritis of the hip both preoperatively and after total hip arthroplasty. Semiquantitative reverse transcriptase-polymerase chain reaction (RT-PCR) demonstrated an increase in the level of expression of myostatin, insulin-like growth factor-1 (IGF-1) and leukemia inhibitory factor (LIF) mRNAs compared to healthy control muscle. In all patients there was a significant correlation preoperatively between increasing myostatin mRNA expression and reduction in type 2A and 2B fiber area. In the 8 female patients there was a significant correlation between increased myostatin mRNA expression and the atrophy factor calculated for 2A and 2B fiber types preoperatively. We hypothesize that a complex interaction occurs between muscle growth regulating factors in the genesis of muscle wasting. Our results indicate that myostatin is a muscle-wasting factor contributing to type 2B and 2A atrophy. Other muscle growth factors, such as IGF-1 and LIF, may be upregulated in a counterregulatory fashion or may be involved in the fiber type switching seen in disuse muscle wasting.

Nutritional strategy in the management of heart failure in adults.

The incidence of congestive heart failure (CHF) is increasing in Westernized countries, and patients with CHF experience poor quality of life (functional impairment, high hospitalization rate and high mortality). Malnutrition occurring during the course of CHF is referred to as cardiac cachexia and is associated with higher mortality independent of the severity of CHF. Cardiac cachexia involving a loss of more than 10% of lean body mass can clinically be defined as a bodyweight loss of 7.5% of previous dry bodyweight in a period longer than 6 months. The energy requirements of patients with CHF, whether cachectic or not, are not noticeably modified since the increase in resting energy expenditure is compensated by a decrease in physical activity energy expenditure. Malnutrition in CHF has been ascribed to neurohormonal alterations, i.e. anabolic/catabolic imbalance and increased cytokine release. Anorexia may occur, particularly during acute decompensation of CHF. Function is impaired in CHF, because of exertional dyspnea and changes in skeletal muscle. Decreased exercise endurance seems to be related to decreased mitochondrial oxidative capacities and atrophy of type 1 fibers, which are attributed to alteration in muscle perfusion and are partially reversible by training. Malnutrition could also impair muscle function, because of decreased muscle mass and strength associated with decreased glycolytic capacities and atrophy of type 2a and 2b fibres. With respect to the putative mechanisms of cardiac cachexia, anabolic therapy (hormones or nutrients) and anticytokine therapy have been proposed, but trials are scarce and often inconclusive. In surgical patients with CHF, perioperative (pre- and postoperative) nutritional support has been shown to be effective in reducing the mortality rate. Long term nutritional supplementation trials in patients with CHF and cachexia are thus required to establish recommendations for the nutritional management of patients with CHF.

Expression of the Na(+)/Ca(2+) exchanger in skeletal muscle.

The expression of the Na(+)/Ca(2+) exchanger was studied in differentiating muscle fibers in rats. NCX1 and NCX3 isoform (Na(+)/Ca(2+) exchanger isoform) expression was found to be developmentally regulated. NCX1 mRNA and protein levels peaked shortly after birth. Conversely, NCX3 isoform expression was very low in muscles of newborn rats but increased dramatically during the first 2 wk of postnatal life. Immunocytochemical analysis showed that NCX1 was uniformly distributed along the sarcolemmal membrane of undifferentiated rat muscle fibers but formed clusters in T-tubular membranes and sarcolemma of adult muscle. NCX3 appeared to be more uniformly distributed along the sarcolemma and inside myoplasm. In the adult, NCX1 was predominantly expressed in oxidative (type 1 and 2A) fibers of both slow- and fast-twitch muscles, whereas NCX3 was highly expressed in fast glycolytic (2B) fibers. NCX2 was expressed in rat brain but not in skeletal muscle. Developmental changes in NCX1 and NCX3 as well as the distribution of these isoforms at the cellular level and in different fiber types suggest that they may have different physiological roles.

Monospecific Antibodies against the Three Mammalian Fast Limb Myosin Heavy Chains

Skeletal muscle fibres in mammalian limb muscles are of four types: slow, 2A, 2X, and 2B, each characterized by a distinct myosin heavy chain (MyHC) isoform. Existing monoclonal antibodies (mabs) against fast MyHCs lack fibre-type specificity across species and could not positively identify 2X fibres. In this work, mabs were raised against each of the fast MyHCs. These mabs were shown to be monospecific by Western blots and immunohistochemistry in the rat. The advantages of using these mabs for identifying the three fast fibre types and hybrid fibres expressing multiple isoforms were illustrated using rat tibialis anterior muscle. Immunohistochemical analyses confirmed the monospecificity of these mabs in the following additional species: mouse, guinea pig, rabbit, cat, and baboon. 2B fibres were absent in limb muscles of the cat and baboon. These mabs constitute a set of powerful tools for studying muscle fibre types and their transformations.

Specific contributions of various muscle fibre types to human muscle performance: an in vitro study

Human skeletal muscle fibres can be divided in five groups: 1, 1–2A, 2A, 2A–2B and 2B, by using myosin heavy chain (MHC) isoforms as molecular markers. This study aimed to define the contribution of each fibre type to the contractile performance of human muscles. Single fibre segments were dissected from bioptic samples of vastus lateralis and chemically skinned. Force–velocity properties, including isometric tension ( P 0), maximal shortening velocity ( V max), maximum power output ( W max) and the velocity at which W max is reached ( V opt), were determined at maximum calcium activation. Among these parameters W max showed the largest range of variation: about nine times between 2B and slow fibres. V opt also showed large (about four times) and significant variations between fibre types. Force development at submaximum calcium activation was studied and force–pCa curves were obtained for each fibre type. Calcium sensitivity was greater in 2B than in 2A and in slow fibres. The slope of the force–pCa curve was greater in fast than in slow fibres. At the end of the experiment the MHC isoform composition of each fibre segment was determined by gel electrophoresis. The functional properties of each fibre type are discussed in the light of the motor unit recruitment mechanism to understand their possible physiological role.

Early Decrease of IIx Myosin Heavy Chain Transcripts in Duchenne Muscular Dystrophy

We studied byin situhybridization the expression of IIx myosin heavy chain (MHC) transcripts in Duchenne Muscular Dystrophy (DMD) muscle. In normal muscle fibers IIx MHC transcripts were only expressed in type 2b or very fast fibers. Our results indicate that muscle fibers expressing IIx MHC transcripts disappear early in DMD muscle. This may be due to the transformation of type 2b fibers into type 2a fibers and/or to selective degeneration of the type 2b fibers. The very fast 2b fibers may be subjected to a greater mechanical stress during muscle contraction and dystrophin deficiency may render them more susceptible to mechanical damage.

Proximal Sequences of the Aldolase A Fast Muscle-specific Promoter Direct Nerve- and Activity-dependent Expression in Transgenic Mice

Muscle activity is known to modulate the muscle fiber phenotype. Changes in muscle activity (normal or experimentally induced) lead to modifications of the expression status of several muscle-specific genes. However, the transcription regulatory elements involved in the adaptative response are mainly unknown. The aldolase A muscle-specific promoter, pM, is expressed in adult fast twitch muscle with a preferential expression in fast glycolytic-2B fibers. Its activity is induced during postnatal muscle maturation, suggesting a role of nerve and/or muscle activity. Indeed, denervation of gastrocnemius in newborn mice prevented the activation of the promoter in this muscle, despite the nerve-independent formation of 2B fibers. Although the nerve was necessary for pM onset during development, denervating the gastrocnemius in adults had only mild effects on pM activity. By contrast, a transgene including the pM proximal regulatory sequences that are sufficient to reproduce the 2B fiber-specific expression of the endogenous promoter was shown to be highly sensitive to both neonatal and adult denervation. Transgenes containing muscle-specific pM proximal promoter elements were used to delineate the regulatory elements involved in this response to innervation and changes in the contractile activity pattern. Nerve- and activity-dependent elements could be localized in the 130-base pair-long proximal promoter region of the human aldolase A gene.

EFFECT OF ACTIVITIES OF DAILY LIVING ON FIBER TYPE ATROPHY OF THE VASTUS LATERALIS MUSCLE IN PATIENTS WITH JOINT DISORDERS

The histopathologic study was performed to elucidate whether the fiber type atrophy of the vastus lateralis muscle in patients with hip or knee joint disorders is related to the activities of daily living (ADL) or habitual physical activity. Subjects were 16 female patients, 52.4 +/- 16.0 yr of age (mean +/- standard deviation), who underwent a vastus lateralis muscle biopsy at the time of total hip or knee replacement. At the time of referral to the rehabilitation center, the Functional Independence Measure (FIM) motor score and habitual physical activity at home were evaluated, and the diameter and atrophy factor for each muscle fiber type were measured on the histopathologic preparations of the biopsied muscles. The data were analyzed using ttest, one-way analysis of variance (ANOVA), Kruskal-Wallis one-way ANOVA, Spearman's correlation coefficient, and partial correlation coefficient. The patients showed muscle fiber atrophy and small angular fibers, and the atrophy factor was significantly increased in type 1, 2A, and 2B fibers, in that order (one-way ANOVA, P < 0.05). The patterns of the fiber type atrophy, consisting of normal, type 2B atrophy, type 2AB atrophy, and type 1 and 2AB atrophy, had a significant relationship with the fiber type atrophy (Spearman's correlation coefficient; rho = 0.834, P < 0.001). The FIM motor score showed a significant correlation with the atrophy factor (r = -0.584, P < 0.05), and significant differences were recognized among the four patterns of the fiber type atrophy (Kruskal-Wallis one-way ANOVA, P < 0.05). In conclusion, the muscle atrophy and patterns of the fiber type atrophy of the vastus lateralis muscle in patients with joint disorders may be related to changes in the FIM motor score.

Neurotrophin-3-enhanced nerve regeneration selectively improves recovery of muscle fibers expressing myosin heavy chains 2b.

The purpose of this study was to evaluate the effect of neurotrophin 3 (NT-3) enhanced nerve regeneration on the reinnervation of a target muscle. Muscle fibers can be classified according to their mechanical properties and myosin heavy chain (MHC) isoform composition. MHC1 containing slow-type and MHC2a or 2b fast-type fibers are normally distributed in a mosaic pattern, their phenotype dictated by motor innervation. After denervation, all fibers switch to fast-type MHC2b expression and also undergo atrophy resulting in loss of muscle mass. After regeneration, discrimination between fast and slow fibers returns, but the distribution and fiber size change according to the level of reinnervation. In this study, rat gastrocnemius muscles (ipsilateral and contralateral to the side of nerve injury) were collected up to 8 mo after nerve repair, with or without local delivery of NT-3. The phenotype changes of MHC1, 2a, and 2b were analyzed by immunohistochemistry, and fiber type proportion, diameter, and grouping were assessed by computerized image analysis. At 8 mo, the local delivery of NT-3 resulted in significant improvement in gastrocnemius muscle weight compared with controls (NT-3 group 47%, controls 39% weight of contralateral normal muscle; P < 0.05). NT-3 delivery resulted in a significant increase in the proportion (NT-3 43.3%, controls 35.7%; P < 0.05) and diameter (NT-3 87.8 micron, controls 70.8 micron; P < 0.05) of fast type 2b fibers after reinnervation. This effect was specific to type 2b fibers; no normalization was seen in other fiber types. This study indicates that NT-3-enhanced axonal regeneration has a beneficial effect on the motor target organ. Also, NT-3 may be specifically affecting a subset of motoneurons that determine type 2b muscle fiber phenotype. As NT-3 was topically applied to cut nerves, our data suggest a discriminating effect of the neurotrophin on neuro-muscular interaction. These results would imply that muscle fibers may be differentially responsive to other neurotrophic factors and indicate the potential clinical role of NT-3 in the prevention of muscle atrophy after nerve injury.

Energy metabolism of fibre types within fascicles of human muscles

In human latissimus dorsi muscle a preponderance of type 2b fibres in the first fascicle layer and of type 1 fibres in the second layer was found. NADH-dehydrogenase (NADH) and alpha-glycerophosphate dehydrogenase (GPDH) which were measured histophotometrically in type 1, 2a, and 2b fibres showed either extreme or only partial overlapping regarding the activity of metabolic enzymes. In different fascicle layers the average activity of both enzymes did not differ significantly among the fibres of the same type, neither did the NADH and GPDH activity of type 2a and 2b.

Inherited muscular disorder in mutant Japanese quail ( Coturnix coturnix japonica): Relationship between the development of muscle lesions and age

The progression of the pathological changes that occur in the skeletal muscle was examined in 19 Japanese quail of the LWC strain, affected with an autosomal dominant inherited muscular disorder producing electrical myotonia. The muscle samples were obtained every 10 days from 20 to 70 days of age. Muscle samples from 18 age-matched commercial quail were used as normal controls. Characteristic histological lesions found in the skeletal muscles included sarcoplasmic masses, ringed fibres, internal migration of nuclei and fibre size variation. These lesions, which mainly occurred in the proximal muscles, appeared first in the pectoral region and later in the muscles of the thoracic and pelvic limbs. The most predominant lesion observed at all ages consisted of sarcoplasmic masses. The presence of histological changes did not affect muscle fibre typing by two staining methods, for myosin ATPase at pH 4.5, and by NADH-TR stain. The histological changes were observed in type 2A and less commonly in 2B fibres, but not in type 1. The pectoralis thoracicus muscle, in which lesions were particularly common, showed abnormally large type 2B muscle fibres at 20 days of age. These fibres began to decrease in size at 30 days of age, and at 70 days had become strikingly atrophic, their diameter being only about half that observed at 20 days. The atrophic type 2B muscle fibres were eventually replaced by lipocytes. Chronological staging of the histopathological changes in muscle was impossible since no inter-relationship was observed between the age of the quail, the severity of clinical signs and the extent of muscle lesions. This variability in the severity and age of onset may have been due to the variable expression or incomplete penetrance of the defective gene. Because the disorder is hereditary and progressive in nature, it can be classified as a type of progressive muscular dystrophy.

Maximum shortening velocity and coexistence of myosin heavy chain isoforms in single skinned fast fibres of rat skeletal muscle

Myosin heavy chain composition of a large number (288) of single fibres from slow (soleus), and fast (superficial part of tibialis anterior, and plantaris) muscles of adult (3-5-month-old) Wistar rats was determined. A combination of SDS-PAGE and monoclonal antibodies against myosin heavy chains allowed to identify four myosin heavy chain isoforms (1, 2A, 2X, and 2B) and to detect myosin heavy chain coexistence. Four groups of fibres containing only one myosin heavy chain (1 myosin heavy chain, 2A myosin heavy chain, 2X myosin heavy chain, and 2B myosin heavy chain), and five groups containing more than one myosin heavy chain (1 and 2A myosin heavy chains, 2A and 2X myosin heavy chains, 2X and minor amounts of 2B (2X-2B fibres), 2B and minor amounts of 2X (2B-2X fibres), and 2A, 2X, and 2B myosin heavy chain were identified and their relative percentages were assessed. Coexistence of fast myosin heavy chain isoforms was found to be very frequent (50% of the fibres in plantaris, and 30% in tibialis anterior), whereas coexistence of slow and fast (2A) myosin heavy chain was very rare. Maximum shortening velocity (V0) was determined using the slack-test procedure in a subset of 109 fast fibres from the above population. The values of V0 formed a continuum extending from 2A to 2X to 2X-2B to 2B-2X to 2B fibres. 2A fibres had the lowest value of V0 and 2B fibres the highest. Only the differences between 2A and 2B and 2A and 2B-2X fibres were statistically significant.(ABSTRACT TRUNCATED AT 250 WORDS)

Automated cytometry of fibre size and spatial distribution in the superficial masseter muscle of the rat at three ages.

An image cytometry program was applied to sections of the superficial masseter muscle of female and male 21-, 42- and 105-day-old rats. Lesser diameter and spatial distribution of more than 6000 muscle fibres were automatically measured in digital images from muscle sections stained for myofibrillar ATPase activity. In this muscle, only type 2A, 2B and 2C fibres were observed. At the three ages and in both sexes, 2A fibres were found to have the largest diameter and were the most frequent (> 54%). In the 21-day-old animals, females presented larger diameters than did males; in contrast, in the 105-day-old animals, the three fibre types were larger in males than in females. At all ages and in both sexes, type 2A occupied 32 to 80% more cross-sectional area than type 2B. Most images (98%) showed a random spatial distribution of their fibre types. All fibre types grew in diameter with age. The coefficient of variation of the diameter was age-independent and remained under 23%. The finding of an age-independent variable could have a practical application: an increase of the coefficient of variation (> 25%) can be considered as pathological, even without a perfect age-matched control.

Fibre size, atrophy, and hypertrophy factors in vastus lateralis muscle from 18- to 29-year-old men

In order to study the size of muscle fibres, cross-sections of autopsied vastus lateralis muscle from 8 healthy men, aged 18 to 29 years, who have died suddenly were prepared and analyzed. Data were obtained on cross-sectional area, on lesser diameter, and on atrophy and hypertrophy factors of type 1 and type 2 fibres, subdivided into 2a, 2b, and 2c fibres. The difference in mean fibre size between type 1 and 2 fibres was not significant, whereas the differences between type 1 and 2b, type 1 and 2a, and type 2a and 2b fibres were significant. In the whole material type 2a fibres were the largest and type 2b fibres the smallest. There were considerable differences between post-mortem subjects. Because of these differences and the variability of all fibre types in respect of size in a sample the normal ranges of fibre size were large. The normal ranges and a continuous scale of weights were used to determine the atrophy and hypertrophy factors in each sample, and the upper limits of these factors accepted as being normal. The estimates of the limits of normality of the area, diameter, atrophy and hypertrophy factors of type 1, 2, 2a and 2b fibres reflect the situation in the vastus lateralis muscle of healthy young men. These values might be useful in studying physiological and pathological conditions influencing the size of different fibre types.

DISORDER OF SWALLOWING MUSCLES IN IRON DEFICIENT RABBITS

Plummer-Vinson syndrome is a clinical entity associated with dysphagia, sideropenic anemia and atrophic glossitis. Using rabbits with iron deficiency anemia, the author investigated the cause of dysphagia in this syndrome. Iron deficient animals were produced by bolus feeding without iron and intramuscular injection of an iron-chelating agent. The fibers of swallowing muscles (the thyro-pharyngeal, cricopharyngeal and cervicoesophageal muscles) were classified into three types (Type 1, 2A and 2B fibers) by actomyosin ATPase staining. No significant difference between the muscles of sideropenic rabbits and those of normal rabbits were found in the composition and distribution of their muscle fibers. By NADH-TR staining, however, the disturbance of the intermyofibrillar network and/or a "Moth Eaton" appearance, known to be caused by leakage of mitochondria, were observed in Type 1 fibers of the swallowing muscles of sideropenic rabbits. These morphological changes are similar to those observed in progressive muscular dystrophy. The quantity of iron in the swallowing muscles of sideropenic rabbits was significantly reduced in comparison with that in the sternothyroid and femoral muscles. This finding suggests that a selective decrease in myoglobin occurs in the swallowing muscles of iron deficient animals. From these observations, it might be concluded that dysphagia in iron deficiency anemia is caused by a myogenic disorder.

Fiber type and regional differences in oxidative capacity and glycogen content in the hamster diaphragm.

The purpose of this study was to define variability of the oxidative capacity and glycogen content between different fiber types and regions of the hamster diaphragm. Using histochemical and microphotometric techniques, the oxidative capacity (identified by nicotinamide-adenine dinucleotide tetrazolium reductase reaction end product) and glycogen levels (identified by the periodic acid-Schiff stain test) were examined in three myofibrillar ATPase (M-ATPase) fiber types and four diaphragmatic regions: sternal, anterior costal, thoracic surface of the crural (thor/crur), and abdominal surface of the crural (abd/crur). Most regional differences were found between the crus and the rest of the diaphragm. There were no differences in the oxidative capacity between diaphragmatic regions in the types 1 and 2a fibers, but the type 2b fibers in the thor/crur region had the greatest oxidative capacity and the 2b fiber in the sternal region had the lowest oxidative capacity. There were differences in glycogen content between diaphragmatic regions for all of the three M-ATPase fiber types. Variability in oxidative capacity between fiber types was demonstrated in all regions except the thor/crur region. Variation in glycogen content between fiber types was only demonstrated in the two surfaces of the crus. The type 2b fiber demonstrated the most differences from types 1 and 2a fibers in oxidative capacity and glycogen content in the different diaphragmatic regions, whereas the types 1 and 2a fibers demonstrated few differences from each other in these features across the different diaphragmatic regions.(ABSTRACT TRUNCATED AT 250 WORDS)

Mammalian skeletal muscle fibers distinguished by contents of phosphocreatine, ATP, and Pi.

We tested the proposition that muscle cell types have different contents of phosphocreatine (PCr), ATP, and Pi by 31P NMR spectroscopy and HPLC analyses of adult rat and mouse muscles containing various volume fractions of different fiber types. There was a 2-fold difference in the PCr content between muscles with a high volume fraction of fiber types 1 and 2x versus those with fast-twitch (types 2a and 2b) fiber types. Pi content was low, and PCr and ATP contents were high in muscles with large contents of type 2b and 2a fibers; the reverse was true in muscles with a large volume fraction of type 1 and 2x fibers. There is a large range in the Pi/PCr ratios in normal resting muscles, from less than 0.05 in type 2 to 0.51 in type 1 fibers, depending upon the distribution of their component fiber types. In all muscles, the peak area resulting from the beta phosphate of ATP constituted approximately 13% of the sum of all peak areas observable in the 31P spectrum. Fiber types 2a and 2b were not distinguishable, and the content of type 2x fibers was similar to type 1 fibers. From the profile of these metabolites, we could distinguish only two classes of fibers. For type 2a and 2b fibers, the intracellular concentrations were 8 mM ATP, 39 mM total creatine, 32 mM PCr, 0.8 mM Pi, and 8 microM ADP. For type 1 and 2x fibers, these quantities were 5 mM ATP, 23 mM total creatine, 16 mM PCr, 6 mM Pi, and 11 microM ADP. Thus our results establish an additional criterion upon which to distinguish skeletal muscle cells, one based on the resting content of bioenergetically important metabolites. These results also provide the basis for estimating skeletal muscle fiber-type composition from noninvasive NMR spectroscopic data.