Expression of developmentally regulated muscle proteins in rhabdomyosarcomas.

Human skeletal muscle differentiation and maturation follows a precise sequence of events. To investigate whether and to what extent rhabdomyosarcoma (RMS) cells follow a comparable sequence, 29 fresh frozen specimens of RMS (14 primary and 15 relapses) were immunostained with antibodies directed against developmentally regulated myosin heavy chains (MHC), ie, fetal, fast, and slow MHC, in addition to desmin and vimentin. Four distinct patterns of expression were observed: I) RMS cells expressing exclusively vimentin and desmin (n = 7), II) in addition to expression of vimentin and desmin, a minority of neoplastic cells were immunoreactive with fetal MHC (n = 6), III) in addition to pattern II, fast MHC was expressed (n = 7), and IV) RMS cells simultaneously expressing vimentin, desmin, fetal, fast, and slow MHC (n = 9).

Clones of human satellite cells can express in vitro both fast and slow myosin heavy chains.

In order to test the diversification among satellite cells in man, satellite cells were isolated from human quadriceps and masseter muscles. The growth kinetics and morphological features of these cells were determined in vitro and the expression of the different myosin heavy (embryonic, fetal, fast, and slow) and light chain isoforms was analyzed. In all satellite cell cultures, only the four fast-type light chains (MLC1emb, MLC1F, MLC2F, and MLC3F) were synthesized and no slow myosin light chains were ever detected.

Expression of myosin isoforms and of desmin, vimentin and titin in Tunisian Duchenne-like autosomal recessive muscular dystrophy.

Morphological, morphometrical, histoenzymological, immunocytochemical and biochemical analysis were performed on muscle biopsies taken from patients suffering from tunisian autosomal recessive Duchenne-like muscular dystrophy (TDLMD) selected both by Duchenne-like clinical criteria and by the presence of normal dystrophin. Data were compared to that obtained from DMD biopsies characterized by the absence of dystrophin. The distribution of myosin heavy chain isoforms, desmin, vimentin and titin were determined in type I and type II muscle fibers.

Biochemical and immunocytochemical analysis in chronic proximal spinal muscular atrophy.

Immunocytochemical and biochemical analyses were carried out on patients affected by chronic SMA. Three groups of patients were identified. In group I, the muscle presented a fascicular atrophy; a high percentage of atrophic type II fibers; and fibers expressing fast, slow, embryonic, and fetal myosin isoforms.

Immunohistochemical characterization of human masseter muscle spindles.

An enzyme- and immunohistochemical study has been performed on human masseter muscle spindles. Antibodies selective for different myosin heavy chain (MHC) isoforms and M-band proteins (M-protein, myomesin, and MM-CK) were used. The expression of these proteins was determined in the different intrafusal fiber types.

[Main steps in the skeletal myogenesis in man].

During the metabolic differentiation of developing human skeletal muscle fibers, two different generations of fibers can be characterised by their profiles of protein expression (in particular of the myosin heavy chains). This differentiation is regulated by both genetic and epigenetic factors (motor innervation, levels of thyroid hormone). The anomalies in different muscle pathologies can be better understood by comparison with the modifications occurring during normal skeletal muscle development.

[Development of the diaphragmatic muscle].

Muscle biopsies were taken at the oesophageal opening and in the costal part of the diaphragm from 15 foetuses (aged between 11 and 40 weeks) and from four children aged 1 week, 5 months, 3 and 16 years. Subjects with multiple congenital abnormalities or neuromuscular diseases were not included in this study. Enzyme histochemistry (myosin ATPases) and immunohistochemistry using antibodies directed against vimentin, desmin, titin and different isoforms of the myosin heavy chains (MHCs) were carried out on serial frozen sections.

Plasticity of human satellite cells.

Satellite cells were isolated from human quadriceps and masseter muscles and the phenotype of these cells examined in vitro. The expression of the different isoforms of the myosin heavy chains (embryonic, fetal, fast and slow) and light chain isoforms was used to assay myotube diversification. We found that fused cultures of human satellite cells express adult fast and slow MHCs in addition to the embryonic and fetal isoforms.

Modification of the dystrophic phenotype after transient neonatal denervation: role of MHC isoforms.

While it recently has been demonstrated that it is possible to modify the phenotypic expression of murine dystrophy (dy/dy) (i.e., prevent myofiber loss) by subjecting the extensor digitorum longus (EDL) muscle of 14-day-old dy/dy mice to transient neonatal denervation (Moschella and Ontell, 1987), the mechanism responsible for this phenomenon has not been determined.

[The characteristic phenotype of the masseter muscle fibers is established after birth].

Muscle biopsies were taken from the superficial portion of the M. masseter in 10 foetuses (aged between 12 and 38 weeks), in a child of 18 months and in an adult without any neuromuscular disease. Serial frozen sections were processed for immunocytochemistry using antibodies specific for the embryonic, foetal, slow and fast myosin heavy chains (MHCs).

Expression of alpha-cardiac myosin heavy chain in mammalian skeletal muscle.

We have investigated the reactivity of different human, rat and cat muscles to a monoclonal antibody directed against human alpha-cardiac myosin heavy chain. We have found that special fiber subpopulations of human masseter and extraocular muscles, as well as the bag fibers of human, rat and cat muscle spindles, were reactive to this antibody, indicating that these fibers expressed alpha-cardiac myosin heavy chain or a closely related isoform. This isomyosin was present in the spindle bag fibers at early fetal stages, whereas its expression in masseter and extraocular muscle fibers was not detected during the first 22 weeks of gestation.

Evolution of muscle specific proteins in Werdnig-Hoffman's disease.

The pattern of expression of desmin, vimentin, titin and different myosin isoforms expressed in atrophic and hypertrophic type I and type II muscle fibers was investigated in 7 biopsies from patients of various ages all diagnosed as suffering from Werdnig-Hoffman's disease. The results revealed that there was a progressive atrophy affecting both type I and type II muscle fibers. The proportion of atrophic type II fibers increased with age.

Fetal development of the urethral sphincter.

A morphological study was carried out on 25 human embryos (3-60 mm CR) and 20 human fetuses (15-40 weeks). The anatomical analysis was completed by immunocytochemical studies of different markers of muscle differentiation (vimentin, desmin, titin and isoforms of the myosin heavy chains). A mesenchymal condensation forms around the urethra after the division of the cloaca (E.

Developmental expression of troponin I isoforms in fetal human heart.

We have used antibodies specific for troponin I proteins to examine human cardiac development and have detected a transiently expressed developmental isoform. This isoform is distinct from adult cardiac troponin I (TnIc) but is indistinguishable, on the basis of electrophoretic mobility and antibody reactivity, from the isoform found in slow skeletal muscle (TnIs). Furthermore, we show that mRNA for TnIs is present in fetal, but not adult, heart.

Transcription of the embryonic myosin light chain gene is restricted to type II muscle fibers in human adult masseter.

We have previously demonstrated that the embryonic myosin light chain (MLC1emb) isoform whose expression is restricted to the early fetal stages in most mammalian skeletal muscles, persists throughout development in human masseter muscle. In order to go further in this study, we have compared the developmental profile of MLC1emb gene transcription in human masseter and quadriceps muscles using both Northern blotting and in situ hybridization techniques. Interestingly, whereas expression of this gene was observed in all fibers during fetal stages in both muscles, transcription in adult masseter was found to be restricted to type II fibers.

[Quantification of the diameter of muscular fibres in the course of the development of the quadriceps].

Muscle biopsies were taken from the medium portion of the vastus lateralis of a hundred fetuses (aged between 10 and 40 weeks), children, adolescents and adults without any neuro-muscular disease. Diameters of the muscle fibers were measured with a Leitz ASM 68K on frozen sections which had been stained for myofibrillar ATPase. The results have been expressed as average diameters and histograms.

Mechanogenetic regulation of transcription.

In many biological systems mechanical forces regulate gene expression: in bacteria changes in turgor pressure cause a deformation of the membrane and induce the expression of osmoregulatory genes; in plants gravity regulates cell growth ('geotropism'); in mammals stretching a muscle induces hypertrophy which is accompanied by qualitative changes in protein synthesis. Consequently, the term 'mechanogenetic control' seems to be a suitable common name for all these processes. The mechanism by which mechanical factors modulate transcriptional activity is still unknown.

Effects of perinatal undernutrition on elimination of immature myosin isoforms in the rat diaphragm.

The effect of perinatal undernutrition on the postnatal elimination of immature myosin isoforms in rat diaphragm muscle was examined using electrophoretic and immunocytochemical techniques. Electrophoresis of native myosin showed that neonatal bands were present in diaphragm muscles of both control and undernourished rats on day 4. By day 21, the neonatal bands were diminished in the control diaphragm compared with the diaphragm of the undernourished rats.

Immunocytochemical characterisation of two generations of fibers during the development of the human quadriceps muscle.

We have carried out a comprehensive study of the formation of muscle fibers in the human quadriceps in a large series of well dated human foetuses and children. Our results demonstrate that a first generation of muscle fibers forms between 8-10 weeks. These fibers all express slow twitch myosin heavy chain (MHC) in addition to embryonic and foetal MHCs, vimentin and desmin.

Modification in the expression and localization of contractile and cytoskeletal proteins in Schwartz-Jampel syndrome.

Muscle biopsies taken from 4 patients with clinical diagnosis of Schwartz-Jampel syndrome were analyzed by enzyme-histochemical immunocytochemical and biochemical techniques. In situ distribution of the different myosin heavy chain (MHC) isoforms together with that of the cytoskeletal proteins vimentin, desmin and titin was determined in type I, type IIA, type IIB and type IIC fibers. The same muscle biopsies were analyzed for their content in myosin light chains (MLC) by two-dimensional gel electrophoresis and native myosin isoforms by pyrophosphate gel electrophoresis.

Biphasic expression of slow myosin light chains and slow tropomyosin isoforms during the development of the human quadriceps muscle.

Using a two-dimensional electrophoresis technique coupled with sensitive silver staining, we have investigated the chronology of appearance of the myosin light chain and tropomyosin isoforms during early stages of human quadriceps development. Our results show that slow myosin light chains and the slow tropomyosin isoform are not detected at 6 weeks of gestation. These isoforms transiently appear between 12.

Emergence of the mature myosin phenotype in the rat diaphragm muscle.

Immunohistochemical analysis of myosin heavy chain (MHC) isoform expression in perinatal and adult rat diaphragm muscles was performed with antibodies which permitted the identification of all known MHC isoforms found in typical rat muscles. Isoform switching, leading to the emergence of the adult phenotype, was more complex than had been previously described. As many as four isoforms could be coexpressed in a single myofiber.

Distinct contractile protein profile in congenital myotonic dystrophy and X-linked myotubular myopathy.

The contractile proteins present in muscle biopsies taken from infants suffering either from congenital myotonic dystrophy or X-linked myotubular myopathy were compared using biochemical and immunocytochemical techniques. Two-dimensional gel analysis has revealed that in all cases of X-linked myotubular myopathy the pattern of expression of myosin light chains, tropomyosin and troponin was roughly similar to that of normal age matched control muscle. However, biopsies from infants affected by congenital myotonic dystrophy demonstrated a predominance of most fast contractile protein isoforms.

Molecular cloning of human cardiac troponin I using polymerase chain reaction.

We have used the polymerase chain reaction (PCR) to synthesise a cDNA encoding part of human cardiac troponin I. Amplification was achieved using fully degenerate sets of oligonucleotides corresponding to conserved regions of amino acid sequence identified in other troponin I isoforms. The cloned PCR fragment was subsequently used to isolate full-length cDNAs from a cardiac cDNA library.

Expression of myosin isoforms during notexin-induced regeneration of rat soleus muscles.

Myosin isozymes and their fiber distribution were studied during regeneration of the soleus muscle of young adult (4-6 week old) rats. Muscle degeneration and regeneration were induced by a single subcutaneous injection of a snake toxin, notexin. If reinnervation of the regenerating muscle was allowed to occur (functional innervation nearly complete by 7 days), then fiber diameters continued to increase and by 28 days after toxin treatment they attained the same values as fibers in the contralateral soleus.