Biomedical subjects

Karla Punkt

Publications and source records attributed to Karla Punkt.

11 recordsLinked to original sources

PubMed2006

Fibre type related changes in the metabolic profile and fibre diameter of human vastus medialis muscle after anterior cruciate ligament rupture.

Vastus medialis muscles of patients with chronic anterior instability of the knee after anterior cruciate ligament rupture were analysed to investigate changes in defined muscle fibres of the diseased leg in comparison to the healthy leg of the same patient. Metabolic and morphological parameters were obtained by cytophotometrical measurements of the activities of succinate dehydrogenase (a marker of oxidative metabolism) and glycerol-3-phosphate dehydrogenase (a marker of glycolytic metabolism) of slow-oxidative (SO), fast-oxidative glycolytic (FOG) and fast-glycolytic (FG) fibre types in serial sections and by measuring the minimal fibre diameters of type I (slow) and type II (fast) fibres. We found decreased glycolytic activity and a shift to more oxidative metabolism in each fibre type suggesting diminished fast force and shift to endurance force development. The latter was interpreted as a sign of active compensation for the knee instability. Significantly decreased minimal fibre diameters to 85.9% in type I fibres, and to 88.7% in type II fibres of the diseased muscle were measured, indicating the fibre atrophy. Our findings suggest that the atrophied muscle fibres of the affected vastus medialis muscle adapt to the altered conditions by changing their metabolic profile. Muscle fibres of different types were found to be affected similarly.

Adolescent↗

PubMed2006

Nitric oxide synthase is up-regulated in muscle fibers in muscular dystrophy.

Nitric oxide (NO) mediates fundamental physiological actions on skeletal muscle. The neuronal NO synthase isoform (NOS1) was reported to be located exclusively in the sarcolemma. Its loss from the sarcolemma was associated with development of Duchenne muscular dystrophy (DMD). However, new studies evidence that all three NOS isoforms-NOS1, NOS2, and NOS3-are co-expressed in the sarcoplasm both in normal and in DMD skeletal muscles. To address this controversy, we assayed NOS expression in DMD myofibers in situ cytophotometrically and found NOS expression in DMD myofibers up-regulated. These results support the hypothesis that NO deficiency with consequent muscle degeneration in DMD results from NO scavenging by superoxides rather than from reduced NOS expression.

Biopsy↗

PubMed2006

Nitric oxide synthase in muscular dystrophies: a re-evaluation.

Duchenne and Becker muscular dystrophies (DMD and BMD) are associated with decreased total nitric oxide (NO). However, mechanisms leading to NO deficiency with consequent muscle-cell degeneration remain unknown. To address this issue, we examined skeletal muscles of DMD and BMD patients for co-expression of NO synthase (NOS) with nitrotyrosine and transcription factor CREB, as well as with enzymes engaged in NO signaling. Employing immunocytochemical labeling, Western blotting and RT-PCR, we found that, in contrast to the most commonly accepted view, neuronal NOS was not restricted to the sarcolemma and that muscles of DMD and BMD patients retained all three NOS isoforms with an up-regulation of the inducible NOS isoform, CREB and nitrotyrosine. We suggest that enhanced nitrotyrosine immunostaining in muscle fibers as well as in the vasculature of DMD and BMD specimens reflects massive oxidative stress, resulting in withdrawal of NO from its regular physiological course via the scavenging actions of superoxides.

3',5'-Cyclic-GMP Phosphodiesterases↗

PubMed2006

Altered fiber distribution and fiber-specific glycolytic and oxidative enzyme activity in skeletal muscle of patients with type 2 diabetes.

OBJECTIVE: We investigated whether alterations of glycolytic and oxidative enzyme capacity in skeletal muscle of patients with type 2 diabetes pertain to specific muscle fibers and are associated with changes in muscle fiber composition. RESEARCH DESIGN AND METHODS: Vastus lateralis muscle was obtained by percutaneous biopsy from 10 patients with type 2 diabetes and 15 age- and BMI-matched healthy volunteers. Using cytophotometry, muscle fiber composition and fiber type-specific glycolytic and oxidative enzyme activities were measured in slow oxidative, fast oxidative glycolytic, and fast glycolytic fibers. RESULTS: In the whole muscle, oxidative activity was decreased in patients with type 2 diabetes. The slow oxidative fiber fraction was reduced by 16%, whereas the fast glycolytic fiber fraction was increased by 49% in skeletal muscle from the diabetic patients. Both oxidative and glycolytic enzyme activities were significantly increased in fast glycolytic and fast oxidative glycolytic fibers of type 2 diabetic patients. However, the fiber-specific ratio of glycolytic enzyme activity relative to oxidative activity was not different between type 2 diabetic patients and the control subjects. The myofibrillic ATP activity was significantly lower in all fiber types of patients with type 2 diabetes and correlates with glucose infusion rate during the steady state of a euglycemic-hyperinsulinemic clamp and maximal aerobic capacity and negatively with HbA(1c) values. CONCLUSIONS: Reduced oxidative enzyme activity in muscle of type 2 diabetic patients is most likely due to a reduction in slow oxidative fibers. Increased glycolytic and oxidative enzyme activities in individual muscle fibers are closely related to measures of long-term glycemic control and whole-body insulin sensitivity and could therefore represent a compensatory mechanism of the muscle in function of the altered glucose metabolism.

Adenosine Triphosphatases↗

PubMed2005

Nitric oxide synthase in human skeletal muscles related to defined fibre types.

Skeletal muscle functions regulated by NO are now firmly established. However, the knowledge about the NO synthase (NOS) expression related to a defined fibre type in human skeletal muscles necessitates further clarification. To address this issue, we examined localization of NOS isoforms I, II and III, in human skeletal muscles employing immunocytochemical labeling with tyramide signal amplification complemented with enzyme histochemistry and Western blotting. The NOS immunoreactivity was related to fibre types of different classification systems: physiological classification into slow and fast, ATPase classification into I, IIA, IIAX, IIX, and physiological-metabolic classification into slow-oxidative (SO), fast-oxidative glycolytic (FOG) and fast-glycolytic (FG). We found a correlation of NOS I-III immunoreactivity to metabolic defined fibre types with strong expression in FOG fibres. This implies that NO as modulator of muscle function is involved in oxidative metabolism in connection with fast force development, which only occurs in FOG fibres. The NOS expression showed no correlation to ATPase fibre subtypes due to the metabolic heterogeneity of ATPase fibre types. Healthy and affected vastus medialis muscles after anterior cruciate ligament rupture revealed similar NOS expression level as shown by Western blotting with, however, different expression patterns related to the fibre types in affected muscles. This suggests an altered modulation of force development in the fibres of diseased muscles.

Adenosine Triphosphatases↗

PubMed2005

Compartmentalization of NO signaling cascade in skeletal muscles.

Skeletal muscle functions regulated by NO are now firmly established. However, the literature on the compartmentalization of NO signaling in myocytes is highly controversial. To address this issue, we examined localization of enzymes engaged in L-arginine-NO-cGMP signaling in the rat quadriceps muscle. Employing immunocytochemical labeling complemented with tyramide signal amplification and electron microscopy, we found NO synthase expressed not only in the sarcolemma, but also along contractile fibers, in the sarcoplasmic reticulum and mitochondria. The expression pattern of NO synthase in myocytes showed striking parallels with the enzymes engaged in L-arginine-NO-cGMP signaling (arginase, phosphodiesterase, and soluble guanylyl cyclase). Our findings are indicative of an autocrine fashion of NO signaling in skeletal muscles at both cellular and subcellular levels, and challenge the notion that the NO generation is restricted to the sarcolemma.

Animals↗

PubMed2004

Differentiation of rat skeletal muscle fibres during development and ageing.

The purpose of the present study was to determine at which point in the period from embryonic day 21 up to postnatal day (PD) 75, the different fibre types and subtypes are detectable in rat extensor digitorum longus, soleus and gastrocnemius muscles using immunohistochemical, enzyme histochemical and cytophotometrical methods. Moreover, fibre type-specific changes in metabolic profile and changes in fibre type population during postnatal development were analysed. Before birth, no clear differentiation of fibre types was found. At PD 1, slow and fast fibres were typed by antibodies against neonatal, slow and fast myosin heavy chains (MHCs). At PD 8, the different ATPase activities of slow and fast MHCs after alkaline preincubation were detected histochemically. At PD 21, differences in acid stability of ATPase activity of fast MHC isoforms revealed the fast subtypes IIA and IIB (including IIX). At this age, also differences in metabolic properties (oxidative and glycolytic enzyme activities) of fibres were detected for the first time by cytophotometry classifying the fibres into SO, FOG I, FOG II and FG. Before the age of 21 days, the fast fibres were metabolically undifferentiated. During further development and ageing, the population of FG fibres with high glycolytic activity increased at the expense of FOG fibres suggesting FOG to FG transformation. Cytophotometrical measurements revealed that the muscle fibres developed their highest contractile, oxidative and glycolytic activity at PD 21, the time of weaning. In this way, muscle fibres may be prepared for the higher demands for posture and mobility after leaving the nest.

Aging↗

PubMed2002

Nitric oxide synthase II in rat skeletal muscles.

Constitutive expression of nitric oxide synthase (NOS) II was found in rat hindlimb muscles by immunohistochemistry and western blotting during development from embryonic day 21 to the adult stage of 75 days. The immunohistochemical NOS II expression pattern was related to the physiological metabolic fibre types SO (slow-oxidative), FOG I, II (fast-oxidative glycolytic; I more glycolytic, II more oxidative) and FG (fast-glycolytic) and to the myosin-based fibre types I and IIA, IIB (IIX not separated) identified in serial sections by enzyme histochemistry and immunohistochemistry. In adult muscles only the small population of FOG II fibres, which is a part of both IIA and IIB fibre population, showed NOS II immunoreactivity. This is the reason that only weak NOS II expression in adult hindlimb muscles has been detected by western blotting. Hindlimb muscles of embryonic, neonatal and young rats of 8 days expressed more NOS II as compared with adult rat hindlimb muscles. This can be explained by the findings that before the age of 21 days fast fibres were metabolically undifferentiated, all of them were NOS II positive and contribute to the NOS II expression of the muscle. In muscles of diabetic rats the NOS II expression was elevated indicating an inhibition of glucose uptake into the muscle fibres of diabetic muscles. Our findings suggest that the NOS II may be designated both as constitutive and inducible.

Adenosine Triphosphatases↗

PubMed2002

Fibre types in skeletal muscles.

This book is a concise summary of the present knowledge about skeletal muscle fibres. The fibre types were characterized from different points of view. The difficulties and possibilities of classifying muscle fibres in distinct non-overlapping types were shown. A main emphasis is put on metabolic fibre typing by cytophotometry as well as the adaptability of a given fibre type to altered physiological and pathological conditions. Extensive analyses of several rat hindlimb muscles revealed regional differences of fibre properties within the muscles and showed the influence of ageing, myopathy, hypoxia, diabetes and Ginkgo biloba-treatment on the different fibre types. The plasticity of muscle fibres was demonstrated by the flexibility of fibre metabolism as an adaptive mechanism.

Animals↗

PubMed2002

Myopathy-dependent changes in activity of ATPase, SDH and GPDH and NOS expression in the different fibre types of hamster muscles.

Proximal (vastus lateralis) and distal (gastrocnemius) muscles of 100-day-old normal and myopathic BIO TO-2 hamsters were analysed to study the effects of myopathy on the different muscle fibre types: SO (slow oxidative), FOG (fast oxidative glycolytic) and FG (fast glycolytic). Cytophotometric measurements of enzyme activities (myofibrillic adenosine triphosphatase, succinate dehydrogenase and glycerol-3-phosphate dehydrogenase), Western blot analysis of nitric oxide synthase (NOS) I, II, III isoforms and NOS II immunohistochemistry were performed. The following alterations were found in myopathic muscle fibres: all fibre types of both proximal and distal myopathic muscles showed decreased myofibrillic adenosine triphosphatase activity indicating depressed contractility. This was associated with depressed oxidative activity of the muscle fibres. A shift to more glycolytic metabolism was observed, mainly in FG fibres of proximal muscle. We found an increased NOS II expression in both myopathic muscle types investigated. It means that increased NO production inhibits force generation in myopathic muscle. NOS II immunoreactivity was found mainly in the cytoplasm of FG fibres. NOS I and NOS III expression was not significantly effected by this form of myopathy. Our findings demonstrate that muscle fibres of proximal and distal skeletal muscles of 100-day-old cardiomyopathic BIO TO-2 hamsters are altered with respect to contractility, metabolism and NOS II expression. FG fibres of the proximal muscle were effected most strongly.

Adenosine Triphosphatases↗

PubMed2002

Cytophotometrical and immunohistochemical analysis of soft palate muscles of children with isolated cleft palate and combined cleft lip and palate.

Palatal muscle biopsies from the cleft margin of children were subjected to cytophotometrical and immunohistochemical analysis. Muscle fiber types were classified according to the enzyme activity of myofibrillic adenosine triphosphatase, glycerol-3-phosphate-dehydrogenase and succinate dehydrogenase assessed cytophotometrically. Fiber type-related immunoreactivity of nitric oxide synthase (NOS) isoforms I, II, III, as a physiological modulator of skeletal muscle function, was related to the oxidative and glycolytic activity of the muscle fibers. Fast oxidative glycolytic fibers with high oxidative activity showed strong NOS I immunoreactivity, whereas fast glycolytic fibers with high glycolytic activity were stronger immunolabelled for NOS III. NOS II expression was similar in all fiber types. No differences in NOS immunoreactivity were found between the two investigated forms of deformity. Additionally to the usual skeletal muscle fiber types, a slow tonic fiber type was for the first time identified in cleft palate muscles. Comparison of two forms of cleft palate, isolated cleft palate and combined cleft lip and palate has shown decreased enzyme activities in muscle fibers of palatal muscles from combined cleft lip and palate. Fast oxidative glycolytic fibers were mainly effected. Cytophotometrical and immunohistochemical analysis indicated a depressed performance of the cleft palatal muscles from combined cleft lip and palate as a stronger deformity compared with isolated cleft palate.

Adenosine Triphosphatases↗