Influence of corticosteroids on myonuclear domain size in the rat diaphragm muscle

2004 ◽  
Vol 97 (5) ◽  
pp. 1715-1722 ◽  
Author(s):  
A. Jeroen Verheul ◽  
Carlos B. Mantilla ◽  
Wen-Zhi Zhan ◽  
Miguel Bernal ◽  
P. N. Richard Dekhuijzen ◽  
...  

Skeletal muscle fibers are multinucleated. Each myonucleus regulates gene products and protein expression in only a restricted portion of the muscle fiber, the myonuclear domain (MND). In the rat diaphragm muscle (DIAm), corticosteroid (CoS) treatment causes atrophy of fibers containing myosin heavy chain (MHC): MHC2X and/or MHC2B. We hypothesized that DIAm fiber MND size is maintained during CoS-induced atrophy. Adult male rats received methylprednisolone for 11 days at 1 (CoS-Low, n = 8) or 8 mg·kg−1·day−1 (CoS-High, n = 8). Age-matched (CTL-AgeM, n = 8), sham-operated (SHAM-AgeM, n = 8), and weight-matched (CTL-WtM, n = 8) animals served as controls. In single DIAm fibers, cross-sectional area (CSA), MND size, and MHC expression were determined. Fiber CSA and MND size were similar in CTL-AgeM and SHAM-AgeM groups. Only fibers containing MHCslow or MHC2A displayed smaller CSA in CTL-WtM than in CTL-AgeM and SHAM-AgeM groups, and MND size was reduced in all fibers. Thus fibers containing MHCslow and MHC2A maintain the number of myonuclei, whereas MHC2X or MHC2B fibers show loss of myonuclei during normal muscle growth. Both CoS groups displayed smaller CSA and MND size than CTL-AgeM and SHAM-AgeM groups. However, compared with CTL-WtM DIAm fibers, only fibers containing MHC2X or MHC2B displayed reduced CSA and MND size after CoS treatment. Thus little, if any, loss of myonuclei was associated with CoS-induced atrophy of MHC2X or MHC2B DIAm fibers. In summary, MND size does not appear to be regulated during CoS-induced DIAm atrophy.

2008 ◽  
Vol 104 (3) ◽  
pp. 787-794 ◽  
Author(s):  
Carlos B. Mantilla ◽  
Rowan V. Sill ◽  
Bharathi Aravamudan ◽  
Wen-Zhi Zhan ◽  
Gary C. Sieck

During early postnatal development in rat diaphragm muscle (Diam), significant fiber growth and transitions in myosin heavy chain (MHC) isoform expression occur. Similar to other skeletal muscles, Diam fibers are multinucleated, and each myonucleus regulates the gene products within a finite volume: the myonuclear domain (MND). We hypothesized that postnatal changes in fiber cross-sectional area (CSA) are associated with increased number of myonuclei so that the MND size is maintained. The Diam was removed at postnatal days 14 (P-14) and 28 (P-28). MHC isoform expression was determined by SDS-PAGE. Fiber CSA, myonuclear number, and MND size were measured using confocal microscopy. By P-14, significant coexpression of MHC isoforms was present with no fiber displaying singular expression of MHCNeo. By P-28, singular expression was predominant. MND size was not different across fiber types at P-14. Significant fiber growth was evident by P-28 at all fiber types (fiber CSA increased by 61, 93, and 147% at fibers expressing MHCSlow, MHC2A, and MHC2X, respectively). The number of myonuclei per unit of fiber length was similar across fibers at P-14, but it was greater at fibers expressing MHC2X at P-28. The total number of myonuclei per fiber also increased between P-14 and P-28 at all fiber types. Accordingly, MND size increased significantly by P-28 at all fiber types, and it became larger at fibers expressing MHC2X compared with fibers expressing MHCSlow or MHC2A. These results suggest that MND size is not maintained during the considerable fiber growth associated with postnatal development of the Diam.


2006 ◽  
Vol 100 (5) ◽  
pp. 1617-1622 ◽  
Author(s):  
Bharathi Aravamudan ◽  
Carlos B. Mantilla ◽  
Wen-Zhi Zhan ◽  
Gary C. Sieck

Denervation (DNV) of rat diaphragm muscle (DIAm) leads to selective atrophy of type IIx and IIb fibers, whereas the cross-sectional area of type I and IIa fibers remains unchanged or slightly hypertrophied. DIAm DNV also increases satellite cell mitotic activity and myonuclear apoptosis. Similar to other skeletal muscles, DIAm fibers are multinucleated, and each myonucleus regulates the gene products in a finite fiber volume, i.e., myonuclear domain (MND). MND size varies across DIAm fiber types in rank order, I < IIa < IIx < IIb [fiber type based on myosin heavy chain isoform expression]. We hypothesized that, after DNV, the total number of myonuclei per fiber does not change and, accordingly, that MND changes proportionately to the change in fiber size regardless of fiber type. Adult rats underwent unilateral (right side) DIAm DNV, and after 2 wk single fibers were dissected. Fiber cross-sectional area, myonuclear number, and MND were measured by confocal microscopy, and these values in DNV DIAm were compared with those obtained in controls. After DNV, type I fibers hypertrophied, type IIa fiber size was unchanged, and type IIx and IIb fibers atrophied compared with control. The total number of myonuclei per fiber was not affected by DNV. Accordingly, after DNV, type I fiber MND increased by 25%, whereas it decreased in type IIx and IIb fibers by 50 and 70%, respectively. These results suggest that MND is not maintained after DNV-induced DIAm fiber hypertrophy or atrophy. These results are interpreted with respect to consequent effects of DNV on myonuclear transcriptional activity and protein turnover.


1982 ◽  
Vol 242 (3) ◽  
pp. C178-C183 ◽  
Author(s):  
C. R. Ashmore

Skeletal muscle growth induced by passive stretch was characterized in the Patigialis muscle of chicks with hereditary muscular dystrophy. When the muscle of 6-wk-old chicks was stretched for 1 wk, the effects on muscle growth and on muscle pathology were variable, but in general few differences between stretched and unstretched muscles were observed. However, when the muscle of 1-wk-old chicks was stretched for 6 wk, the effects on muscle growth and on prevention of pathology were dramatic. Similar to results obtained previously when normal chick muscles were stretched [Holly et al., Am. J. Physiol. 238 (Cell Physiol. 7): C62-C71, 1980; Barnett et al., Am. J. Physiol. 239 (Cell Physiol. 8): C39-C46, 1980], stretched dystrophic muscle increased in weight (200%), cross-sectional area (107%), and fiber cross-sectional area (82%). DNA concentration, which is severalfold higher in unstretched dystrophic muscle compared with unstretched normal muscle, fell to values not different from normal values after being stretched. Nuclei per square millimeter also were the same for stretched dystrophic and stretched normal muscle. Histograms indicated that stretching induced a fiber distribution in dystrophic muscle qualitatively similar to that found in stretched normal muscle. Cytochemical observations revealed a dramatic protective effect of stretch against the progressive pathology of dystrophy. It is concluded that stretch of muscle applied to newly hatched dystrophic chicks is a powerful deterrent of symptoms characteristic of hereditary muscular dystrophy. Stretch imposed after the symptoms of dystrophy are apparent provides little, if any, protection.


1998 ◽  
Vol 84 (4) ◽  
pp. 1407-1412 ◽  
Author(s):  
G. E. McCall ◽  
D. L. Allen ◽  
J. K. Linderman ◽  
R. E. Grindeland ◽  
R. R. Roy ◽  
...  

The purpose of this study was to determine the effects of functional overload (FO) combined with growth hormone/insulin-like growth factor I (GH/IGF-I) administration on myonuclear number and domain size in rat soleus muscle fibers. Adult female rats underwent bilateral ablation of the plantaris and gastrocnemius muscles and, after 7 days of recovery, were injected three times daily for 14 days with GH/IGF-I (1 mg/kg each; FO + GH/IGF-I group) or saline vehicle (FO group). Intact rats receiving saline vehicle served as controls (Con group). Muscle wet weight was 32% greater in the FO than in the Con group: 162 ± 8 vs. 123 ± 16 mg. Muscle weight in the FO + GH/IGF-I group (196 ± 14 mg) was 59 and 21% larger than in the Con and FO groups, respectively. Mean soleus fiber cross-sectional area of the FO + GH/IGF-I group (2,826 ± 445 μm2) was increased compared with the Con (2,044 ± 108 μm2) and FO (2,267 ± 301 μm2) groups. The difference in fiber size between the FO and Con groups was not significant. Mean myonuclear number increased in FO (187 ± 15 myonuclei/mm) and FO + GH/IGF-I (217 ± 23 myonuclei/mm) rats compared with Con (155 ± 12 myonuclei/mm) rats, although the difference between FO and FO + GH/IGF-I animals was not significant. The mean cytoplasmic volume per myonucleus (myonuclear domain) was similar across groups. These results demonstrate that the larger mean muscle weight and fiber cross-sectional area occurred when FO was combined with GH/IGF-I administration and that myonuclear number increased concomitantly with fiber volume. Thus there appears to be some mechanism(s) that maintains the myonuclear domain when a fiber hypertrophies.


1999 ◽  
Vol 87 (2) ◽  
pp. 634-642 ◽  
Author(s):  
Roland R. Roy ◽  
Steven R. Monke ◽  
David L. Allen ◽  
V. Reggie Edgerton

The effects of 10 wk of functional overload (FO), with and without daily treadmill endurance training, on the cross-sectional area, myonuclear number, and myonuclear domain size of mechanically isolated single fiber segments of the adult rat plantaris were determined. The fibers were typed on the basis of high-resolution gel electrophoresis for separation of specific myosin heavy chain (MHC) isoforms and grouped as type I+ (containing some type I MHC with or without any combination of fast MHCs), type IIa+ (containing some type IIa with or without some type IIx and/or IIb but no type I MHC), and type IIx/b (containing only type IIx and/or IIb MHCs). Type I+ fibers had a higher myonuclear number than did both fast types of fibers in the control and FO, but not in the FO and treadmill trained, rats. All fiber types in both FO groups had a significantly larger (36–90%) cross-sectional area and a significantly higher (61–109%) myonuclear number than did control. The average myonuclear domain size of each fiber type was similar among the three groups, except for a smaller domain size in the type IIx/b fibers of the FO compared with control. In general, these data indicate that during hypertrophy the number of myonuclei increase proportionally to the increase in fiber volume. The maintenance of myonuclear domain size near control values suggests that regulatory mechanisms exist that ensure a tight coupling between the quantity of genetic machinery and the protein requirements of a fiber.


1995 ◽  
Vol 79 (5) ◽  
pp. 1629-1639 ◽  
Author(s):  
G. C. Sieck ◽  
W. Z. Zhan ◽  
Y. S. Prakash ◽  
M. J. Daood ◽  
J. F. Watchko

In the rat diaphragm muscle, the histochemical classification of type I, IIa, IIb, or IIx fibers was correlated with myosin heavy chain (MHC) immunoreactivity. Expression of MHC isoforms in single dissected fibers was also assessed electrophoretically. Most fibers (approximately 86%) expressed a single MHC isoform, and when present, coexpression of MHC-2X and MHC-2B isoforms was most prevalent. Type I and IIa fibers were the smallest, type IIb fibers were the largest, and type IIx fibers were intermediate. Succinate dehydrogenase (SDH) and calcium-activated myosin adenosinetriphosphatase (actomyosin ATPase) activities were measured with quantitative histochemical procedures. Type I and IIa fibers had the highest SDH activities, followed in rank order by type IIx and IIb fibers. Type I fibers had the lowest actomyosin ATPase activity, followed in rank order by type IIa, IIx, and IIb fibers. Across all fibers, there was an inverse relationship between fiber SDH activity and cross-sectional area and a positive correlation between fiber actomyosin ATPase activity and cross-sectional area. The SDH and actomyosin ATPase activities of muscle fibers were also inversely correlated. These phenotypic differences in SDH and ATPase activities may be important in determining the contractile and fatigue properties of different fiber types in the rat diaphragm muscle.


1982 ◽  
Vol 47 (3) ◽  
pp. 417-431 ◽  
Author(s):  
K. S. Bedi ◽  
A. R. Birzgalis ◽  
M. Mahon ◽  
J. L. Smart ◽  
A. C. Wareham

1. Male rats were undernourished either during the geslational and suckling periods or for a period of time immediately following weaning. Some rats were killed at the end of the period of undernutrition; others were nutritionally rehabilitated for lengthy periods of time before examination. Two muscles, the extensor digitorum longus (EDL) and soleus (SOL) were studied from each rat. Histochemically-stained transverse sections of these muscles were used to determine total number of fibres, the fibre cross-sectional areas and the relative frequency of the various fibre types.2. All rats killed immediately following undernutrition showed significant deficit sin body-weight, muscle weight and fibre cross-sectional area compared to age-matched controls.3. Animals undernourished during gestation and suckling and then fed normally for 5 months showed persistent and significant deficits in body-weight, muscle weight and total fibre number. There were also significant deficits in mean fibre cross-sectional area of each fibre type except for red fibres in the EDL. No difference in the volume proportion of connective tissue was found.4. Rats undernourished after weaning and then fed ad lib. for approximately 7 months had normal body-and muscle weights. Their muscles showed no significant differences in total fibre number, relative frequency of the various fibre types, fibre size or volume proportion of connective lissue.5. These results indicate that, although the effects on rat skeletal muscle of a period of undernutrition after weaning can be rectified, undernutrition before weaning causes lasting deficits.


1993 ◽  
Vol 74 (5) ◽  
pp. 2450-2455 ◽  
Author(s):  
L. E. Gosselin ◽  
D. A. Martinez ◽  
A. C. Vailas ◽  
G. C. Sieck

The effect of growth on the relative interstitial space [%total cross-sectional area (CSA)] and collagen content of the rat diaphragm muscle was examined at postnatal ages of 0, 7, 14, and 21 days as well as in adult males. The proportion of interstitial space relative to total muscle CSA was determined by computerized image analysis of lectin-stained cross sections of diaphragm muscle. To assess collagen content and extent of collagen maturation (i.e., cross-linking), high-pressure liquid chromatography analysis was used to measure hydroxyproline concentration and the nonreducible collagen cross-link hydroxylysylpyridinoline (HP), respectively. At birth, interstitial space accounted for approximately 47% of total diaphragm muscle CSA. During postnatal growth, the relative contribution of interstitial space decreased such that by adulthood the interstitial space accounted for approximately 18% of total muscle CSA. The change in relative interstitial space occurred without a concomitant change in hydroxyproline concentration. However, the concentration of HP markedly increased with age such that the adult diaphragm contained approximately 17 times more HP than at birth. These results indicate that during development the relative CSA occupied by interstitial space decreases as muscle fiber size increases. However, the reduction in relative interstitial space is not associated with a change in collagen concentration. Thus collagen density in the interstitial space may increase with age. It is possible that the observed changes in relative interstitial space and collagen influence the passive length-force properties of the diaphragm.


1996 ◽  
Vol 8 (3) ◽  
pp. 391 ◽  
Author(s):  
MD Fratacci ◽  
M Levame ◽  
A Rauss ◽  
H Bousbaa ◽  
G Atlan

The changes occurring in the histochemical characteristics of the rat diaphragm during the postnatal period were examined. Fibre-type distribution, fibre oxidative capacity, i.e. succinate-dehydrogenase (SDH) activity, and cross-sectional area were compared in the costal (COS) and crural (CRU) regions, and across their abdominal and thoracic surfaces. The proportions of type I and IIb fibres in both COS and CRU increased with age, while the proportion of type IIa fibres progressively decreased. For COS, fibre distribution was homogeneous over the entire muscle and did not change after 4 weeks. For CRU, it was heterogeneous with a higher proportion of type I fibres on the thoracic surface as from the first week. All fibre types significantly increased in cross-sectional area between 1 and 8 weeks, with no significant differences in COS and CRU. Mean SDH activity did not differ between COS and CRU or across the muscles. Mean SDH activities-were low and identical in all fibre types at birth, and then increased, peaking at the 6th week in type I and IIa fibres. When total muscle fibre oxidative capacity was calculated from an index including fibre-type proportion, cross-sectional area and mean SDH activity, it was significantly higher at 1 than at 8 weeks after birth; this might have functional implications for the newborn.


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