PubMed Health⌕ Search

Biomedical subjects

G Vrbova

Publications and source records attributed to G Vrbova.

At least 19 recordsLinked to original sources
PubMed2001

Deficiency in parvalbumin increases fatigue resistance in fast-twitch muscle and upregulates mitochondria.

The soluble Ca2+-binding protein parvalbumin (PV) is expressed at high levels in fast-twitch muscles of mice. Deficiency of PV in knockout mice (PV -/-) slows down the speed of twitch relaxation, while maximum force generated during tetanic contraction is unaltered. We observed that PV-deficient fast-twitch muscles were significantly more resistant to fatigue than were the wild type. Thus components involved in Ca2+ homeostasis during the contraction-relaxation cycle were analyzed. No upregulation of another cytosolic Ca2+-binding protein was found. Mitochondria are thought to play a physiological role during muscle relaxation and were thus analyzed. The fractional volume of mitochondria in the fast-twitch muscle extensor digitorum longus (EDL) was almost doubled in PV -/- mice, and this was reflected in an increase of cytochrome c oxidase. A faster removal of intracellular Ca2+ concentration ([Ca2+]i) 200-700 ms after fast-twitch muscle stimulation observed in PV -/- muscles supports the role for mitochondria in late [Ca2+]i removal. The present results also show a significant increase of the density of capillaries in EDL muscles of PV -/- mice. Thus alterations in the dynamics of Ca2+ transients detected in fast-twitch muscles of PV -/- mice might be linked to the increase in mitochondria volume and capillary density, which contribute to the greater fatigue resistance of these muscles.

Animals↗
PubMed1999

Prolonged contraction-relaxation cycle of fast-twitch muscles in parvalbumin knockout mice.

The calcium-binding protein parvalbumin (PV) occurs at high concentrations in fast-contracting vertebrate muscle fibers. Its putative role in facilitating the rapid relaxation of mammalian fast-twitch muscle fibers by acting as a temporary buffer for Ca2+ is still controversial. We generated knockout mice for PV (PV -/-) and compared the Ca2+ transients and the dynamics of contraction of their muscles with those from heterozygous (PV +/-) and wild-type (WT) mice. In the muscles of PV-deficient mice, the decay of intracellular Ca2+ concentration ([Ca2+]i) after 20-ms stimulation was slower compared with WT mice and led to a prolongation of the time required to attain peak twitch tension and to an extension of the half-relaxation time. The integral [Ca2+]i in muscle fibers of PV -/- mice was higher and consequently the force generated during a single twitch was approximately 40% greater than in PV +/- and WT animals. Acceleration of the contraction-relaxation cycle of fast-twitch muscle fibers by PV may confer an advantage in the performance of rapid, phasic movements.

Animals↗
PubMed1998

Myosin regulatory elements as vectors for gene transfer by intramuscular injection.

Intramuscular injection of plasmid constructs promises to be an effective way of carrying out gene therapy for muscle disorders as well as using muscle as an in vivo expression system for disorders that involve the gene product being secreted into the bloodstream. The effectiveness of this method depends on the design of the cassette used for the expression of the cDNA of the introduced gene. We tested the levels of expression achieved by a number of muscle-specific promoters and a myosin light chain enhancer when spliced to the reporter gene chloramphenicol acetyltransferase (CAT), in vitro and in vivo by injection into fast and slow muscles of the mouse. The results show that the highest levels of expression are achieved by a combination of a truncated myosin heavy chain promoter and the enhancer, and that a whole range of expression levels is obtained with the other combinations tested. The data show that a cassette based on these elements should provide efficient vectors for the introduction and expression of genes following intramuscular injection of naked DNA.

Animals↗
PubMed1996

Effect of fatiguing maximal voluntary contraction on excitatory and inhibitory responses elicited by transcranial magnetic motor cortex stimulation.

Vertex transcranial magnetic stimulation (TMS) elicited tibialis anterior motor evoked potentials (MEPs) and silent periods (SPs) that were recorded during and following isometric maximal volitional contraction (MVC). During MVC in 6 healthy subjects, MEP amplitudes in the exercised muscle showed an increasing trend from an initial value of 4539 +/- 809 muV (mean +/- SE) to 550 +/- 908 muV (P < 0.13) while force and EMG decreased (P < 0.01). Also, SP duration increased from 165 +/- 37 ms to 231 +/- 32 ms (P < 0.01). Thus, during a fatiguing MVC both excitatory and inhibitory TMS-induced responses increased. TMS delivered during repeated brief 10% MVC contractions before and after a fatiguing MVC in 5 subjects, showed no change in MEP amplitude but SP duration was prolonged after MVC. This SP prolongation was focal to the exercised muscle. Silent periods recorded after pyramidal tract stimulation were unchanged following the MVC. These results suggest that MEP and SP might have common sources of facilitation during an MVC and that inhibitory mechanisms remain focally augmented following a fatiguing MVC.

Adult↗
PubMed1995

Motor activity patterns in rat soleus muscle after neonatal partial denervation.

In normal rats the development of organized patterns of hind limb movements takes place during the first three weeks of life. After removal of a part of the rat soleus muscle's innervation in 5-day-old animals, the remaining motoneurones occupy a large peripheral field. The possibility that the development of the normal activity patterns of these motor units may be altered was studied. The EMG activity of the soleus muscles partially denervated at five days was compared to that of the contralateral unoperated muscles during spontaneous locomotion and induced reflex activity in animals at various ages. Like a normal soleus the partially denervated soleus developed with age a tonic activity pattern but the aggregate activity recorded from the partially denervated soleus was less than that in the control muscle. However, the amount of activity per motor unit was higher in the operated than in the control muscles, since these had only one-third to half of their normal complement of motor units. During locomotion both soleus muscles were activated like typical ankle extensors during the stance phase of the step cycle, but the burst duration of the operated muscle was significantly shorter. We conclude that partial denervation shortly after birth leads to an overall increase in activity of the remaining soleus motor units but does not drastically alter their temporal pattern of use during locomotion.

Animals↗
PubMed1995

Blocking of NMDA receptors during a critical stage of development reduces the effects of nerve injury at birth on muscles and motoneurones.

Blocking of NMDA receptors during a critical stage of development reduces the effects of nerve injury at birth on muscles and motoneurones. Injury to the sciatic nerve at birth causes many motoneurones to soleus and extensor digitorum longus (EDL) muscles of rats to die. This is reflected in a reduction of motor units in these muscles. In the soleus only 4 (12.3%) motor units remain while 10 (24.3%) remain in the EDL, showing that soleus alpha motoneurones are more sensitive to nerve injury at birth. Treatment with MK-801, an NMDA receptor blocker, rescues a proportion of motor units in both muscles, so that in the soleus 11 (36%) and in the EDL 17 (42%) of motor units survive. This loss of motor units results in muscle weakness and a reduction in force of both muscles. Treatment with MK-801 reduces the effect of nerve injury, so that muscles of treated animals are stronger and weigh more. Cross-sectional area and muscle fibre number in EDL muscles were assessed and found to be dramatically reduced after nerve injury at birth, so that the area was 20% of control, with only 13% of fibres remaining. Moreover the majority of the remaining EDL muscle fibres which are normally fast are converted into slow type I fibres, with 68% of fibres expressing slow myosin compared with 3% in control EDL muscles. In animals treated with MK-801 only 47% of muscle fibres are lost after nerve injury at birth, hence the area of the muscle is greater (51% of control). The change of muscle phenotype induced by nerve injury is prevented and the muscle fibre composition resembles that of normal EDL muscles in that 4% of muscle fibres express slow myosin compared with 3.5% in control EDL muscles. Thus, blocking NMDA receptors with MK-801 shortly after nerve injury at birth reduces the loss of motor units and this is directly reflected in an improved performance of the affected muscles.

Animals↗
PubMed1989

Regulation of expression of avian slow myosin heavy-chain isoforms.

The slow tonic anterior latissimus dorsi (ALD) muscle of the chicken contains two isomyosins, namely SM-1 and SM-2. The proportions of the two isoforms change with age, SM-2 expression increasing at the expense of SM-1. Applying a load on the wing increases the rate and extent of SM-1 replacement. Here we have demonstrated that decreasing the load by removal of the distal portion of the wing in 1-week-old chickens had an effect opposite to that of overloading in that it slowed muscle growth and the rate of SM-1 elimination. Experimental unloading of muscles previously weighted for 1 or 3 weeks slowed the growth rate of muscles, with consequent regression of relative hypertrophy; however, it did not lead to the reexpression of SM-1 myosin. This indicates that the overload-induced changes in myosin expression are not readily reversible. Nerve section produced unexpected results, in that it advanced the normal developmental shift in myosin expression toward predominance of the SM-2 isoform, similar to the effect of muscle overload.

Animals↗
PubMed1989

Muscle fatigue in some neurological disorders.

Fatigue of tibialis anterior (TA) was induced by repetitive electrical stimulation. Using this test, patients with upper motor neuron muscle weakness owing to multiple sclerosis (MS) and injuries to the spinal cord showed greater fatigability of their TA muscles, suggesting that the muscle fiber population changed toward that typical of fatigable motor units. During repetitive stimulation, in addition to the decrement in tension there was an increase in half-relaxation time of tetanic contractions at 40 Hz in both subjects and patients. The increase in half relaxation during repeated activity was greater in patients with MS and spinal cord injury than in healthy subjects, suggesting that the long-term inactivity affected the efficiency of the Ca2+ uptake mechanism of their muscle fibers. Thus long-term inactivity of patients with upper motoneuron dysfunction leads to increased fatigability of their muscles and exaggerates the slowing of muscle relaxation after prolonged exercise.

Electric Stimulation↗
PubMed1989

Long term effect of low frequency chronic electrical stimulation on the fast hind limb muscles of dystrophic mice.

Low frequency chronic electrical stimulation can have a beneficial effect on dystrophic muscles. The present study was undertaken to assess the long term effect of such stimulation on the fast hind limb muscles of dystrophic mice. The relationship between the changes induced by stimulation and the initial condition of the dystrophic muscles, as well as other factors which might contribute to this relationship, were examined. The stimulation induced an increase in the force output of weak dystrophic muscles and a speeding of their time course of contraction and relaxation, as well as an increase in their fatigue resistance. In relatively strong dystrophic muscles, the stimulation induced similar changes in contractile speed and fatigue characteristics, but it led to a slight decrease in force output. Our results suggest that the stimulation promotes the growth and differentiation of the small regenerating fibres known to be present in the diseased muscles and, in addition, induces an increase in the mitochondrial content of the muscle fibres. Our results indicate that these effects are not permanent.

Animals↗
PubMed1988

The distribution of slow myosin in rat muscles after neonatal nerve crush.

Following neonatal nerve injury fast skeletal muscles recover less well than slow ones. This is because many muscle fibers are lost during reinnervation. Since fast muscles normally contain a small population of slow muscle fibers, we have used a monoclonal antibody to slow myosin heavy chains (SMHC) to study their number and pattern of distribution in fast muscles following temporary denervation at 5-6 days of age and subsequent reinnervation. During this time the original distribution of slow fibers changed to one showing irregular grouping, indicating that reinnervation of muscles after neonatal nerve injury is as nonselective as it is after nerve injury in adults. Despite a large reduction in the total number of muscle fibers during reinnervation, the number of slow fibers did not decrease. Thus muscle fiber loss was at the expense of the fast motor units alone.

Animals↗
PubMed1986

The expression of myosin heavy chain isoforms in normal and hypertrophied chicken slow muscle.

Hypertrophy was produced in the anterior latissimus dorsi (ALD) muscle of 5-wk-old chickens by application of a load to the humerus. After 4 wk, hypertrophied ALD muscles were greater than 2.5 times heavier than contralateral control ALD muscles. Two isomyosins are distinguishable in normal ALD muscles by their different electrophoretic mobilities. It is shown here that the faster migrating SM-1 isomyosin decreases in abundance with age and that the application of an overload enhances both the rate and extent of this process. Monoclonal antibodies were selected by an immunotransfer technique that were specific for the heavy chains associated with either SM-1 or SM-2, or cross-reacted with both isoforms. The cellular distribution of the SM-1 and SM-2 isomyosins was analyzed by immunofluorescent technique using these antibodies. Anti-SM-1 and anti-SM-2 antibodies reacted with separate populations of cells, whereas the third antibody reacted with all myocytes in the normal ALD muscle. These data suggest that there is an exclusive cellular distribution of myosin heavy chains associated with SM-1 and SM-2 proteins. Immunofluorescent analysis of hypertrophied muscle showed the anti-SM-2-specific antibody reacting with all myocytes, whereas the anti-SM-1-specific antibody reacted with none. This is consistent with the elimination of the SM-1 isoform in hypertrophied muscles.

Animals↗
PubMed1984

Protease inhibitors reduce the loss of nerve terminals induced by activity and calcium in developing rat soleus muscles in vitro.

The end-plate of a mammalian skeletal muscle fibre is innervated by several axons at the time of birth but by only one axon in the adult. In the rat soleus muscle the transition from polyneuronal to single innervation occurs during the first 2-3 weeks after birth. While it is evident that the loss of the excess nerve terminals depends to some extent on neuromuscular activity, the mechanism involved is not known. In the present experiments neonatal rat soleus muscles were stimulated in vitro in the presence of a variety of combinations of calcium, the cholinesterase inhibitor edrophonium and the proteolytic enzyme inhibitors leupeptin, pepstatin and Ep-475. Electron microscopical examination revealed that stimulation alone had little effect on the morphology of the end-plate region but stimulation in the presence of raised levels of calcium caused severe disruption of the nerve terminals and a marked reduction in the number of intact nerve terminal profiles contacting each end-plate. Contraction measurements showed that, in spite of this, the muscles were not functionally denervated to any large extent. The addition of edrophonium potentiated the morphological alterations but caused no further reduction in the number of profiles. Conversely, the protease inhibitors wholly or partially (in the case of Ep-475) prevented the effects of stimulation and calcium on the nerve terminals. These results are consistent with the idea that neuromuscular activity induces the secretion of proteolytic enzymes into the end-plate region, where they digest the immature nerve terminals. The importance of calcium suggests that the calcium-dependent neutral protease may be involved, and is also consistent with a secretory mechanism. The possibility that the nerve terminals are digested by their own proteases is also discussed.

Acetylcholine↗
PubMed1983

The effect of cross-innervation on the tropomyosin composition of rabbit skeletal muscle.

Soleus, semitendinosus and crureus muscles of the rabbit were found to contain alpha- and beta-tropomyosin subunits and additional forms that have been provisionally designated gamma and delta. Extensor digitorum longus and psoas muscles contained only alpha and beta subunits, the relative proportions of which varied between single fibres of psoas muscle. On cross-innervation of rabbit soleus and extensor digitorum longus muscles, the fraction of the total tropomyosin present as the beta subunit remained constant. The relative proportions of alpha, gamma and delta subunits changed as would be expected from the change in speed that occurred.

Animals↗
PubMedDate not supplied

Duchenne dystrophy viewed as a disturbance of nerve-muscle interactions.

The possibility that some features of Duchenne muscular dystrophy (DMD) can be explained as a disturbance of nerve-muscle interaction during development is discussed. It is argued that the initial disturbance in DMD is due to a slower rate of maturation of skeletal muscle fibers. Normally, the maturation of the motor reflexes is closely followed by maturation of the muscle fibers. The possibility is considered that if muscle fibers mature more slowly than normal, they are not able to withstand the type of activity imposed upon them by the mature motoneuron. This applies mainly to the high frequency activity imposed on the fibers of motor units that are destined to become "fast." Experimental evidence suggesting such a mechanism is presented.

Animals↗