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Characterization of a mammalian smooth muscle myosin heavy chain cDNA clone and its expression in various smooth muscle types.

A cDNA clone, SMHC-29, encoding the light meromyosin of smooth muscle myosin heavy chain (MHC), was isolated from a rabbit uterus cDNA library constructed in phage lambda gt11. This smooth muscle MHC cDNA demonstrates significant nucleotide and amino acid sequence homologies with known sarcomeric MHC genes from rabbit, rat skeletal, and nematode body wall myosin, and even with nonmuscle MHC gene from a slime mold (Dictyostelium discoideum), suggesting that smooth muscle, striated muscle, and nonmuscle MHC genes diverged from a common ancestor. The deduced amino acid sequences of the smooth muscle light meromyosin show very similar periodic distributions of hydrophobic and charged residues as found for the light meromyosin of striated muscle MHCs together with a high potential for alpha-helical formation, indicating an alpha-helical coiled-coil structure for the smooth muscle light meromyosin sequences. Furthermore, S1 nuclease mapping has revealed that this smooth muscle MHC gene for SMHC-29 is specifically expressed in smooth muscles of vascular and nonvascular types but not in the striated muscles or nonmuscle cells.

Amino Acid Sequence↗

Skeletal muscle Na,K-ATPase alpha and beta subunit protein levels respond to hypokalemic challenge with isoform and muscle type specificity.

During potassium deprivation, skeletal muscle loses K+ to buffer the fall in extracellular K+. Decreased active K+ uptake via the sodium pump, Na,K-ATPase, contributes to the adjustment. Skeletal muscle expresses alpha1, alpha2, beta1, and beta2 isoforms of the Na, K-ATPase alphabeta heterodimer. This study was directed at testing the hypothesis that K+ loss from muscle during K+ deprivation is a function of decreased expression of specific isoforms expressed in a muscle type-specific pattern. Isoform abundance was measured in soleus, red and white gastrocnemius, extensor digitorum longus, and diaphragm by immunoblot. alpha2 expression was uniform across control muscles, whereas alpha1 and beta1 were twice as high in oxidative (soleus and diaphragm) as in fast glycolytic (white gastrocnemius) muscles, and beta2 expression was reciprocal: highest in white gastrocnemius and barely detectable in soleus and diaphragm. Following 10 days of potassium deprivation plasma K+ fell from 4.0 to 2.3 mM, and there were distinct responses in glycolytic versus oxidative muscles. In glycolytic white gastrocnemius alpha2 and beta2 fell 94 and 70%, respectively; in mixed red gastrocnemius and extensor digitorum longus both fell 60%, and beta1 fell 25%. In oxidative soleus and diaphragm alpha2 fell 55 and 30%, respectively, with only minor changes in beta1. Although decreases in alpha2 and beta2 expression are much greater in glycolytic than oxidative muscles during K+ deprivation, both types of muscle lose tissue K+ to the same extent, a 20% decrease, suggesting that multiple mechanisms are in place to regulate the release of skeletal muscle cell K+.

Animals↗

Cell cycle-mediated regulation of smooth muscle alpha-actin gene transcription in fibroblasts and vascular smooth muscle cells involves multiple adenovirus E1A-interacting cofactors.

Expression of smooth muscle alpha-actin in growth factor-induced myofibroblasts and in differentiated vascular smooth muscle cells is transcriptionally controlled by multiple positive or negative trans-acting factors interacting with distinct cis-elements in the 5'-flanking region of the gene. Because none of the transcriptional regulators reported to date is smooth muscle cell- or myofibroblast-specific per se, the dynamic interplay among many factors interacting at specific sites along the promoter appears to be a signature feature of smooth muscle alpha-actin gene regulation in these cell types. Herein, the ability of the adenovirus E1A 12 S protein to bind and functionally inactivate specific cell regulatory factors has been exploited to identify several previously unknown coactivators of the mouse smooth muscle alpha-actin promoter in rodent fibroblasts and vascular smooth muscle cells. In transient cotransfection assays, ectopic expression of wild type E1A suppressed promoter activity in a dose- and cis-element-dependent manner. In asynchronous cells, N-terminal E1A mutants defective in CREB-binding protein (CBP) and p300 binding capacity exhibited markedly reduced inhibitory activity toward a smooth muscle alpha-actin promoter driven by a composite TEF-1-, SRF-, and Sp1/3-regulated enhancer. In synchronized cells, however, a more complex mutant E1A inhibitory pattern indicated that collaboration between CBP/p300 and the retinoblastoma family of pocket proteins was required to produce a fully functional enhancer. Cotransfection experiments conducted with Rb(-/-) fibroblasts demonstrated the necessity of pRB in augmenting smooth muscle alpha-actin enhancer/promoter activity. Physical interaction studies with the use of purified wild type and mutant E1A proteins confirmed that CBP, p300, and pRB were targets of E1A binding in nuclear extracts of vascular smooth muscle cells and/or fibroblasts. Collectively, these results suggest that a repertoire of E1A-interacting proteins, namely CBP/p300 and pRB, serve to integrate the activities of multiple trans-acting factors to control smooth muscle alpha-actin gene transcription in a cell type- and cell cycle-dependent manner.

Actins↗

Three-dimensional reconstruction of the rat triceps surae muscle and finite element mesh generation of the gastrocnemius medialis muscle.

In order to simulate blood flow in skeletal muscle, our group has developed a finite element description of perfused skeletal muscle. This model requires input parameters concerning the vascular system, muscle contraction and the geometry of a muscle, including its aponeuroses. The objective of the present paper is to create a geometrical reconstruction of the rat gastrocnemius medialis muscle that can be incorporated in the finite element model. Since this muscle is connected to the plantaris and the gastrocnemius lateralis muscle, a detailed computer graphical reconstruction of the triceps surae muscle, based on histological cross-sections, has been accomplished first. Using this reconstruction, relevant sections were selected to create the finite element mesh of the gastrocnemius medialis muscle. Special attention was payed to the location of the aponeuroses. The mesh can be used in finite element simulations of perfused skeletal muscle.

Animals↗

Expression of muscle-gene-specific isozymes of phosphorylase and creatine kinase in innervated cultured human muscle.

Isozymes of creatine kinase and glycogen phosphorylase are excellent markers of skeletal muscle maturation. In adult innervated muscle only the muscle-gene-specific isozymes are present, whereas aneurally cultured human muscle has predominantly the fetal pattern of isozymes. We have studied the isozyme pattern of human muscle cultured in monolayer and innervated by rat embryo spinal cord explants for 20-42 d. In this culture system, large groups of innervated muscle fibers close to the ventral part of the spinal cord explant continuously contracted. The contractions were reversibly blocked by 1 mM d-tubocurarine. In those innervated fibers, the total activity and the muscle-gene-specific isozymes of both enzymes increased significantly. The amount of muscle-gene-specific isozymes directly correlated with the duration of innervation. Control noninnervated muscle fibers from the same dishes as the innervated fibers remained biochemically immature. This study demonstrated that de novo innervation of human muscle cultured in monolayer exerts a time-related maturational influence that is not mediated by a diffusable neural factor.

Animals↗

A comparative study on the fine structure of the basalar muscle of the wing and the tibial extensor muscle of the leg of the lepidopteran Achalarus lyciades.

Basalar and tibial extensor muscle fibers of Achalarus lyciades were examined with light and electron microscopes. Basalar muscle fibers are 100-150 micro in diameter. T-system membranes and sarcoplasmic reticulum make triadic contacts midway between Z lines and the middle of each sarcomere. The sarcoplasmic reticulum is characterized by a transverse element situated among myofilaments halfway between Z lines in every sarcomere. The morphology of Z lines, hexagonal packing of thin and thick myofilaments, and thin/thick myofilament ratios are similar to those of fast-acting insect muscles. Tibial extensor muscle fibers are 50-100 micro in diameter. Except for a lack of the transverse element, the T system and sarcoplasmic reticulum are similar to those of basalar muscle. Wavy Z lines, lack of a hexagonal packing of myofilaments, and larger thin/thick myofilament ratios are similar to those of other postural muscles of insects. The morphology of basalar and tibial extensor muscle is compared to that of other insect muscle with known functions, and reference is made to the possible contribution of the transverse element of sarcoplasmic reticulum in basalar flight muscle to speed and synchrony in this muscle.

Animals↗

Muscle fibres and cultured muscle cells express the B7.1/2-related inducible co-stimulatory molecule, ICOSL: implications for the pathogenesis of inflammatory myopathies.

Inducible co-stimulator ligand (ICOSL), a member of the B7 family of co-stimulatory molecules related to B7.1/2, regulates CD4 as well as CD8 T-cell responses via interaction with its receptor ICOS on activated T cells. Here we examined the expression and the functional relevance of ICOSL in human muscle cells in vivo and in vitro. We investigated 25 muscle biopsy specimens from patients with polymyositis, dermatomyositis, inclusion body myositis, Duchenne muscular dystrophy and non-myopathic controls for ICOSL expression by immunohistochemistry. Normal muscle fibres constitutively express low levels of ICOSL. However, ICOSL expression is markedly increased in muscle fibres in inflammatory myopathies. Cell surface staining was most prominent in the contact areas between muscle fibres and inflammatory cells, which in turn show expression of ICOS as a marker of T-cell activation. Muscle endothelial cells show constitutive expression of ICOSL under normal and pathological conditions. We also detected mRNA and cell surface protein expression of ICOSL on myoblasts cultured from control subjects and patients as well as in TE671 muscle rhabdomyosarcoma cells. ICOSL expression was upregulated by tumour necrosis factor-alpha (TNF-alpha), whereas interferon-gamma (IFN-gamma) had no such effect. Co-culture experiments of major histocompatibility complex (MHC) class II-positive myoblasts with CD4 T cells together with superantigen demonstrated that the expression of muscle-related ICOSL has functional consequences: the production of Th1 (IFN-gamma) and Th2 cytokines [interleukin (IL)-4 and IL-10] by CD4 T cells was markedly reduced in the presence of a neutralizing anti-ICOSL monoclonal antibody (mAb HIL-131), thus showing the importance of ICOSL co-stimulation for T-cell activation. Taken together, our results demonstrate that human muscle cells express ICOSL, a functional co-stimulatory molecule distinct from B7.1 and B7.2. ICOSL-ICOS interactions may play an important role in inflammatory myopathies, providing further evidence for the antigen-presenting capacity of muscle cells.

Adult↗

Muscle-specific alternative splicing of myotubularin-related 1 gene is impaired in DM1 muscle cells.

The myotubularin-related 1 (MTMR1) gene belongs to a highly conserved family of eucaryotic phosphatases, with at least 11 members in humans. The founder member of this gene family, MTM1, is mutated in X-linked myotubular myopathy, a severe congenital disorder that affects skeletal muscle, and codes for myotubularin, a specific phosphatidylinositol 3-phosphate [PI(3)P] phosphatase. MTM1 and MTMR1 are adjacent on the X chromosome, and the corresponding proteins share 59% sequence identity. In the present study, we investigated the putative role of MTMR1 in myogenesis by analysing its expression pattern in muscle cells during differentiation and in skeletal muscle throughout development. We have identified three novel coding exons in the MTMR1 intron 2 that are conserved between mouse and human, are alternatively spliced, and give rise to six mRNA isoforms. One of the transcripts is muscle-specific and is induced during myogenesis both in vitro and in vivo, and represents the major isoform in adult skeletal muscle. We show that the two main MTMR1 protein muscular isoforms, like myotubularin, efficiently dephosphorylate PI(3)P in vitro. We have also analysed whether MTMR1 alternative splicing is affected in skeletal muscle cells derived from patients with congenital myotonic dystrophy (cDM1), in which mRNA splicing disturbances of specific genes are thought to constitute an important pathogenic mechanism. We found a striking reduction in the level of the muscle-specific isoform and the appearance of an abnormal MTMR1 transcript in differentiated cDM1 muscle cells in culture and in skeletal muscle from cDM1 patients. Our results suggest that MTMR1 plays a role in muscle formation and represents a novel target for abnormal mRNA splicing in myotonic dystrophy.

Alternative Splicing↗

Tetrahymena actin: copolymerization with skeletal muscle actin and interactions with muscle actin-binding proteins.

We have previously shown that actin from Tetrahymena pyriformis has a very divergent primary structure (Hirono, M., Endoh, H., Okada, N., Numata, O., & Watanabe, Y. (1987) J. Mol. Biol. 194, 181-192) and that though it shares essential properties with skeletal muscle actin, it does not interact at all with phalloidin or DNase I (Hirono, M., Kumagai, Y., Numata, O., & Watanabe, Y. (1989) Proc. Natl. Acad. Sci. U.S. 86, 75-79). In this study, we investigated the copolymerization of this actin with skeletal muscle actin by direct observation of the heteropolymers formed from the two actins by means of electron microscopy. We also examined the binding of actin-binding proteins from skeletal muscle or smooth muscle to Tetrahymena actin by means of a cosedimentation assay. The results show that (i) Tetrahymena actin copolymerizes with skeletal muscle actin and that (ii) muscle myosin subfragment 1 binds to it in the absence of ATP, like skeletal muscle actin. However, it was also shown that (iii) muscle alpha-actinin hardly binds to Tetrahymena actin and that (iv) muscle tropomyosin does not bind to it at all. The results show that Tetrahymena actin has both properties similar and dissimilar to those of skeletal muscle actin.

Actinin↗

Influence of selection for breast muscle mass on myosin isoform composition and metabolism of deep pectoralis muscles of male and female turkeys.

Advances in genetic selection and nutrition have resulted in rapid growth rates and increased muscle mass, predisposing turkeys to muscle disorders such as deep pectoral myopathies and increasing the incidence of pale, soft, and exudative muscle. The objective of this study was to determine if selection for breast muscle mass created an increase in anaerobic capacity of the deep pectoralis muscle. A total of 67, 18-wk-old, male and female turkeys from two male (tom) lines and one female (hen) line were used. Each bird was anesthetized and one deep pectoralis muscle was electrically stimulated via the pectoral nerve. Muscle pH was recorded every 30 s for 4 min of stimulation and every 1 min for a 10-min recovery period. Non-stimulated muscles, contralateral to the stimulated side, were assayed for lactate dehydrogenase (LDH) and glyceraldehyde phosphate dehydrogenase (GAPDH). Myosin isoforms were resolved with SDS-PAGE. Line or gender had no effect on rate of pH decline during or after stimulation. Declines in pH during stimulation were greater than during the recovery period (0.06 vs. 0.02 U/min). The lightweight male line (LM) had the greatest breast muscle mass as a percentage of body weight (P < 0.05) and the greatest LDH [293 mmol nicotinamide adenine dinucleotide (NADH) min(-1)microg(-1); P < 0.0001] and GAPDH (0.4452 mmol NADH min(-1)microg(-1); P < 0.05) activities. Hens had greater percentages breast weight than males (P < 0.05) and a tendency for increased enzyme activities. The LM line had the largest ratio (2.33:1) (P < 0.05) of adult-to-neonatal myosin. Genetic selection for breast muscle mass resulted in an increased ratio of adult-to-neonatal myosin and increased anaerobic capacity. This effect on myosin isoform composition and anaerobic capacity supports handling modifications that are line specific to minimize meat quality defects.

Anaerobic Threshold↗

Overexpression of muscle uncoupling protein 2 content in human obesity associates with reduced skeletal muscle lipid utilization.

Uncoupling proteins (UCP) may influence thermogenesis. Since skeletal muscle plays an important role in energy homeostasis and substrate oxidation, this study was undertaken to test the hypotheses that skeletal muscle UCP2 content is altered in obesity and could be linked to basal energy expenditure, insulin sensitivity, or substrate oxidation within skeletal muscle under postabsorptive (fasting) conditions. To examine these possibilities, limb basal energy expenditure and respiratory quotient (bRQ) were measured in 18 obese nondiabetic (Ob) and lean individuals (L). Total body fat (%) ranged from 11% to 46%. In addition, insulin-stimulated rates of glucose disposal (Rd) were measured under euglycemic hyperinsulinemic conditions. Biopsy of vastus lateralis muscle was used to measure cytochrome c oxidase (COX) enzyme activity and UCP2 content. Whereas low muscle COX activity was found in the Ob compared to L (6.9+/-1.6 vs. 9.6+/-1.2 U/g; P<0.001), skeletal muscle UCP2 content in Ob was significantly higher than in L (48+/-9 vs. 33+/-12 arbitrary units/g; P<0.05). Moreover, UCP2 content was positively correlated with percent of total body fat (r=0.57; P<0. 05) and bRQ (r=0.59; P<0.01), but not with visceral fat (r=0.17; P=0. 49), basal energy expenditure (r=0.07; P=0.79) or Rd (r=-0.23; P=0. 34). In summary, these results indicate that if development of obesity in humans is mediated by defective expression of UCP2 within skeletal muscle, then this effect is not observed in people with established obesity. The present study also suggests that skeletal muscle UCP2 content is not related to basal energy expenditure or insulin sensitivity in humans. However, the increased content of UCP2 within skeletal muscle in obesity appears to coincide with a reduced postabsorptive lipid utilization by muscle.

Adult↗

An experimental study of skin flap associated with muscle: is muscle nourishment possible through the musculocutaneous perforators?

An experimental study was planned to examine whether the blood supply of muscle would be maintained by reverse flow from the cutaneous arteriolar microcirculatory system via the musculocutaneous perforators. A flap model containing both muscle and skin based on the inferior superficial epigastric vessels was designed with the blood supplied directly from the cutaneous arteriolar microcirculatory system. A total of 154 male Wistar Albino rats were divided into three groups. Group I included the standard vertical rectus abdominis musculocutaneous flap based on the superior deep epigastric vessels (N = 48). Group II included the acute cutaneous muscle flap (N = 53). Group III contained the delayed cutaneous muscle flap (N = 53). Skin flap survival area, muscle scintigraphy with technetium-99m-methoxy-isobutyl-isonitrile, microangiography, and histopathological examination of the flaps were conducted. The mean percentage of surviving skin paddle area was 96.4 +/- 5.2%, 84.9 +/- 21.6%, and 91.0 +/- 16.8% in groups I, II, and III respectively. There was no significant difference between groups. Microangiography revealed the blood flow from skin to muscle through the musculocutaneous perforators. The radioisotope uptake of the muscle flap was expressed as A percentage of the intact contralateral muscle. Mean uptake in group I was 90.1 +/- 4.9% immediately after flap elevation, 62.5 +/- 13.5% on day 2, and 88.3 +/- 12.0% on day 7. These values were 53.7 +/- 7.1%, 63.6 +/- 14.1%, and 89.2 +/- 18.1% in group II, and 64 +/- 7.8%, 75.5 +/- 9.8%, and 92.8 +/- 40.1% in group III. Radioisotope uptake in group I was significantly higher than groups II and III immediately after flap elevation (p < 0.05, analysis of variance), whereas there was no significant difference on days 2 and 7. Histopathological examination revealed surviving muscle tissue without marked atrophy. There was no marked difference between groups histopathologically. These results indicate that muscle tissue may survive by reverse flow through the musculocutaneous perforators when elevated with an axial skin flap.

Animals↗

The role of the gastrocnemius muscle flap in limb-sparing surgery for bone sarcomas of the distal femur: a proposed classification of muscle transfers.

Limb-sparing surgery for bone and soft-tissue sarcomas involves three phases: (1) resection of the tumor with free margins, (2) reconstruction of the bone and joint defect, and (3) reconstruction of the soft tissues. This presentation focuses on the third phase. Between January of 1988 and January of 1994 we performed 33 distal femoral resections, 32 for malignant and 1 for benign tumors. There were 19 men and 14 women aged from 6 to 78 years (mean age 28 years). Twenty-seven patients had gastrocnemius muscle flap transfers, 24 having "primary" transfers and 3 having "secondary" transfers. The lateral gastrocnemius muscle was used in 18 patients, the medial in 8 patients, and both in 1 patient. We propose a classification of the transfers based on the size of the soft-tissue defect above the prosthesis needing coverage and the length of the neurovascular bundle of the muscle. Twenty-six of the 27 muscles survived; one patient had necrosis of the skin and muscle. Two patients had persistent sinuses at the scar that were managed successfully (one of them was before a secondary muscle transfer). Six patients did not have gastrocnemius muscle flap transfers. Two of them had persistent sinuses for years, and one patient had titanium "synovitis" and needed repeated operations including removal of the prosthesis and revision. The particular vascularization of the gastrocnemius muscle (one pedicle at the level of the knee joint) situated close to its origin, the size of the muscle belly, and the fact that it is situated in the dissection field and its transfer does not affect the function of the spared limb make it particularly suitable for the coverage of wide areas of skin and muscle loss at the knee region. In the classification that we propose, type I is reserved for coverage of small areas, while types II and III are used for coverage of larger areas. Flap transfer should be performed primarily at the time of the resection in order to avoid complications of wound healing and to reduce delays in chemotherapy protocols.

Adult↗

Detailed neurovascular anatomy of the serratus anterior muscle: implications for a functional muscle flap with multiple independent force vectors.

A functional muscle free flap with multiple muscle segments that could be oriented independently to produce different force vectors would be beneficial in facial reanimation and upper extremity reconstruction. The serratus anterior muscle has this potential because two or more individual muscle slips can be transferred on a single vascular pedicle. Although serratus anterior muscular anatomy has been studied previously, little attention has been given to the intramuscular anatomy. Muscle slips 5 through 9 (and 10, if present) in 50 specimens from 27 cadavers were studied following intraarterial latex injection. Eight specimens were injected with a radiopaque material (latex/diatrizoate/lead mixture) for x-ray delineation of the intramuscular vascular pattern. Slips 5 through 9 are consistently supplied by a single dominant branch of the thoracodorsal artery and innervated by the long thoracic nerve. Dissection revealed that the long thoracic nerve and its branches invariably follow the artery and divide proximal to the corresponding arterial division. There is a consistent vascular pattern to each muscle slip, in which the serratus artery gives rise to common slip arteries, each of which supplies adjacent muscle slips. The mean length of a muscle slip from its origin on the rib periosteum to the division of the common slip artery is 9.6 cm. These findings imply that the slips may be separated to the level of these common slip arteries, with up to five slips transferred on a single neurovascular pedicle and each slip oriented independently to provide multiple muscle force vectors. With these possibilities, the reconstructive surgeon may be able to restore more natural facial animation and better intrinsic muscle function in the upper extremity.

Adolescent↗

Comparison of myosins from the masseter muscle of adult rat, mouse and guinea-pig. Persistence of neonatal-type isoforms in the murine muscle.

Adult rat, mouse, and guinea-pig masseter muscles display distinct myosin electrophoretic patterns. The rat muscle contains four main forms which by reference to the myosins of the IIB tensor fasciae latae, of the IIA mylohyoid, and of the red and white portions of the sternomastoid muscles, correspond respectively to the intermediate-type and to the three fast-type isoforms. The mouse masseter muscle contains only three main myosins, the intermediate-type and two fast-type isoforms. The guinea-pig muscle also displays only three bands, whose assignment is, however, less straightforward than in the murine species; their electrophoretic mobilities are not strictly the same as those of their homologous forms in rat and mouse. Comparison with the myosins of the tensor fasciae latae and of the sternomastoid muscles of guinea-pig allows their identification as intermediate and fast-type myosins. In addition to these typical adult-type forms, adult murine masseter muscles are observed to contain between zero and 30% of neonatal-type myosins. The comparison of the developmental transitions of myosins in the rat masseter with those in the skeletal muscles of the same animal indicates a delay in the appearance of the adult as well as in the disappearance of the neonatal-type myosins in the masseter muscle. Both the variability in myosin types with the animal species and the atypical presence of neonatal forms in the murine adults suggest that myosin expression in the masseter muscle is subjected to unusual regulations.

Aging↗

Muscle metabolic changes associated with long-term inhalation anaesthesia in the horse analysed by muscle biopsy and microdialysis techniques.

During anaesthesia in the horse, muscle blood flow has been found to be reduced, possibly leading to hypoxia or ischaemia in the muscle. The aim of this study was to use the muscle biopsy and microdialysis techniques to determine whether long-term inhalation anaesthesia in laterally recumbent horses induces metabolic changes in gluteal muscle indicative of anaerobic metabolism. Muscle biopsies and plasma samples were taken from seven horses at the start and end of halothane anaesthesia. In six isoflurane-anaesthetised horses, given three pharmacological provocations (dobutamine, detomidine, acepromazine), repeated blood samples and microdialysis was performed during anaesthesia and muscle biopsies were taken before and at the end of anaesthesia. Adenosine triphosphate (ATP), adenosine diphosphate, adenosine monophosphate, inosine monophosphate (IMP) creatine phosphate and lactate concentrations did not differ between dependent and non-dependent muscles at either sampling time. Creatine phosphate decreased in both the halothane (-38%) and isoflurane (-28%) group. In the halothane group, ATP was decreased (-15%) at the end of anaesthesia, while IMP was increased (+32%). Lactate in muscle and plasma increased in both groups. Lactate in dialysate increased after induction and remained elevated above plasma concentrations. These results show that long-term inhalation anaesthesia in horses is associated with an anaerobic metabolic response within the muscle and that microdialysis can be used to detect metabolic changes within the muscle during equine anaesthesia.

Anesthesia, Inhalation↗

Human myocardial and skeletal muscle enzyme activities: creatine kinase and its isozyme MB as related to citrate synthase and muscle fibre types.

Activities of myocardial and skeletal muscle total creatine kinase (CK) and its isozyme MB were related to the oxidative capacity [measured as the citrate synthase (CS) activity] and to the contractile characteristics (estimated as the percentage of type I muscle fibres). Skeletal muscle biopsies were obtained both from physically trained and untrained men and myocardial biopsies from patients subjected to open-heart surgery performed because of mitral or aortic valve disease. Enzyme activities were determined on freeze-dried muscle specimens. The CK-MB activity was about twice as high in trained skeletal muscle as in untrained ones reaching the myocardial level. The total CK activity was about three times higher in skeletal muscle than in myocardium; the myocardium, however, had CS activity 3-4 times larger than that of skeletal muscle. A close correlation was demonstrated between activities of CK-MB on one hand and CS (r = 0.76) or percentage type I fibres (r = 0.83) on the other hand suggesting a connection between CK-MB activity and the oxidative capacity of the cell. This was in contrast to total CK where different regressions were obtained when comparing the myocardium and the skeletal muscle of trained or untrained men. In conclusion, CK-MB activity in trained skeletal muscle in athletes were similar to that in myocardium. CK-MB was related to the oxidative capacity and formation of cellular energy in skeletal and heart muscle.

Adult↗

Capillary supply of the muscle fibre population in hindlimb muscles of the cat.

Comparative analyses of the fibre content (FG, FOG, and SO fibres) and the capillary density (the number of capillaries surrounding individual fibres and the capillary/fibre ratio) were performed in hind limb muscles of the cat. Cross-sections from the tenuissimus, the biceps femoris, the lateral head (LG) and the medial head (MG) of the gastrocnemius and the soleus were cut in a cryostat. The sections were stained histochemically for the NADH2-diaphorase and alkaline (pH 9.4) actomyosin ATPase activity, which enables differentiation of different types of fibres. The endothelium of the capillaries was identified via staining for unspecific alkaline ATPase activity. The number of capillaries surrounding each individual muscle fibre had a positive correlation, first to the oxidative capacity and secondly to the average diameter of the fibres. The thin tenuissimus muscle did not differ in this respect from the thicker muscles. The highest proportion of SO fibres was found in the soleus and the MG muscles. FG fibres of two different types were dominating the fibre mass in the biceps femoris and the LG muscles, while the tenuissimus contained more FOG fibres than these muscles. In general the FG fibres had a larger diameter than the FOG and the SO fibres. The soleus and the MG muscles contained larger fibres than the other examined muscles. FG fibres were surrounded by fewer capillaries than FOG and SO fibres. The soleus and the MG muscles, with a higher percentage of SO fibres and also larger fibres, had the largest number of capillaries around the fibres and the highest capillary/fibre ratio.

Animals↗