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Biomedical subjects

R Billeter

Publications and source records attributed to R Billeter.

At least 37 records · Page 2Linked to original sources
PubMed1996

Design of the oxygen and substrate pathways. VI. structural basis of intracellular substrate supply to mitochondria in muscle cells.

This paper quantifies the structures involved in the transport and oxidation of carbohydrates and fatty acids within the muscle cell. The structural capacity is measured on whole-body random samples of the musculature of dogs and pygmy goats and compared with maximal rates of oxygen consumption and substrate oxidation. Comparing dogs and goats of the same body size provided a 1.55-fold difference in the maximal rate of oxidation when related to muscle mass. As in previous studies, we found that the volume of mitochondria was approximately proportional to aerobic capacity. The maximal glucose flux from intracellular stores to mitochondria is 1.6 times greater in the dog than in the goat; we find that the amount of glycogen stored in the muscle cells is 4.2 times as great in the dog, but part of the intracellular glycogen pool is used for anaerobic rather than for oxidative metabolism. The maximal fatty acid flux from intracellular stores to mitochondria is 1.5 times larger in the dog, and the amount of lipid stored is 2.3 times as great in the dog. Every lipid droplet is in direct contact with the outer membrane of a mitochondrion and the contact surface area is 3.6 times greater in the dog than in the goat. Additional measurements are needed to investigate the role of structural limitation at this step. The amount of substrates stored intracellularly in the muscle cells of the dog is about twice as much as would match the differences in the maximal rates of utilization. This allows the endurance-specialized dogs to run for longer periods at higher rates of oxidation.

Animals↗
PubMed1995

Ultrastructural modification of human skeletal muscle tissue with 6-month moderate-intensity exercise training.

The study was aimed at investigating if endurance training of moderate intensity and longer duration, intended to promote health rather than performance, evokes ultrastructural changes in skeletal muscle tissue comparable to those observed after high-intensity protocols. Twenty healthy, middle-aged men enrolled in a 6-month, home-based jogging program of 120 min/wk at 75% VO2max. Only 12 men showed a sufficient exercise adherence over the 6 months (> or = 60 min/wk on average) and were included into statistical analysis. Their average training activity was 105 +/- 31 min/wk. The results revealed significant increases in VO2max (+8.4%, p < 0.01) and submaximal power output (+18.1%, p < 0.01). Total mitochondrial volume density in M. vastus lateralis increased by 20% (p < 0.05) with a larger increase in subsarcolemmal volume compared to central volume (50% vs 15%). No changes in volume of intracellular lipid droplets, capillary density, capillary per fiber ratio, fiber mean cross-sectional area and muscle fiber type could be observed. Body composition analysis showed a decrease in trunk fat mass (-7.3%, p < 0.05) and an increase in trunk lean mass (+1.7%, p < 0.05), while changes in the legs were not significant. It can be concluded that a moderate-intensity, health oriented endurance training beneficially affected cardiovascular and muscle oxidation capacity as well as body composition in the trunk area. No adaptations in capillaries or lipid metabolism could be demonstrated. The results support the hypothesis of thresholds for induction of adaptation processes in muscle skeletal tissue depending on the intensity of the exercise stimulus.

Adipose Tissue↗
PubMed1995

mRNAs of enzymes involved in energy metabolism and mtDNA are increased in endurance-trained athletes.

Improvements in endurance capacity by training are associated with structural and biochemical adaptations of working muscles that affect the mitochondrial compartment. We investigated whether the 1.8-fold higher mitochondrial volume density in a group of endurance-trained athletes compared with untrained subjects was reflected by higher steady-state levels of mRNAs coding for components of the oxidative phosphorylation pathway using a quantitative polymerase chain reaction approach. We found that mitochondrially encoded RNAs (cytochrome-c oxidase subunit I, NADH reductase subunit 6, 16S rRNA), as well as nuclear-encoded RNAs (cytochrome-c oxidase subunit IV, succinate dehydrogenase, fumarase) are all increased coordinately in the athletes (1.54- to 1.94-fold). In addition, mitochondrial (mt) DNA concentration was also 1.55-fold higher in the trained athletes, whereas genomic DNA was not changed. Our findings thus show similar RNA expression of mitochondrially encoded genes in sedentary and endurance-trained subjects, whereas pretranslational control mechanisms account for higher levels of nuclear-encoded RNAs in the athletes.

Adult↗
PubMed1994

An efficient polymerase chain reaction approach for the quantitation of multiple RNAs in human tissue samples.

We describe a PCR quantitation approach that we have set up to study gene expression in human skeletal muscle biopsies. It is characterized by the independent standardization of the individual steps and ignores attempts to control for the efficiency of the PCR. Different RNA extraction/reverse transcription efficiencies are normalized by the addition of an unrelated cRNA. Quantitation is achieved by parallel amplifications of reference samples containing known amounts of PCR products. Precision was achieved by multiple measurements of several samples. Our approach allows for the detection of less than twofold differences (27-88%, depending on the RNA species studied) among samples. A comparison of biopsies from highly trained endurance runners with biopsies from untrained subjects showed that the increased mitochondrial density in the runners' samples is accompanied by a proportional increase in the concentration of the mRNA of cytochrome c oxidase subunit IV.

Adult↗
PubMed1994

The use of 33P-labelled riboprobes for in situ hybridizations: localization of myosin alkali light-chain mRNAs in adult human skeletal muscle.

33P-labelled probes were used to localize the mRNAs coding for the myosin alkali light-chain isoforms MLC 1f/3f and MLC 1sb in adult human muscles, which are distributed in characteristic fibre type specific patterns. In situ hybridizations of 33P-labelled probes were compared with probes carrying 35S or digoxigenin labels. Signals of equal strength were obtained with each of the three labels. The preferentially peripheral localization of these mRNAs in the muscle fibres could be clearly seen with all three probes, with digoxigenin probes providing the best resolution. 33P can serve as a viable alternative in this type of experiment. These experiments with adult human muscles also showed that the post mortem stability of RNA in human muscle is better than generally assumed. We could detect no signs of degradation in RNA prepared from heart ventricle as well as skeletal muscle up to 24 hours post mortem. In situ hybridizations worked equally well in biopsy material as in post mortem samples.

Adult↗
PubMed1994

[Basis of muscle contraction].

This interdisciplinary review aims at updating the knowledge on structural and functional properties of skeletal muscle tissue to the current state of art in various disciplines of basic research on muscle tissue. The mechanism of contraction as well as the excitation-contraction coupling are discussed along with recent findings on the molecular organization of muscle filaments and further constituents of the cytoskeleton. The structural conditions for oxygen and substrate metabolism prevailing in the working muscle cell are outlined and discussed with respect to the fiber type composition and recruitment of muscles during athletic activities.

Calcium↗
PubMed1992

Fast myosin light chain expression in chicken muscles studied by in situ hybridization.

We have studied the fiber type-specific expression of the fast myosin light chain isoforms LC 1f, LC 2f, and LC 3f in adult chicken muscles using in situ hybridization and two-dimensional gel electrophoresis. Type II (fast) fibers contain all three fast myosin light chain mRNAs; Types I and III (slow) fibers lack them. The myosin light chain patterns of two-dimensional gels from microdissected single fibers match their mRNA signals in the in situ hybridizations. The results confirm and extend previous studies on the fiber type-specific distribution of myosin light chains in chicken muscles which used specific antibodies. The quantitative ratios between protein and mRNA content were not the same for all three fast myosin light chains, however. In bulk muscle samples, as well as in single fibers, there was proportionally less LC 3f accumulated for a given mRNA concentration than LC 1f or LC 2f. Moreover, the ratio between LC 3f mRNA and protein was different in samples from muscles, indicating that LC 3f is regulated somewhat differently than LC 1f and LC 2f. In contrast to other in situ hybridization studies on the fiber type-specific localization of muscle protein mRNAs, which reported the RNAs to be located preferentially at the periphery of the fibers, we found all three fast myosin light chain mRNAs quite evenly distributed within the fiber's cross-sections, and also in the few rare fibers which showed hybridization signals several-fold higher than their surrounding counterparts. This could indicate principal differences in the intracellular localization among the mRNAs coding for various myofibrillar protein families.

Animals↗
PubMed1991

Conditions for oxygen and substrate transport in muscles in exercising mammals.

The structural conditions relevant for metabolite exchange in anaerobic and aerobic work conditions in muscle tissue are reviewed. High-intensity non-steady-state exercise is supported by the phosphocreatine pool, which serves as a shuttle for high-energy phosphates produced by glycolysis and by aerobic metabolism. This is achieved through the intermediary of a topologically organized creatine kinase isozyme system. The muscle capillary network supplies substrate and environmental oxygen to the mitochondria. The network is quantitatively matched to the muscle oxidative capacity, determined structurally by mitochondrial volume. Capillary hematocrit, erythrocyte spacing and oxygen saturation of myoglobin are critical variables for oxygen release from microvessels. Myoglobin greatly helps intracellular oxygen transfer as, under aerobic work conditions, it keeps intracellular oxygen tension low and uniform in the muscle fibers. During sustained submaximal work, muscle cells are fueled by both endogenous (triglycerides and glycogen) and circulatory (lactate, glucose and fatty acids) substrates. A lactate shuttle in which lactate may move through the circulation, as well as directly from fiber to fiber, provides many of the carbohydrate-derived carbon skeletons for terminal oxidation. Glucose is taken up from the interstitial space by facilitated diffusion, mostly mediated by a glucose transporter (GLUT4) that is translocated from an intracellular location to the sarcolemma by activity and insulin. Extramyocellular transport of fatty acids is mediated by albumin, while fatty-acid-binding proteins are held responsible for intracellular fatty acid transport.

Animals↗
PubMed1988

Approximately 1 kilobase of sequence 5' to the two myosin light-chain 1f/3f gene cap sites is sufficient for differentiation-dependent expression.

Approximately 1 kilobase of genomic DNA from the chicken fast myosin light-chain 1f/3f gene 5' to the transcriptional start sites for each light-chain mRNA was sufficient for differentiation-dependent, tissue-restricted expression. This was determined in primary chick myoblast cultures transfected with the chloramphenicol acetyltransferase (CAT) expression vector p8CAT containing these 5'-flanking sequences. The expression of CAT activity from both light-chain promoters was 10- to 20-fold higher in differentiated myotubes than in fibroblasts or myoblasts grown in bromodeoxyuridine. In contrast, the beta-actin and Rous sarcoma virus promoters joined to the CAT gene were expressed equally in all cell backgrounds tested. Even though the relative timing of light-chain 1f and 3f expression was altered, tissue-restricted, differentiation-dependent expression of the light-chain mRNAs was maintained with these 5' cis-acting sequence elements.

Acetyltransferases↗
PubMed1984

Heterogeneous distribution of myosin in human masticatory muscle fibres as shown by immunocytochemistry.

On the basis of enzymic properties, different fibre types can be distinguished in human skeletal muscle (type I fibres and type II fibres with subtypes) and there is a correlation between fibre types and the occurrence of slow and fast myosin. In human masticatory muscles, fibres with ATPase activity at pH 9.4, intermediate between that of type I (low activity) and type II (high activity), are frequent. On cryostat-sectioned material, highly specific antibodies against fast myosin, slow myosin and slow light chains were applied. The myosin composition of human masticatory muscles was very heterogeneous, in contrast to that in limb muscles, with various proportions of slow and fast myosins, heavy as well as light chains. Type I fibres contained slow myosin only and type II mainly fast myosin, ATPase IM and type IIC fibres contained a mixture of slow and fast myosins in variable amounts. The findings conform with physiological evidence of a continuum of contraction times for motor units in the human masticatory muscles and suggests that these muscles are highly adapted to the special and complicated functions of the stomatognathic system.

Adenosine Triphosphatases↗
PubMed1984

Muscle fibre type composition in distal myopathy (Welander). An analysis with enzyme- and immuno-histochemical, gel-electrophoretic and ultrastructural techniques.

The myopathic muscle of distal myopathy (Welander's disease), the dominantly inherited neuromuscular disorder which occurs frequently in Sweden, has been characterized by electron microscopy, enzyme- and immuno-histochemistry (using antibodies against embryonic, neonatal, fast and slow myosin, and against the muscle-specific intermediate filament protein, desmin), and with gel electrophoretic techniques. Of special interest is the fact that the ultrastructural appearance of the fibres with regard to M- and Z-band structures does not fit the proposed classification criteria for ultrastructural fibre typing of normal human muscle. Furthermore, contrary to previous results, we conclusively demonstrate that the predominating fibres are of a slow-twitch type. Unexpectedly, we also observed that embryonic and neonatal myosin was expressed in some residual fibres. This emphasises the importance of supplementing stains to demonstrate activity of ATPase with myosin immuno-histochemistry in order to improve understanding of fibre type characteristics in myopathic muscles. The origin of the myopathic muscle fibres in distal myopathy could not be definitely determined, but it is suggested that neurogenic disturbances play an important part in the pathophysiology of Welander's disease.

Adenosine Triphosphatases↗
PubMed1984

Development of fiber types in human fetal muscle. An immunocytochemical study.

Human fetal muscles have been studied using immunocytochemical methods with antibodies directed against different myosin isoforms. We show that fiber type differences can be detected as early as 15-16 weeks of gestation. At this time it would appear that both the heavy and light chains characteristic of slow myosin are found in some myotubes.

Adenosine Triphosphatases↗
PubMed1983

A new myofibrillar protein characteristic of type I human skeletal muscle fibres.

Analysis of a single human type I (slow-twitch) skeletal muscle fibres by two-dimensional gel electrophoresis shows that they contain a specific protein of subunit Mr 34 x 10(3) and pI 6.0, 6.1 and 6.2 (three spots) which is absent from type IIA and IIB muscle fibres. This type I protein is located in the I-band of isolated myofibrils as demonstrated by the indirect immunofluorescence using specific antisera against the denatured form of type I protein. Type I protein could only be extracted from myofibrils with high salt solutions (0.6-1 M KCl), conditions which also coextract actin and myosin, demonstrating its firm attachment. Rat and cat type I muscle fibres as well as the slow-tonic fibres from chicken anterior latissimus dorsi also contained a protein homologous to the human type I protein. The human type I protein was found to be different from the known regulatory and contractile muscle proteins by electrophoretic and immunological criteria, and may therefore be a new myofibrillar protein in type I muscle fibres.

Animals↗
PubMed1982

Training-induced increase in myofibrillar ATPase intermediate fibers in human skeletal muscle.

The response of human muscles triceps brachii and vastus lateralis to training, consisting of 18.5 miles a day of skiing with a backpack, 6 days a week for 8 weeks, has been investigated by means of histochemical fiber typing (myofibrillar ATPase) as well as immunohistochemical discrimination of slow and fast myosin (n = 6). A detraining period of 33 weeks was also studied. The percentage of type 2 fibers decreased by 6% in the triceps brachii during training, whereas the intermediate fiber type increased by 4%. No change was found in the distribution of fiber types in the vastus lateralis during training or in both muscles during detraining. The ATPase intermediate fibers also stained intermediately in the immunohistochemical stainings for slow and fast myosin. These results show that transformation of a fraction of the type 2 fibers into the intermediate type may occur as a response to endurance training.

Adenosine Triphosphatases↗
PubMed1982

No classical type IIB fibres in dog skeletal muscle.

To analyse the fibre type composition of adult dog skeletal muscle, enzyme histochemistry, immunohistochemistry for type I, IIA and IIB myosins, and peptide mapping of myosin heavy chains isolated from typed single according to the activity of the m-ATPase after acidic and alkaline preincubation proved to be rather difficult and was only consistently achieved after a very careful adjustment of the systems used. One of these sub-classes of type II fibres stained more strongly for m-ATPase activity after acidic and alkaline preincubation, was oxidative-glycolytic and showed a strong reaction with an anti-type IIA myosin. The other one, however, although showed a faint reaction with an anti-type IIB myosin. Peptide mapping of the myosin heavy chains of typed single fibres revealed two populations of heavy chains among the type II fibre group. Thus, in dog muscle, we are confronted with the presence of two main classes of type II fibres, both oxidative-glycolytic, but differing in the structure of their myosin heavy chains. In contrast to some reports in the literature, no classical type IIB fibres could be detected.

Adenosine Triphosphatases↗