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G Zacharová

Publications and source records attributed to G Zacharová.

11 recordsLinked to original sources

The effect of a unilateral muscle transplantation on the muscle fiber type and the MyHC isoform content in unoperated hind limb slow and fast muscles of the inbred Lewis rats.

To reveal the effect of foreign innervation and altered thyroid status on fiber type composition and the myosin heavy chain (MyHC) isoform expression in the rat slow soleus (SOL) and fast extensor digitorum longus (EDL) muscles, a method of heterochronous isotransplantation was developed. In this experimental procedure, the SOL or EDL muscles of young inbred Lewis rats are grafted either into the host EDL or SOL muscles of adult rats of the same strain with normal or experimentally altered thyroid status. To estimate the extent of fiber type transitions in the transplanted muscles, the SOL and EDL muscle from the unoperated leg and unoperated muscles from the operated leg could be legitimately used as controls, but only when the experimental procedure itself does not affect these muscles. To verify this assumption, we have compared the fiber type composition and the MyHC isoform content of unoperated contralateral SOL and EDL muscles and ipsilateral unoperated SOL muscle of experimental rats after unilateral isotransplantation into the host EDL muscle with corresponding muscles of the naive rats of the same age and strain. We provide compelling evidence that the unilateral heterochronous isotransplantation has no significant effect on the fiber type composition and the MyHC isoform content of unoperated muscles of experimental animals. Hence, these muscles can be used as controls in our grafting experiments.

Adenosine Triphosphatases↗

Influence of chronically altered thyroid status on the activity of liver mitochondrial glycerol-3-phosphate dehydrogenase in female inbred lewis rats.

The activity of liver mitochondrial flavoprotein-dependent glycerol-3-phosphate dehydrogenase (GPDH) is considered a reliable marker of thyroid status in acute and short-lasting experiments. The aim of this study was to ascertain whether GPDH activity could also be used as an index of thyroid status during chronic experiments over several months. We therefore analyzed GPDH activity in liver mitochondria of female inbred Lewis rats with thyroid status altered for 2 to 12 months. Hyperthyroid state was maintained by triiodothyronine (T (3)) or thyroxine (T (4)) administration, while methimazole was employed for inducing hypothyroidism. We found a seven- and three-fold increase of GPDH activity in female rats after T (3) or T (4) administration, respectively, compared to euthyroid females (8.9 +/- 2.3 nmol/min/mg protein), whereas administration of methimazole reduced the enzyme activity almost to one-third of the euthyroid values. These changes were not significantly influenced by the duration of hyperthyroid or hypothyroid treatment. We conclude that the level of the rat liver GPDH activity could serve as a useful marker for evaluation of hyperthyroid and hypothyroid status in chronic long-lasting experiments on female inbred Lewis rats.

Animals↗

Influence of thyroid status on the differentiation of slow and fast muscle phenotypes.

Muscle phenotype is determined by combined effects of intrinsic genetic and extrinsic factors like innervation, hormonal levels and mechanical factors or muscle activity. We have been studying the effect of altered thyroid hormone levels on the expression of myosin heavy chain (MyHC) isoforms in control and regenerating soleus and extensor digitorum longus muscles of euthyroid, hypothyroid or hyperthyroid female inbred Lewis rats. The fiber type composition has been determined according to the mATPase activity and immunocytochemical staining of MyHC isoforms, the content of MyHC isoforms has been determined by SDS-PAGE, the mRNA levels have been measured by RT-PCR and the ultrastructural transformation has been analyzed by electron-microscopy. Our results indicate that although the innervation plays a decisive role in the determination of muscle phenotype, levels of thyroid hormones contribute to the extent of muscle phenotype transformation.

Animals↗

Body, heart, thyroid gland and skeletal muscle weight changes in rats with altered thyroid status.

In the present paper we describe changes of anatomical parameters in inbred Lewis strain rats, namely their body weight, body weight gain per week, absolute and relative heart, thyroid gland and skeletal muscle weights, that are assumed to reflect experimentally altered thyroid status. The hyperthyroid state was induced by DL-thyroxine or Na 3,3',5-triiodo-L-thyronine, while methimazole was employed for inducing hypothyroidism. We have found that when compared to euthyroid rats, hypothyroidism resulted in a significantly lower body weight gain, absolute and relative heart weight and, in contrast, in a significant increase of absolute and relative thyroid gland weight. On the other hand, hyperthyroidism led to a significant increase of absolute and relative heart weight and to a significant reduction of absolute and relative thyroid gland weight. However, the body mass was not significantly altered in hyperthyroidism as compared with euthyroid rats. We conclude that our protocol leads to chronic hyper- or hypothyroidism as demonstrated by body, heart and thyroid gland weight changes. These anatomical data can thus be utilized as supplemental criteria for the assessment of the thyroid state of experimental rats.

Animals↗

Expression of myosin heavy chain (MyHC) isoforms in rat intrafusal muscle fibres after neonatal deefferentation and subsequent denervation.

The analysis of developing intrafusal fibres is not feasible in the absence of primary sensory axons, as neonatal denervation leads to the disintegration of muscle spindles. On the other hand, neonatal deefferentation does not arrest their differentiation and, moreover, it leads to the neomyogenesis of supernumerary intrafusal profiles. If the sciatic nerve was sectioned in 4-week-old rats deefferented at the birth, muscle spindles survived, the neomyogenesis proceeded and the denervated intrafusal fibres expressed the spindle specific slow tonic (STO) MyHC. The expression of MyHC pattern in individual fibres and the differentiation of the fibre type characteristics were, however, less obvious compared to the control or deefferented spindles. The newly formed intrafusal profiles (which differentiated from satellite cells in the absence of innervation) expressed the STO MyHC particularly when they developed in a spatial relation to nuclear bag fibres.

Animals↗

Stereological evaluation of the soleus muscle isografted into fast extensor digitorum longus (EDL) muscle in rats with different thyroid status.

The 2-D stereology can be used advantageously in the case of muscle cross sections stained by routine histochemical and immunocytochemical methods, such as mATPase reaction, when the quality of the image is often not sufficient for using image analysis techniques without considerable individual intervention. Other advantages of stereological methods in muscle morphometry are that measurements are made directly on specimens under the microscope and in their simplest arrangement they do not require sophisticated and expensive technical equipment. Furthermore, unbiased results are obtained, no segmentation and edge effect problems arise and the quantity of work invested in stereological estimation is reasonable. Therefore, we have used the stereological methods as our standard technique for assessment of fibre type composition in regenerated soleus muscles grafted from 21- to 28-day-old rats into fast EDL muscles of adult inbred recipients with different plasma levels of thyroid hormones.

Animals↗

Reinnervation of cat pacinian corpuscles after nerve crush.

The reinnervation pattern of crural pacinian corpuscles was examined by light and electron microscopy in eight adult cats of both sexes 3-18 months after sciatic nerve crush. Normal pacinian corpuscles are each supplied with a single myelinated axon and a single cylindrical axon terminal which may branch in the distal part of the inner core. Reinnervation of these vibroreceptors was very satisfactory after sciatic nerve crush: in a sample of 68 corpuscles examined 3-18 months after the operation, 92.6% were found reinnervated, while only 7.4% remained denervated. At the nerve entry, 84.2% of the reinnervated corpuscles were supplied with a single myelinated axon, while 15.8% received two myelinated axons; some of the axons branched before and after entering the inner core. Near the mid-level of the inner core, 60.3% of 63 reinnervated corpuscles were innervated with a single axon terminal, 22.2% were bilateral, while 17.5% had three or more terminals. Regenerated axon terminals induced the formation of thin lamellar layers in the axial region of the original core and, exceptionally, also at the outer aspect of the original core. In monoterminal corpuscles, the shape and ultrastructure of regenerated endings resembled those of normal controls, whereas in multiterminal corpuscles their shape and profiles were variable. In contrast to previous reports, reinnervated corpuscles did not ultimately become monoterminal. On the contrary, the mean number of 1.3 terminals found in reinnervated crural corpuscles at 3-5 months increased to 1.9 terminals per corpuscle 6-18 months after axotomy.

Animals↗

Nociceptive atrophy of the rat soleus muscle induced by bone fracture: a morphometric study.

Reflex atrophy of the soleus muscle induced by ipsilateral metatarsal bone fracture in Sagatal-anesthetized adult male rats was studied by using two-dimensional stereological methods 7 days after the operation. When compared with contralateral solei, the wet weight of the experimental soleus muscles was decreased by approximately 24% and the area of the entire muscle section by approximately 29%. In atrophied solei, the number of type 1 fibers was lower by approximately 8%, resulting in lower total number of fibers (by approximately 6%). This indicates that slow motor units might be more sensitive to nociceptive stimulation. However, with respect to the fiber area, the reflex atrophy induced by metatarsal bone fracture in the rat soleus muscle resembles simple atrophy after 7 days, as the mean muscle fiber area was decreased by approximately 26% with no significant difference between atrophy in type 1 and type 2a fibers (by 27.3 and 23.0%, respectively).

Animals↗

Stereological methods based on point counting and unbiased counting frames for two-dimensional measurements in muscles: comparison with manual and image analysis methods.

Stereological methods, using the principle of point counting and unbiased counting frames, for the estimation of muscle area, total fibre area, number of muscle fibres and mean fibre area are described in detail. Their practical application is demonstrated on cross-sections of the rat soleus muscles. It is shown that the efficiency of these methods is high and their results are comparable with those achieved by the conventional manual and image analysis methods. The main advantages of two-dimensional stereological methods in muscle morphometry are pointed out: measurements are made directly on specimens under the microscope and in the simplest implementation do not require sophisticated and expensive technical equipment. Furthermore, unbiased results are obtained, no segmentation and edge effect problems arise and the quantity of work invested in stereological estimation is reasonable. Based upon the study of the efficiency of used stereological methods, a suitable test system for muscle morphometry is proposed.

Animals↗