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Biomedical subjects

D Pette

Publications and source records attributed to D Pette.

At least 181 records · Page 10Linked to original sources
PubMed1987

Myosin polymorphism in single fibers of chronically stimulated rabbit fast-twitch muscle.

Rabbit tibialis anterior (TA) muscles were indirectly stimulated (10 Hz, 24 h/d) for 30 d and 60 d and single fibers were analysed using a combined histochemical and biochemical technique (Staron and Pette 1986, 1987a, b). After 30 d of chronic stimulation there was a pronounced increase in the normally rare (0.5%) C fiber population (i.e., fibers containing slow- and fast-myosins in varying ratios). At this time, C fibers amounted to almost 60% of the total population. In the 60 d stimulated muscles, the major population (98%) consisted of an atypical type It fiber. This fiber type which was not detectable in normal TA muscle, differed histochemically and biochemically from type I fibers. It contained the slow-myosin light chains LC1s and LC2s, the heavy chain HCI, and, in addition, high concentrations of the fast-myosin alkali light chain LC1f and possibly traces of a heavy chain with an electrophoretic mobility comparable with that of the fast-myosin heavy chain HCIIa. These It fibers were occasionally observed in the unstimulated, contralateral TA muscles which also contained an increased population of C fibers (1.3-6.3%). Although the transformation even after 60 d of chronic stimulation was incomplete, these changes demonstrate the ability of muscle fibers to adapt in a specific manner to altered functional demands brought about by an altered stimulus pattern. In addition, the pronounced heterogeneity of the fiber population undergoing transformation indicates a nonuniform response to a uniform stimulus pattern.

Animals↗
PubMed1987

Developmental changes in succinate dehydrogenase activity in muscle fibers from normal and dystrophic mice.

The question of whether or not the development of dystrophic muscles is similar to that of normal muscles, prior to the manifestations of the symptoms of the disease, is investigated here. The developmental change in the activity of succinate dehydrogenase was therefore measured in individual fibers of prospectively dystrophic muscles from 10- to 28-day-old mice (strain C57Bl/6J dy2j) and compared with that of muscles from normal mice of the same age. It was found that up to 10 days of age, muscle fibers from normal and prospective dystrophic animals had low succinate dehydrogenase activities, and were all more or less uniform. Thereafter in the normal muscle the overall activity of the enzyme increased and the fibers became more heterogeneous with age. By 21 days the extensor digitorum longus muscle resembled that of the adult. At that time, fibers from prospectively dystrophic muscles had lower succinate dehydrogenase activities and were more homogeneous. Thus fibers from prospectively dystrophic muscles fail to achieve their adult characteristics by 21 days. On the basis of these results, it is suggested that muscle maturation is retarded in dystrophic animals.

Animals↗
PubMed1986

Neural control of gene expression in skeletal muscle. Calcium-sequestering proteins in developing and chronically stimulated rabbit skeletal muscles.

Tissue contents of the sarcoplasmic-reticulum Ca2+-ATPase (Ca2+ +Mg2+-dependent ATPase), of calsequestrin and of parvalbumin were immunochemically quantified in homogenates of fast- and slow-twitch muscles of embryonic, maturing and adult rabbits. Unlike parvalbumin, Ca2+-ATPase and calsequestrin were expressed in embryonic muscles. Presumptive fast-twitch muscles displayed higher contents of these two proteins than did presumptive slow-twitch muscles. Calsequestrin steeply increased before birth and reached adult values in the two muscle types 4 days after birth. The main increase in Ca2+-ATPase occurred during the first 2 weeks after birth. Denervation of postnatal fast- and slow-twitch muscles decreased calsequestrin to amounts typical of embryonic muscle and suppressed further increases of Ca2+-ATPase. Denervation caused slight decreases in Ca2+-ATPase in adult fast-twitch, but not in slow-twitch, muscles, whereas calsequestrin was greatly decreased in both. Chronic low-frequency stimulation induced a rapid decrease in parvalbumin in fast-twitch muscle, which was preceded by a drastic decrease in the amount of its polyadenylated RNA translatable in vitro. Tissue amounts of Ca2+-ATPase and calsequestrin were essentially unaltered up to periods of 52 days stimulation. These results indicate that in fast- and slow-twitch muscles different basal amounts of Ca2+-ATPase and calsequestrin are expressed independent of innervation, but that neuromuscular activity has a modulatory effect. Conversely, the expression of parvalbumin is greatly enhanced by phasic, and drastically decreased by tonic, motor-neuron activity.

Animals↗
PubMed1986

Neural control of gene expression in skeletal muscle. Effects of chronic stimulation on lactate dehydrogenase isoenzymes and citrate synthase.

The aim of this study was to investigate the effects of neural activity on the expression of fibre-type-specific patterns of metabolic enzymes at the levels of transcription and translation. For this purpose, changes in tissue amounts of citrate synthase (CS) and the H- and M-subunits of lactate dehydrogenase (LDH) were followed in fast-twitch rabbit muscles during low-frequency (10 Hz, 12 h/day) nerve stimulation. These stimulation-induced alterations were correlated with changes in tissue amounts of the total poly(A)+ (polyadenylated) RNA, poly(A)+ RNAs specifically translatable in vitro, yield of total ribosomes and distributions of monosomes and polysomes. The tissue contents of poly(A)+ RNAs translatable in vitro coding for CS and H- and M-LDH were quantified by immunoprecipitation of their translation products. Increases in total ribosome yields occurred after 4 days' stimulation, reaching a maximum between 14 and 21 days. Stimulation for only 1-2 days greatly increased the amount of monosomes. An increase in polysomes occurred before that in total ribosomes, suggesting that monosomes were integrated into polysomes. Total poly(A)+ RNA significantly increased in muscles stimulated for more than 6 days. A maximum increase of 2.5-fold was attained after 14-21 days. Chronic stimulation progressively induced the appearance of LDH isoenzymes containing the H-subunit, with a predominance of LDH-3. This shift corresponded to a slow decay of the M-subunit and a 2-fold steep increase in the H-subunit. These changes correlated with those of the respective poly(A)+ RNAs translatable in vitro, thus indicating that the re-arrangement of the LDH isoenzyme pattern is mainly due to qualitatively and quantitatively altered transcription. The increase in CS was biphasic and consisted of a moderate rise during the first 4 days of stimulation and a steep rise thereafter. The latter coincided with a steep increase in poly(A)+ RNA translatable in vitro coding for CS. In view of the early increase in translational capacity, it was concluded that the initial rise in CS resulted from selective post-transcriptional control and enhanced translation in vivo of existing mRNA, whereas its steep increase was due to enhanced transcription. These results indicate that the neurally regulated expression of phenotype-specific properties in muscle includes control of both transcription and translation.

Animals↗
PubMed1986

Decrease in myosin light chain kinase activity of rabbit fast muscle by chronic stimulation.

Analysis of myosin light chain kinase (MLCK) activity in tibialis anterior muscles of the rabbit revealed that chronic stimulation at a frequency of 10 Hz for 24 h per day reduced the enzyme activity in a time-dependent manner. Since fast twitch muscle contains significantly more myosin light chain kinase than slow twitch muscle, the observed reductions are consistent with the type of fast-to-slow transformation observed for other type-specific muscle characteristics. The present data also indicate that the stimulation-induced decrease in MLCK activity precedes the fast-to-slow conversion of the myosin molecule as judged by pyrophosphate-polyacrylamide gel electrophoresis.

Animals↗
PubMed1986

Immunochemical quantification of sarcoplasmic reticulum Ca-ATPase, of calsequestrin and of parvalbumin in rabbit skeletal muscles of defined fiber composition.

Antibodies directed against purified Ca-ATPase from sarcoplasmic reticulum, calsequestrin and parvalbumin from rabbit fast-twitch muscle were raised in sheep. The specificity of the antibodies was shown by immunoblot analysis and by enzyme-linked immunoadsorbent assays (ELISAs). IgG against the sarcoplasmic reticulum Ca-ATPase inhibited the catalytic activities of Ca-ATPase from fast-twitch (psoas, tibialis anterior) and slow-twitch (soleus) muscles to the same degree. In non-equilibrium competitive ELISAs the anti(Ca-ATPase) IgG displayed a slightly higher affinity for the Ca-ATPase from fast-twitch muscle than for that from slow-twitch muscle. This suggests a fiber-type-specific polymorphism of the sarcoplasmic reticulum Ca-ATPase. Quantification of Ca-ATPase, calsequestrin and parvalbumin in various rabbit skeletal muscles of histochemically determined fiber composition was achieved by sandwich ELISA. Ca-ATPase was found to be 6-7 times higher in fast than in slow-twitch muscles. A slightly higher concentration was found in fast-twitch muscles with a higher percentage of IIb fibers when compared with fast-twitch muscles with a higher percentage of IIa fibers. Thus Ca-ATPase is distributed as follows, IIb greater than or equal to IIa much greater than I. Calsequestrin was uniformly distributed in fast-twitch muscles independently of their IIa/IIb fiber ratio and displayed 50% lower concentrations in slow than in fast-twitch muscles (IIb = IIa greater than I). Parvalbumin contents were 200-300-fold higher in fast than in slow-twitch muscles. Significantly lower parvalbumin concentrations were found in fast-twitch muscles with a higher percentage of IIa fibers than in fast-twitch muscles with a higher percentage of IIb fibers (IIb greater than IIa much greater than I).

Animals↗
PubMed1986

Neural regulation of parvalbumin expression in mammalian skeletal muscle.

Parvalbumin was purified from rabbit fast skeletal muscle and used to raise antibodies in sheep. Subsequently, a sensitive 'sandwich' enzyme-linked immunoadsorbent assay permitted quantification of parvalbumin in homogenates of embryonic, maturing, innervated, denervated and chronically stimulated skeletal muscles of the rabbit. High concentrations of parvalbumin were detected in various adult fast-twitch muscles of the rabbit (700-1200 micrograms/g of muscle), whereas slow-twitch muscles contained negligible concentrations (3-5 micrograms/g of muscle). Parvalbumin was not detectable in embryonic-rabbit muscles (21, 25, 28 days of gestation), either presumptive fast- or slow-twitch. However, parvalbumin concentrations did increase during postnatal development in presumptive fast-twitch muscles. Thus the onset of parvalbumin synthesis appears to be correlated with the neonatal-to-adult transition of motor-neuron activity [Navarrete & Vrbová (1983) Dev. Brain Res. 8, 11-19]. The increase of parvalbumin in maturing, presumptive fast-twitch muscle was suppressed by denervation. In the adult rabbit, denervation of the tibialis anterior muscle caused a reduction of parvalbumin to a level normally found in slow-twitch muscles. In contrast, the already low levels of parvalbumin in maturing and adult slow-twitch soleus muscle were unaffected by denervation. Chronic low-frequency stimulation of adult fast-twitch muscle resulted in a rapid reduction of parvalbumin to a level normally found in slow-twitch muscle. These data support the hypothesis that the expression of parvalbumin is under positive control of fast-type motor-neuron activity.

Animals↗
PubMed1986

Degeneration-regeneration as a mechanism contributing to the fast to slow conversion of chronically stimulated fast-twitch rabbit muscle.

Extensor digitorum longus muscles of male adult White New Zealand rabbits were indirectly stimulated at 10 Hz for 12 h daily for periods ranging up to 28 days. After four weeks the stimulated muscles showed a nearly uniform profile of high succinate dehydrogenase activity and, when incubated after acid preincubation for myofibrillar adenosine triphosphatase, displayed more dark- and intermediate-staining fibers than their contralateral counterparts. Muscles stimulated from between 6 to 21 days revealed degenerative foci and phagocytosis of degenerated fibers. These fibers were mostly of the fast-twitch, glycolytic type. Small myofibers, which often contained central nuclei, and structures identified as myoblasts or myotubes, reacted with a monoclonal antibody prepared against embryonic myosin heavy chains. The data suggest that under the employed conditions the fast to slow conversion of chronically stimulated fast-twitch rabbit muscle is not exclusively caused by adult fiber transformation, but results in part from the substitution of fast-twitch glycolytic fibers with newly formed fibers that have a high oxidative profile.

Adenosine Triphosphatases↗
PubMed1986

Correlation between myofibrillar ATPase activity and myosin heavy chain composition in rabbit muscle fibers.

Combined histochemical and biochemical analyses were performed on single fibers of rabbit soleus muscle. Histochemically, four fiber types (I, IC, IIC, IIA) were defined. Of these, types I and IIA were separate, histochemically homogeneous groups. A heterogeneous C fiber population exhibited a continuum of staining intensities between types I and IIA. Microelectrophoretic analyses of specific, histochemically defined fibers revealed that type I fibers contained exclusively HCI, whereas type IIA fibers contained only HCIIa. The C fibers were characterized by the coexistence of both heavy chains in varying ratios, type IC with a predominance of HCI and type IIC with a predominance of HCIIa. A direct correlation existed between the myosin heavy chain composition and the histochemical mATPase staining and was especially evident in the C fiber population with its variable HCI/HCIIa ratio. This correlation did not apply to the myosin light chain complement.

Adenosine Triphosphatases↗
PubMed1986

Distribution of sarcoplasmic reticulum Ca-ATPase and of calsequestrin in rabbit and rat skeletal muscle fibers.

Muscle fibers in rabbit extensor digitorum longus (EDL), tibialis anterior (TA) and soleus, and rat soleus, were examined immunohistochemically for two proteins of the sarcoplasmic reticulum. Ca-ATPase and calsequestrin (CaS). Fibers were typed with the histochemical reaction for actomyosin ATPase. In the rabbit EDL and TA, type I fibers clearly reacted less for Ca-ATPase and CaS than type II fibers, but the difference was less with CaS than with Ca-ATPase. Although the differences were relatively small, IIB fibers consistently presented greater amounts of Ca-ATPase than IIA fibers. No type II subgroups could be recognized after incubation with anti-CaS. These findings confirm results from previous immunochemical measurements on whole muscles containing different proportions of IIA and IIB fibers (Leberer and Pette 1986). Type IIA and IIC in the rabbit and rat soleus reacted stronger for Ca-ATPase and for CaS than type I fibers. Small differences in Ca-ATPase, but not in CaS, were recognized within the type I fiber population. Therefore, type I fibers in the rabbit and rat soleus are not a homogeneous population.

Animals↗
PubMed1986

Actin-severing activity copurifies with phosphofructokinase.

Microinjection of muscle 6-phosphofructokinase (PFK; EC 2.7.1.11) into tissue culture cells led to a reversible disintegration of microfilament bundles (stress fibers). The mode of disruption as well as of recovery of stress fibers was very similar to that found previously in experiments performed with the actin-severing protein brevin, an extracellular variant of gelsolin. PFK, like brevin, was also capable of disrupting stress fibers in detergent-extracted cells and in ethanol-fixed cells, in a Ca2+-dependent manner. When compared with heart muscle gelsolin, PFK comigrated with the 85- to 90-kDa band. Antibodies against PFK crossreacted with gelsolin from the same species. These results point to a tight association between polypeptides with similar biochemical and immunological parameters present in both preparations. They suggest hitherto unexpected cellular control mechanisms for both microfilament functions and glycolysis.

Actin Cytoskeleton↗
PubMed1986

Enzyme activities in single electrophysiologically identified crab muscle fibres.

The superficial muscle fibres in the proximal part of the closer muscle in the crab Eriphia can be separated into four fibre groups (I-IV) on the basis of electrophysiological and histochemical characteristics. The activity levels of glyceraldehydephosphate dehydrogenase (GAPDH), lactate dehydrogenase (LDH), citrate synthase (CS), NADP-isocitrate dehydrogenase (IDH) and 3-hydroxyacyl-CoA dehydrogenase (HAD), determined in single electrophysiologically identified fibres, differed significantly among the different fibre groups. In addition, fibres belonging to the same group, with similar electrophysiological characteristics, demonstrated variability with regard to metabolic enzyme activities. Nevertheless, comparison of absolute enzyme activities and enzyme activity ratios permitted the discrimination of at least three groups. These groups corresponded with those defined according to electrophysiological and histochemical characteristics. The group I fibres (tonic fibres) are intermediate in oxidative potential and show the lowest values of glycolytic enzymes. The group II and group III fibres can be regarded as fast oxidative fibres. The high ratio between activity levels of enzymes for glycolytic and oxidative metabolism found for group IV fibres (fast fibres) demonstrated that this group depends strongly on anaerobic metabolism.

3-Hydroxyacyl CoA Dehydrogenases↗
PubMed1986

Metabolic properties of muscle fibers.

Mammalian skeletal muscles are composed of slow (type I) and fast (type II) twitch fibers, which, as reflected by their enzyme activity patterns, are characterized by specific metabolic properties. Type I fibers are always "oxidative" but nevertheless form a spectrum. Type II fibers likewise form a spectrum but display a wider range with "oxidative" and "glycolytic" extremes. As a result, type I and type II fibers can be classified independently of myofibrillar ATPase histochemistry by their specific enzyme activity profiles. In this context, activity ratios between enzymes of anaerobic and aerobic pathways can be used as discriminative parameters. Similarly, specific ratios of enzymes catalyzing unidirectional reactions in hexose metabolism (hexokinase, phosphofructokinase, fructose-1,6-bisphosphatase) separate the two fiber populations. The histochemically defined IIA and IIB subtypes cannot be separated into distinct metabolic groups. In view of the continuum of metabolic properties, skeletal muscle is an extremely heterogeneous tissue in which each fiber represents a separate metabolic compartment.

Aerobiosis↗
PubMed1985

Increased mitochondrial creatine kinase in chronically stimulated fast-twitch rabbit muscle.

Fractional extraction and isozyme electrophoresis revealed the presence of small amounts (2.5% of total cellular activity) of mitochondrial creatine kinase (CK) in rabbit fast-twitch muscle. Chronic nerve stimulation resulted in a decrease of extramitochondrial MM-CK to 60% of its normal value but induced an approx. 4-fold increase in mitochondrial CK. This increase occurred in parallel with the rise in enzyme activities of terminal substrate oxidation.

Animals↗
PubMed1985

Decreased parvalbumin contents in skeletal muscles of C57BL/6J(dy2J/dy2J) dystrophic mice.

Parvalbumin content is decreased by 40% in hindlimb muscles of adult (3.5 months) dystrophic mice when compared to normal mice of the same age. The concentration of parvalbumin in the dystrophic muscles resembles that of immature muscles of 4-week-old normal mice. The reduction may contribute to an elevation of sarcoplasmic free CA2+, which could stimulate various Ca2+-dependent processes related to the disease.

Animals↗
PubMed1985

Neural control of phenotypic expression in mammalian muscle fibers.

In this review, the present knowledge about the mechanisms involved in the control of the phenotypic expression of mammalian muscle fibers is summarized. There is a discussion as to how the activity imposed on the muscle fibers by the motoneuron finally induces in the muscle cells the expression of those genes that define its particular phenotype. The functional and molecular heterogeneity of skeletal muscle is thus defined by the existence of motor units with varied function, while the homogeneity of muscle fibers belonging to the same motor unit is yet another indication of the importance of activity in the control of gene expression of the mammalian muscle fiber.

Animals↗