Histochemical, biochemical, and ultrastructural analyses of single human muscle fibers, with special reference to the C-fiber population (original) (raw)
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Journal of Histochemistry & Cytochemistry, 1993
This study characterized histochemically three fast fiber types (IIB, IID, IIA) in skeletal muscles of mouse, rat, and rabbit, with special reference to fiber types IIB and IID. The results are complemented by biochemical analyses of myosin heavy chain composition in these muscles. Fiber type delineation is based on various methods for mATPase staining with pre-incubations and assays under different conditions. In rat and mouse, IIB and IID fibers can be best distinguished according to their different mATPase stabilities towards formaldehyde and alkaline pH. In rabbit, the method of Matoba and Gollnick using acid pre-incubation provided best and most reproducible results. In addition to their different mATPase stabilities, the three fast fiber types differ with regard to their oxidative capacities and cross-sectional fiber areas in the three species. In general, Type IIB fibers are the largest and least oxidative, Type IIA fibers the smallest and most oxidative, and Type IID fibers ...
The Intermediate Muscle Fiber of Rats and Guinea Pigs
Journal of Histochemistry & Cytochemistry, 1969
The soleus, plantaris and gastrocnemius muscles of 60 Sprague-Dawley rats and 70 Hartley guinea pigs were studied histochemically. In the plantaris and gastrocnemius muscles, myosin adenosine triphosphatase activity differentiated intermediate fibers from red and white fibers as determined by malate dehydrogenase and succinate dehydrogenase and reduced nicotinamide adenine dinucleotide diaphorase activities. Contrary to what is commonly reported, red fibers could not be distinguished from white fibers on the basis of myosin adenosine triphosphatase activity as is commonly reported. The intermediate fiber was characterized by minimal menadione-mediated α-glycerophosphate dehydrogenase, phosphorylase and myosin adenosine triphosphatase activity and moderate malate dehydrogenase and succinate dehydrogenase and reduced nicotinamide adenine dinucleotide diaphorase activities. It is suggested that fibers with intermediate oxidative enzyme activity are physiologically slow, white fibers ar...
Human skeletal muscle: transition between fast and slow fibre types
Pflügers Archiv - European Journal of Physiology, 2011
Human skeletal muscles consist of different fibre types: slow fibres (slow twitch or type I) containing the myosin heavy chain isoform (MHC)-I and fast fibres (fast twitch or type II) containing MHC-IIa (type IIA) or MHC-IId (type IID). The following order of decreasing kinetics is known: type IID>type IIA>>type I. This order is especially based on the kinetics of stretch activation, which is the most discriminative property among fibre types. In this study we tested if hybrid fibres containing both MHC-IIa and MHC-I (type C fibres) provide a transition in kinetics between fast (type IIA) and slow fibres (type I). Our data of stretch activation kinetics suggest that type C fibres, with different ratios of MHC-IIa and MHC-I, do not provide a continuous transition. Instead, a specialized group of slow fibres, which we called "transition fibres", seems to provide a transition. Apart of their kinetics of stretch activation, which is most close to that of type IIA, the transition fibres are characterized by large cross-sectional areas and low maximal tensions. The molecular cause for the mechanical properties of the transition fibres is unknown. It is possible that the transition fibres contain an unknown slow MHC isoform, which cannot be separated by biochemical methods. Alternatively, or in addition, isoforms of myofibrillar proteins, other than MHC, and posttranslational modifications of myofibrillar proteins could play a role regarding the characteristics of the transition fibres.
Fibre size and metabolic properties of myosin heavy chain-based fibre types in rat skeletal muscle
Journal of muscle research and cell motility, 1998
Cross-sectional area (CSA), succinate dehydrogenase (SDH), and alpha-glycerophosphate dehydrogenase (GPD) activities were measured in single fibres of adult rat medial gastrocneminus muscle (deep region) using quantitative histochemical procedures. The same fibres were identified in serial sections stained by immunohistochemistry with monoclonal antibodies specific for selected myosin heavy chain (MyHC) isoforms. The values of CSA, SDH and GPD activities formed a continuum, but significant differences in mean values were observed among fibre types of presumed homogeneous MyHC content. Type IIA fibres were the smallest, type IIB fibres were the largest, and type I and IIX fibres were intermediate. Type IIA fibres had the highest SDH activity, followed in rank order by type IIX, type I and type IIB. The average GPD activity was ranked according to fibre type such that IIB > IIX > IIA > I. Hybrid fibres co-expressing two MyHC isoforms generally showed intermediate mean CSA, SD...
Cell Tissue Res, 1996
The relationship between myonuclear number, cellular size, succinate dehydrogenase activity, and myosin type was examined in single fiber segments (n = 54; 9 _+ 3 mm long) mechanically:gdissected from soleus and plantaris muscles of adult rats. One end of each fiber segment was stained for DNA before quantitative photometric analysis of succinate dehydrogenase activity; the other end was double immunolabelled with fast and slow myosin heavy chain monoclonal antibodies. Mean _+ S,D. cytoplasmic volume/myonucleus ratio was higher in fast and slow plantaris fibers (112 _+ 69 vs. 34 _+ 21 x 103 gm 3) than fast and slow soleus fibers (40 _+ 20 vs. 30 + 14 x 103 i, tm3), respectively. Slow fibers always had small volumes/myonucleus, regardless of fiber diameter, succinate dehydrogenase activity, or muscle of origin. In contrast, smaller diameter (< 70 gm) fast soleus and plantaris fibers with high succinate dehydrogenase activity appeared to have low volumes/myonucleus while larger diameter (> 70 gm) fast fibers with low succinate dehydrogenase activity always had large volume/myonucleus. Slow soleus fibers had significantly greater numbers of myonuclei/mm than did either fast soleus or fast plantaris fibers (116 _+ 51 vs. 55 + 22 and 44 _+ 23), respectively. These data suggest that the myonuclear domain is more limited in slow than fast fibers and in the fibers with a high, compared to a low, oxidative metabolic capability.
A histoenzymatic study of rat intrafusal muscle fibres
Histochemistry, 1979
The histochemical activities of myofibrillar adenosine triphosphatase (ATPase), succinic dehydrogenase (SDH) and alpha glycerophosphate dehydrogenase (~-GPD) were studied in intrafusal muscle fibres of rat fast and slow muscles. The ATPase reaction was carried out after the three standard acid preincubations. The cold K2-EDTA preincubated ATPase reaction product was similar to that seen following the regular or alkalipreincubated ATPase reaction, except that the intermediate bag fibres exhibited much higher activity after cold K2-EDTA preincubation. Following either acetic acid solution or cold and room temperature Kz-EDTA-preincubation, followed by the ATPase reaction, chain fibres of the fast muscles vastus lateralis and extensor digitorum longus exhibited a very low amount of reaction product as compared with those of the slow soleus. Veronal acetate and K2-EDTA preincubations (and equally preincubation in acetic acid solution) resulted in acid stable ATPase activity along the entire length of the typical bag fibres but only in the polar regions of the intermediate bag fibres. On the basis of differing ~-GPD reaction, two sub populations of nuclear chain fibres were discovered in one spindle. It is a matter of conjecture, to what extent the histochemical differences of intrafusal fibres from fast and slow muscles reflects functional distinctions in the response to stretch of muscle spindles from fast and slow muscles.
The Journal of physiology, 1987
1. Normal and chronically stimulated peroneus longus muscles of the cat's hind limb were studied with respect to fibre size and staining properties for myofibrillar (myosin) adenosine triphosphatase (ATPase) and succinate dehydrogenase (SDH) activity. The intensity of staining for SDH activity was measured by microphotometry from the central portions of the muscle fibres ('core-SDH staining'). For comparison, histochemical properties were also studied in non-stimulated soleus muscles. 2. On account of the pH sensitivity of their myofibrillar ATPase, about 18% of the fibres in normal peroneus longus muscles were classified as type I, and about half of the remainder as II A and II B respectively. 3. In the normal peroneus longus muscles, the mean diameter of single muscle fibres generally varied between about 25 and 75 micron, whereby the average size of type I less than type II. 4. In the normal peroneus longus muscles the staining intensity for core SDH varied over a wid...
No classical type IIB fibres in dog skeletal muscle
Histochemistry, 1982
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 fibres were combined. Subdivision of type II fibres into two main classes 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 unreactive with anti-IIA myosin, was also oxidative-glycolytic, and only 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 liB fibres could be detected.