Capillary density and capillary-to-fibre ratio in vastus lateralis muscle of untrained and trained men.

Muscle fibre profile area (Af), volume density (Vv), capillary-to-fibre ratio (CF) and number of capillaries per fibre square millimetre (CD) were determined from needle biopsies of vastus lateralis of twenty-four male volunteers (mean +/- SD: age 25.4+/-5.8 years, height 178.6+/-5.5 cm, body mass 72.1+/-7.7 kg) of different training background. Seven subjects were untrained students (group A), nine were national and sub-national level endurance athletes (group B) with the background of 7.8+/-2.9 years of specialised training, and eight subjects were sprint-power athletes (group C) with 12.8+/-8.7 years of specialised training. Muscle biopsies of vastus lateralis were analysed histochemically for mATPase. Capillaries were visualized and counted using CD31 antibodies against endothelial cells. There were significant differences in the Vv of type I and type II muscle fibres in both trained groups, B (51.8%; 25.6%) and C (50.5%; 26.4%). However, in untrained group A that was treated as a reference group, the difference between Vv of type I and type II fibres was less prominent, nevertheless statistically significant (42.1%; 35.1%). There was also a significant difference in CF: 1.9 in group A and 2.1 in groups B and C. The number of capillaries per mm2 (CD) was 245 (group A), 308 (group B) and 325 (group C). Significant differences (P<0.05) in CF and CD, were found only between group A (1.9; 245) and both groups of trained men, B and C (2.1; 308 and 325). However, endurance athletes (group B), such as long-distance runners, cyclists and cross country skiers, did not differ from the athletes representing short term, high power output sports (group C) such as ice hockey, karate, ski-jumping, volleyball, soccer and modern dance.

Characteristics of myosin profile in human vastus lateralis muscle in relation to training background.

Twenty-four male volunteers (mean +/- SD: age 25.4+/-5.8 years, height 178.6+/-5.5 cm, body mass 72.1+/-7.7 kg) of different training background were investigated and classified into three groups according to their physical activity and sport discipline: untrained students (group A), national and sub-national level endurance athletes (group B, 7.8+/-2.9 years of specialised training) and sprint-power athletes (group C, 12.8+/-8.7 years of specialised training). Muscle biopsies of vastus lateralis were analysed histochemically for mATPase and SDH activities, immunohistochemically for fast and slow myosin, and electrophoretically followed by Western immunoblotting for myosin heavy chain (MyHC) composition. Significant differences (P<0.05) regarding composition of muscle fibre types and myosin heavy chains were found only between groups A (41.7+/-1.6% of MyHCI, 40.8+/-4.0% of MyHCIIA and 17.5+/-4.0% of MyHCIIX) and B (64.3+/-0.8% of MyHCI, 34.0+/-1.4% of MyHCIIA and 1.7+/-1.4% of MyHCIIX) and groups A and C (59.6+/-1.6% of MyHCI, 37.2+/-1.3% of MyHCIIA and 3.2+/-1.3% of MyHCIIX). Unexpectedly, endurance athletes (group B) such as long-distance runners, cyclists and cross country skiers, did not differ from the athletes representing short term, high power output sports (group C) such as ice hockey, karate, ski-jumping, volleyball, soccer and modern dance. Furthermore, the relative amount of the fastest MyHCIIX isoform in vastus lateralis muscle was significantly lower in the athletes from group C than in students (group A). We conclude that the myosin profile in the athletes belonging to group C was unfavourable for their sport disciplines. This could be the reason why those athletes did not reach international level despite of several years of training.

Alterations in kidney morphology in mice with mosaic mutation.

The mosaic (Atp7a(mo-ms)) is an X-linked, lethal mutation in mice. Hemizygous males die at the age of 15 days and they exhibit strong similarities to the brindled and macular mutants. Injection of cupric chloride to mossaic mutants prolongs their life and diminishes the pathological results of mutation. Histochemical analysis of the kidneys from 14-day-old mutant males showed accumulation of copper in the renal cortex of the investigated animals leading to damage of the kidney architecture. A histological profile of the kidneys was defined for four groups of 14-day-old animals: mosaic males ms/-, control males +/-, mosaic males injected with cupric chloride ms/- (Cu), and control males injected with cupric chloride +/- (Cu). Pathological changes were observed in the cortex and in the medulla of the kidneys in both groups of mutants and control males injected with cupric chloride (50 microg of CuCl2 per each individual).

Correlation of mATPase, parvalbumin, and cytochrome c oxidase in rat skeletal muscle fibres.

Muscle fibre typing is conventionally performed on unfixed frozen sections. Within the particular classes of fibre types mATPase correlates well with the type of myosin heavy chain (MHC). However, investigation of other properties, e.g., parvalbumin (PV) level, cannot be determined on unfixed frozen sections. Determination of PV, which correlates with the relaxation time, is performed immunohistochemically on fixed tissue. By that method a comparison of mATPase activity and the amount of PV within the same muscle fibre is difficult. The present method, combining histochemistry of mATPase and cytochrome c oxidase with parvalbumin immunohistochemistry on consecutive sections, allows the distinction of six fibre types: I, IB, IIA, IIB, IIAB, IIX, and a C fibre population without division into IC and IIC fibres. The advantages and disadvantages of the present method are discussed.

Parvalbumin in skeletal muscles of teleost (Tinca tinca L. and Misgurnus fossilis L.). Histochemical and immunohistochemical study.

Musculatures of two fish species belonging to the suborder Cyprinoidei were examined histochemically and immunohistochemically for demonstration of mATP-ase activity and parvalbumin content, respectively. The tench lateral musculature shows a typical "acid reversal" picture of mATP-ase activity--red fibres are alkali labile, acid stable and white and white fibres are alkali stable, acid labile. In the pond loach a superficial layer of musculature is composed of alkali--and acid stable red fibres. This alkaline stability of the red fibres caused the classical "acid reversal" picture impossible to obtain. For immunohistochemical reaction a monoclonal antibody specific for parvalbumin II component of the Carp muscles (Pv Carp II) was used. Immunohistochemical results gave evidence that lateral musculatures of the two species examined possessed parvalbumin II component entirely in the fast-contracting fibres--intermediate and white. This results correlate with mATP-ase activity in the tench musculature and did not correlate in the pond loach with respect to mATP-ase alkali stable red muscle fibres in the fish. We conclude that fish muscles mATP-ase activity not always correlates with parvalbumin content in the opposite to mammalian muscles where such correlation seems to be obvious.

Eye muscles of the pike (Esox lucius) fiber types and their actomyosin ATPase and SDH activity.

The eye muscles of the pike are organized in two distinct populations of fibers arranged in semicircular layers. The distal layer, consisting of small diameter muscle fibers, is mainly made up of type I and tonic (slow) fibers. The central and proximal (facing the eyeball) parts of the eye muscles are occupied by IIB and IIC (fast) fibers. As a main criterion for identification of muscle fiber types, the reaction for actomyosin ATPase activity has been used: metabolic characterization of these fibers has been carried out on the basis of SDH activity.

Histochemical study of the forelimb muscles in Rana temporaria L.

Fibre types in the 7 forelimb muscles of Rana temporaria were characterized according to actomyosin ATP-ase activity and 5 distinct fibre types were distinguished. Besides the twitch fibres of type I, IIA, IIB, and IIC, also the tonic fibres were found in the examined muscles. The metabolic profiles of these fibre types was identified on the basis of SDH activity. The scores of SDH activity of different fibre types overlapped and correlation between fibre types defined by ATP-ase activity and their metabolic profiles was not found. All investigated muscles showed the heterogenous distribution of fibre types. Caput superius m. extensoris carpi radialis and caput mediale m. anconei were composed of 4 and 3 fibre types, respectively, in the opposite to the other ones which contained all 5 types of fibres.

Histochemical characterization of the muscle fiber types of the Teleost (Esox lucius L.).

On the base of the actomyosin ATP-ase activity and succinate dehydrogenase (SDH) activity, 4 types of muscle fibers were described in the pike (Esox lucius L.) body musculature. Besides the typical red, intermediate, and white fibers, probably other fibers discerned were of tonic type. Another fibers found were the intermediate fibers of a characteristic small diameter and localization beyond the intermediate fiber zone. This particular construction of the body musculature is characteristic for darting behaviour fish.

Muscle fibre composition of the eye muscles of the carp (Cyprinus carpio L.) defined on the base of histochemical observations.

A histochemical study has been carried out on the eye muscles of the carp. On the base of the ATP-ase and SDH activity and with regard to the localization and diameter of muscle fibres six types of muscle fibres situated in the defined zones can be distinguished. The type 1 and 2 fibres display a moderately high ATP-ase activity at pH 9.4 and rather low activity after alkaline and acid preincubation. Type 1 shows a high SDH activity in contrast to type 2 with a low SDH activity. The other four types of fibres have the high ATP-ase activity at pH 9.4. Type 3 contains fibres with a moderately high ATP-ase activity after alkaline preincubation with a rather low activity after acid preincubation and with a low SDH activity. The fibres of type 4 characterized by the high ATP-ase activity after alkaline and acid preincubation and by the high SDH activity. The fibres of type 5 display high ATP-ase activity after alkaline and acid preincubation and the low SDH activity. They are situated in the white and intermediate fibre zones. The fibres of type 6 are comparable to the fibres of type 5, however they differ diameter and localization, i.e. they are situated in small diameter fibre zone. Using the electron microscope four types of fibres (A, B, C, and D) are found. They vary in the localization of T system, the organization of Z-line and in M-line appearance. In the type B of muscle fibres two different localizations of T system have been discerned.