PubMed Health⌕ Search

Biomedical subjects

D J Parry

Publications and source records attributed to D J Parry.

At least 37 records · Page 2Linked to original sources

Culturing satellite cells from living single muscle fiber explants.

Conventional methods for isolating myogenic (satellite) cells are inadequate when only small quantities of muscle, the tissue in which satellite cells reside, are available. We have developed a tissue culture system that reliably permits isolation of intact, living, single muscle fibers with associated satellite cells from predominantly fast and slow muscles of rat and mouse; maintenance of the isolated fibers in vitro; dissociation, proliferation, and differentiation of satellite cells from each fiber; and removal of the fiber from culture for analysis.

Animals↗

Satellite cell activity is required for hypertrophy of overloaded adult rat muscle.

Hypertrophy of extensor digitorum longus muscle, overloaded by the removal of the synergist tibialis anterior (TA) muscle, in growing rats is inhibited if endogenous satellite cells are sterilized by exposure to irradiation. However, normal muscle growth is not eliminated, only diminished. To test whether irradiated, overloaded muscle can hypertrophy in the absence of normal growth-related stimuli, experiments were conducted on mature rats. TA muscle ablation caused hypertrophy of EDL muscle, characterized by a significant increase in muscle mass and the size of type IIx and type IIb fibers, and a proportional increase in the number of myonuclei. When ablation was preceded by irradiation, hypertrophy did not occur. The results indicate that satellite cell activation, division, and fusion is necessary for compensatory hypertrophy of fully mature muscle, and may be important to the understanding of the limits of recovery of inherited muscle myopathies treated by myogenic cell implantation.

Adaptation, Physiological↗

Identification, distribution, and myosin subunit composition of type IIX fibers in mouse muscles.

The purpose of the present investigation was to study the distribution and subunit composition of type IIX fibers in mouse muscles. The existence of a population of type IIX fibers in fast-twitch muscles of the mouse was shown by mean of immunohistochemistry and gel electrophoresis. In the hindlimb muscles, tibialis anterior (TA) and extensor digitorum longus (EDL), type IIX fibers account for approximately one third of the total fiber number, with the superficial portion of the TA (TAS) being composed exclusively of type IIB and IIX fibers. A similar proportion of IIX fibers was found in diaphragm (DIA) while in tongue muscles approximately 40% of the fibers were IIX. Single fiber gel electrophoresis revealed a significant number of fibers in TAS that contain both IIB and IIX myosin heavy chain (MyHC). This was confirmed with immunohistochemistry, which revealed the presence of fibers with various degrees of staining intensity. This suggests that there may exist a degree of plasticity which results in the conversion of IIX fibers to IIB fibers and vice versa. Analysis of myosin light chain (MyLC) composition of type IIX fibers revealed that the ratio of MyLC3f to MyLC1f was significantly lower than in type IIB fibers.

Animals↗

Adaptation of rat extensor digitorum longus muscle to gamma irradiation and overload.

The right extensor digitorum longus (EDL) muscle of growing male rats was overloaded by ablation of its synergist tibialis anterior (TA) muscle. Four weeks later, the overloaded muscle was heavier and contained larger type IIA, IIX and IIB fibres than either untreated contralateral muscle or control muscle from an untreated animal. The myonuclear-to-myoplasmic volume ratio was maintained in the overloaded muscle. Overloaded EDL muscle, previously subjected to a dose of irradiation sufficient to sterilise satellite cells, and EDL muscle which had been only irradiated, were significantly lighter and contained significantly smaller fibres than controls, though a significant amount of normal EDL muscle growth did occur following either treatment. The myonuclear-to-myoplasmic volume ratio of the irradiated muscles was smaller than in controls. Overloaded muscle, with or without prior irradiation, possessed a smaller proportion of fibres containing IIB myosin heavy chain (MHC) and a larger proportion of fibres containing IIA and IIX MHC; a significant percentage of these fibres coexpressed either type IIA and IIX MHC or type IIX and IIB MHC. Thus in the absence of satellite cell mitosis, muscles of young rats possess a limited capacity for normal growth but not for compensatory hypertrophy. Adaptations in MHC gene expression to chronic overload are completely independent of satellite cell activity.

Adaptation, Physiological↗

Gamma irradiation prevents compensatory hypertrophy of overloaded mouse extensor digitorum longus muscle.

Mouse extensor digitorum longus (EDL) muscle was subjected to a dose of gamma irradiation that causes reproductive death of satellite cells and/or to chronic compensatory overload, achieved by removal of the distal portion of the tibialis anterior muscle. Four weeks later the mass, fiber type percentage, and fiber size of the EDL muscle were measured. Both the irradiated + overloaded and the irradiated only EDL muscles were significantly lighter and contained significantly smaller fibers than untreated muscle or muscle subjected to chronic overload only. Overload muscle, whether irradiated or not, had a larger percentage of type IIx fibers and a smaller percentage of type IIb fibers than muscle that had not been overloaded. The results confirm that satellite cell proliferation is a prerequisite for muscle hypertrophy induced by synergist incapacitation, but it appears not to be required for the maintenance of, or change in, normal muscle fiber myosin heavy chain phenotype expression.

Animals↗

Tissue distribution of the dystrophin-related gene product and expression in the mdx and dy mouse.

We have previously reported a dystrophin-related locus (DMDL for Duchenne muscular dystrophy-like) on human chromosome 6 that maps close to the dy mutation on mouse chromosome 10. Here we show that this gene is expressed in a wide range of tissues at varying levels. The transcript is particularly abundant in several human fetal tissues, including heart, placenta, and intestine. Studies with antisera raised against a DMDL fusion protein identify a 400,000 Mr protein in all mouse tissues tested, including those of mdx and dy mice. Unlike the dystrophin gene, the DMDL gene transcript is not differentially spliced at the 3' end in either fetal muscle or brain.

Animals↗

The effect of partial denervation of tibialis anterior (TA) muscle on the number and sizes of motorneurons in TA motornucleus of normal and dystrophic (C57BL dy2j/dy2j) mice.

The tibialis anterior (TA) muscle in one leg of normal (C57BL) and dystrophic (dy2j) mice was partially denervated by resection of a part of the lateral popliteal nerve. Two months later the muscle was injected with horseradish peroxidase to permit visualization of the motorneurons that survived. Partial denervation in both C57 and dy2j mice resulted in reduction of the number of motorneurons that supplied the muscle to approximately one-half the normal complement. The surviving motorneurons were found to be significantly larger (about 25%) than their contralateral counterparts. This condition persisted up to 18 months and is not considered to be a transient response to the trauma associated with the partial denervation. When the size of the target tissue was also reduced by extirpation of one-half of TA together with partial denervation, motorneuron size was not found to increase. It is suggested that the increase in size is a response to the metabolic demands placed upon the motorneuron by an increase in the size of the motor unit.

Animals↗

The relationship between post-tetanic potentiation of motor units and myosin isoforms in mouse soleus muscle.

Post-tetanic potentiation was measured in motor units, isolated functionally by ventral root splitting, of soleus and extensor digitorum longus muscles of mouse. All motor units from the extensor digitorum longus had times to peak twitch tension less than 13 ms; there was a linear relationship between time to peak tension and post-tetanic potentiation, with the faster units exhibiting greater potentiation. When soleus motor units were similarly analyzed, it appeared that there may be two distinct populations of units. Those units with times to peak tension less than 13 ms were virtually indistinguishable from those of extensor digitorum longus. On the other hand, the slope of the relationship between post-tetanic potentiation and time to peak tension was significantly lower for soleus units with times to peak tension of 13 ms or more. Approximately three-quarters of the soleus units were of the latter slow type, whereas only one-half of the muscle fibres could be classified as type I by means of immunohistochemistry, suggesting that the myosin heavy chain may not be the major determinant of post-tetanic potentiation. Single, chemically skinned fibres of soleus were analyzed for myosin heavy and light chain components by polyacrylamide gel electrophoresis. All fibres with type I heavy chain contained only the two slow light chains. On the other hand, almost all of the fibres with type IIA myosin heavy chain contained both fast and slow light chains. It is suggested that the discrepancy between the proportions of physiologically "fast" motor units and histochemical type IIA fibres may be the consequence of variable amounts of slow light chain associated with the fast IIA myosin heavy chain.

Animals↗

The effect of reinnervation on the distribution of muscle fibre types in the tibialis anterior muscle of the mouse.

The distribution of fibre types in the tibialis anterior (TA) muscle of adult mice was examined by means of an immunohistochemical approach, using monoclonal antibodies that recognize different myosin heavy chain isoforms. As has been reported previously, the superficial portion of TA contains almost exclusively type IIB fibres and is almost entirely glycolytic in nature. Following section of the lateral popliteal nerve and rotation of the proximal stump to prevent rematching, it was found that the original pattern was virtually restored within 2 months. One possible explanation for this observation is that the activity pattern of peripheral and deep muscle fibres differs and that this aids in specification of muscle fibre type. Alternatively, the muscle fibres of the superficial portion of TA may be inherently resistant to an alteration of their phenotype with regard to expression of myosin heavy chain.

Adenosine Triphosphatases↗

Relative efficacy of slow and fast alpha-motoneurons to reinnervate mouse soleus muscle.

Contractile and histochemical properties of reinnervated motor units in soleus muscles of C57BL/6J mice were examined 1 mo after sectioning the soleus nerve. Fifty-one motor units were isolated by the technique of ventral root splitting. Their sizes ranged from 0.4 to 13.6% of whole muscle tetanic tension (Po) with a mean size of 5.3% Po corresponding to 19 motor units. In control unoperated mice, the range was 2.2-8.6% Po, with a mean size of 4.8% Po corresponding to 22 motor units. Although no clear relationship between unit time to peak tension and size was seen in control units, it appeared that in the reinnervated muscle the large units were also slow contracting, whereas the smaller units were predominantly fast contracting. Adenosinetriphosphatase (ATPase) staining revealed an increase in the proportion of muscle area occupied by type I fibers in reinnervated soleus compared with control soleus. Immunohistochemical staining of reinnervated soleus with monoclonal antibodies against type I and IIa myosin showed the presence of hybrid fibers containing both myosins. It is concluded that during reinnervation most motoneurons reinnervate the soleus muscle of the mouse. The hypothesis that slow motoneurons are more adept at expanding their innervating field than fast motoneurons is also supported by the data.

Adenosine Triphosphatases↗

Number and size of motoneurons in a forelimb motor nucleus of normal and dystrophic (C57BL/6J dy2j/dy2j) mice.

The method of retrograde axonal transport of horseradish peroxidase (HRP) was used to identify the motoneurons that innervate the distal forelimb muscles via the ulnar nerve in normal and dystrophic (C56BL/6J dy2j/dy2j) mice. In both normal and dystrophic mice this motor nucleus was located in spinal segments C6 through T1. No clear division, on the basis of size, into alpha and gamma motoneuron populations was apparent. The motoneurons of dystrophic mice were fewer in number (26.5 vs. 35) but larger in cross-sectional area (780 vs. 674 microns2) than those of age-matched control mice. These results are quantitatively similar to those reported for the hind limb soleus muscle of dystrophic mice and suggest that the motoneuronal changes are a reflection of the dystrophic process rather than the associated spontaneous action potential generation seen in the dystrophic hind limb muscles.

Animals↗

Fatiguability and oxidative capacity of forelimb and hind limb muscles of dystrophic mice.

Fatigue indices and succinic dehydrogenase (SDH) activities were determined in the extensor digitorum longus (EDL) and soleus muscles of the hind limb and the extensor carpi radialis longus of the forelimb in control and dystrophic mice aged 4 to 26 weeks. A good correlation was found between SDH activities and fatigue indices in muscles from normal mice. In the dystrophic (dy2J) mice, however, this correlation was not present. The EDL muscles from 26-week-old dy2J mice showed a much higher resistance to fatigue than age-matched controls but this was not accompanied by a significant change in SDH. The increased fatigue resistance in dy2J EDL appeared between 8 and 12 weeks of age and was temporally correlated with the onset of fused bursts of spontaneous activity in the hind limb muscles. Nevertheless, there was no conclusive evidence for a link among this spontaneous activity, oxidative enzyme capacity, and fatigue resistance.

Animals↗

Succinic dehydrogenase activity of forelimb and hindlimb muscles of the dystrophic mouse.

The activity of succinic dehydrogenase (SDH) was determined in muscles of normal and dystrophic mice. In contradistinction to reports based solely upon histochemical examination, we were unable to observe increased activity in fast-twitch muscles of dystrophic mice. Because dystrophic muscles contain large amounts of connective tissue, two reference bases for expression of enzyme activity were compared. SDH activity was expressed either per micromole of creatine or per milligram of "true muscle fibre weight." The latter was obtained by determining the proportion of the whole muscle occupied by muscle fibres using an image analyzer with photographs of muscle cross section. It appears that the use of creatine content as an index of muscle mass may not be valid for pathological tissue, as the concentration of creatine in some dystrophic muscles differed from that of control muscles. Hindlimb muscles of dystrophic mice exhibit continuous spontaneous activity. To determine the effects of this on oxidative enzyme activity two fast-twitch muscles from the forelimb were also examined. Although they showed histochemical changes comparable to those seen in hindlimb muscles, there was no increase in SDH activity. The only dystrophic muscle examined which showed a change in SDH activity was the soleus in which a decrease was observed.

Animals↗

Motor units in a fast-twitch muscle of normal and dystrophic mice.

Isometric contractions of motor units, isolated functionally by ventral root splitting were recorded from extensor digitorum longus muscles of normal and dystrophic mice of the strain C57BL/6J dy2J/dy2J. Motor unit tetanic tension was significantly lower and both time to peak tension and to half-relaxation of the twitch were significantly prolonged in dystrophic mice relative to age-matched controls. In control mice, motor unit tetanic tension averaged 4.98% of whole muscle tension, corresponding to twenty motor units. Two out of fifteen dystrophic mice exhibited an apparent decrease in the number of motor units, but the data from the remaining thirteen mice indicated no change in relative motor unit size, and hence in the number of motor units. The two mice in which changes were seen were the most severely affected and it is suggested that the apparent reduction in the number of units might be due to some units becoming so small as to be unmeasurable. No evidence was obtained for a population of units with normal characteristics within the dystrophic muscles. There was no clear relationship between tetanic tension and the time to peak tension or to half-relaxation in units from control mice. In dystrophic mice, however, a significant correlation was seen. This possibly reflects two simultaneous effects of the dystrophic process, a loss of tension accompanied by slowing of the twitch.

Animals↗

Isometric contractions of motor units and immunohistochemistry of mouse soleus muscle.

1. Isometric contractions of motor units, isolated functionally by ventral root splitting in vivo, were recorded from mouse soleus muscle. 2. Motor unit tensions varied over a narrow symmetrical range and averaged 4.7% of whole muscle tension, corresponding to twenty-one motor units per muscle. 3. There was considerable variation between muscles in isometric twitch times-to-peak and even greater variation for the motor units. The distribution of motor unit times-to-peak was apparently unimodal and could be fitted by a single normal population. A slightly better fit was, however, obtained with two normal populations, as suggested by the histochemistry. 4. Twitch time-to-peak decreased in proportion to axonal conduction velocity in individual animals. The whole population of motor units could be fitted by a linear relation between time-to-peak and the reciprocal of conduction time in the motor axon. Motor unit tension was also linearly related to the reciprocal of conduction time. 5. Histochemistry showed clear division between Type I and Type IIa fibres. Type I fibres reacted strongly with antibody against slow myosin of cat soleus muscle; Type IIa gave a reaction no stronger than the background. The division was as clear as in the cat or rat.

Animals↗