Stereological analysis of mammalian skeletal muscle (original) (raw)
Related papers
Journal of Ultrastructure Research, 1976
Stereological techniques are used to obtain quantitative information from electron micrographs from 300 fibers of skeletal muscle from the adult guinea pig. Ultrastructural parameters measured include Z line width, volume of mitochondria, volume and surface area of tsystem, terminal cisternae, and sarcoplasmic reticulum. Histograms, scattergrams, and statistical discriminant analysis are used to analyze the data. The whole fiber population can be separated with 90% success rate into the red and white vastus muscles and the soleus muscle, which are physiologically fast and slow, respectively. The soleus fibers have a Z line wider than 1100 A and smaller amounts of t-system and terminal cisternae than the vastus fibers. The mitochondrial volume of the vastus fibers shows a continuous distribution over a large range (0.1-15%), with the white vastus fibers at the lower end and the red vastus fibers at the higher end.
Sizes of components in frog skeletal muscle measured by methods of stereology
Journal of General Physiology, 1975
Stereological techniques of point and intersection counting were used to measure morphological parameters from light and electron micrographs of frog skeletal muscle. Results for sartorius muscle are as follows: myofibrils comprise 83% of fiber volume; their surface to volume ratio is 3.8 mum-1. Mitochondria comprise 1.6% of fiber volume. Transverse tubules comprise 0.32% of fiber volume, and their surface area per volume of fiber is 0.22 mum-1. Terminal cisternae of the sarcoplasmic reticulum comprise 4.1% of fiber volume; their surface area per volume of fiber is 0.54 mum-1. Longitudinal sarcoplasmic reticullum comprises 5.0% of fiber volume, and its surface area per volume of fiber is 1.48 mum-1. Longitudinal bridges between terminal cisternae on either side of a Z disk were observed infrequently; they make up only 0.035% of fiber volume and their surface area per volume of fiber is 0.009 mum-1. T-SR junction occurs over 67% of the surface of transverse tubules and over 27% of th...
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...
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.
Capillary Network in Slow and Fast Muscles and in Oxidative and Glycolytic Muscle Fibres
Image Analysis & Stereology, 2011
The aim of this study was to compare capillary network in slow and fast muscles and also in oxidative and glycolytic muscle fibres. Soleus (SOL) and extensor digitorum longus (EDL) muscles were excised from five female rats. Capillaries and muscle fibres were demonstrated on thick tissue sections by a triple immunofluorescent method. Stacks of perfectly registered optical images were captured by a confocal microscope and further analysed. Applying stereological methods (POINTGRID, FAKIR and SLICER plugin- modules of the Ellipse programme), we estimated the mean length of capillaries, adjacent to individual muscle fibre, per unit fibre length (Lcap/Lfib), per unit surface area of the fibre (Lcap/Sfib) and per unit fibre volume (Lcap/Vfib) in the slow SOL and in predominantly fast EDL muscle, and separately in oxidative and glycolytic fibres of EDL muscle. The length of capillaries per unit fibre length was larger in SOL than in EDL muscle, however, capillary length per unit fibre vol...
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.
Morphofunctional responses to anaemia in rat skeletal muscle
Journal of Anatomy, 2008
Adult male Sprague-Dawley rats were randomly assigned to two groups: control and anaemic. Anaemia was induced by periodical blood withdrawal. Extensor digitorum longus and soleus muscles were excised under pentobarbital sodium total anaesthesia and processed for transmission electron microscopy, histochemical and biochemical analyses. Mitochondrial volume was determined by transmission electron microscopy in three different regions of each muscle fibre: pericapillary, sarcolemmal and sarcoplasmatic. Muscle samples sections were also stained with histochemical methods (SDH and m-ATPase) to reveal the oxidative capacity and shortening velocity of each muscle fibre. Determinations of fibre and capillary densities and fibre type composition were made from micrographs of different fixed fields selected in the equatorial region of each rat muscle. Determination of metabolites (ATP, inorganic phosphate, creatine, creatine phosphate and lactate) was done using established enzymatic methods and spectrophotometric detection. Significant differences in mitochondrial volumes were found between pericapillary, sarcolemmal and sarcoplasmic regions when data from animal groups were tested independently. Moreover, it was verified that anaemic rats had significantly lower values than control animals in all the sampled regions of both muscles. These changes were associated with a significantly higher proportion of fast fibres in anaemic rat soleus muscles (slow oxidative group = 63.8%; fast glycolytic group = 8.2%; fast oxidative glycolytic group = 27.4%) than in the controls (slow oxidative group = 79.0%; fast glycolytic group = 3.9%; fast oxidative glycolytic group = 17.1%). No significant changes were detected in the extensor digitorum longus muscle. A significant increase was found in metabolite concentration in both the extensor digitorum longus and soleus muscles of the anaemic animals as compared to the control group. In conclusion, hypoxaemic hypoxia causes a reduction in mitochondrial volumes of pericapillary, sarcolemmal, and sarcoplasmic regions. However, a common proportional pattern of the zonal distribution of mitochondria was maintained within the fibres. A significant increment was found in the concentration of some metabolites and in the proportion of fast fibres in the more oxidative soleus muscle in contrast to the predominantly anaerobic extensor digitorum longus.