[Postnatal development of the masticatory muscles of the Wistar rat (Rattus norvegicus Berkenhout). A histochemical study]. / Postnatale Entwicklung der Kaumuskulatur der Wistarratte (Rattus norvegicus BERKENHOUT). Eine histochemische Studie.

Muscle biopsies of the mandible adductors of the Wistar rat (Rattus norvegicus Berkenhout) have been analyzed enzyme-histochemically for the investigation of the postnatal development (42.-126. d post partum) of their muscle fibers with special regards to the fiber types. The following methods have been used in this investigation: myofibrillar adenosine triphosphatase (ATP-ase) with different pre-incubations, Sudan black B (triglycerides), periodic acid-Schiff reagent (PAS) (glycogen), Gomori modified Trichrome staining. Quantitative analysis of muscle fibre-type composition and muscle fibre size was done from different regions of muscle sections. Estimation of the fiber size was carried out by measuring the minimum diameter of each type of muscle fibre.

Immunocytochemical analysis of the perinatal development of cat masseter muscle using anti-myosin antibodies.

The developmental changes in myosin gene expression in the masseter muscle of embryonic and juvenile kittens were examined immunocytochemically using anti-myosin heavy chain antibodies of various specificities. In the mature cat, this muscle contains only two phenotypes, the majority of fibres are superfast, the rest being slow fibres. In foetal tissues, the histological appearance of bundles of myotubes, comprising a large central myotube surrounded by a rosette of smaller myotubes, strongly suggest the existence in the jaw muscle of primary and secondary fibres during development. Immunocytochemical data are consistent with the hypothesis that there are four types of fibre; two types of primary fibre as well as two types of secondary fibre. (1) Slow primaries stain strongly with an anti-slow myosin antibody throughout the period under study. These fibres transiently express embryonic but not foetal myosin. (2) Superfast primaries stain for embryonic/foetal and slow myosins in the perinatal period but progressively replace these myosins with superfast myosin during postnatal development. (3) Superfast secondaries initially express embryonic/foetal myosins, but later, beginning around the time of birth progressively replace these myosins with superfast myosin. These fibres do not express slow myosin. (4) Slow secondaries, which initially also express embryonic/foetal myosins, but which postnatally express slow or slow and superfast myosins and express only slow myosin in the adult. These four types of fibres are homologous to the four isotypes of limb muscle fibres and may be derived from distinct lineages of myoblasts.

Histochemical characteristics of masseter and temporalis muscles after 5 weeks of maxillomandibular fixation--an investigation in Macaca mulatta.

This study describes the histochemical characteristics and cross-sectional areas of the superficial masseter and temporalis muscles in juvenile rhesus monkeys after 5 weeks of maxillomandibular fixation. Four juvenile male Macaca mulatta underwent mandibular surgery and 5 weeks of maxillomandibular fixation as part of a study of temporomandibular joint (TMJ) adaptations after condylar replacement. Immediately before the time the animals were killed (5 weeks postsurgically), biopsies of the masseter and temporalis muscles were obtained and submitted to histochemical analysis and calculation of muscle-fiber areas. The data were compared to histochemistry from 12 juvenile control Macaca mulatta. Significant decreases in mean cross-sectional area were exhibited in both type I (p less than 0.05) and type II (p less than 0.01) fibers in all muscles when compared to controls (n = 12). The ratio of type I to type II fibers, however, remained constant during maxillomandibular fixation in masseter and temporalis muscle samples, indicating no change in relative types of fibers. We conclude from this experimental investigation that (1) significant atrophy occurs in the temporalis and masseter muscles after 5 weeks of maxillomandibular fixation, and (2) this atrophy occurs in both type I and type II fibers, indicating that overall recruitment of the muscle (and not just of one fiber type of motor unit) was affected during fixation.

Changes in distribution of some crucial elements in masseter myofibers following surgically induced trauma.

Experimentally induced surgical trauma of the masseter muscle in rats led to changes in the distribution of some crucial intracellular elements as determined by microprobe analysis. Sulfur, phosphorus, and potassium values were lowered while sodium and chlorine levels were elevated. These changes were accompanied by increased formation of ice crystal artifacts in myofibers. The findings suggest that trauma causes alterations in cytoplasmic macromolecules and in the state of water in the cells. The method of analysis provides a means for the further evaluation of antiinflammatory drugs.

Microprobe analysis of human masseter muscle biopsies.

Biopsies of the human masseter muscle were made using an intraoral approach. Sections were cut in a cryostat and freeze-dried. Using the myosin ATPase method at pH 9.3 the recognized heterogeneity of the muscle with respect to type I and type II fibers was confirmed. Sections were examined in an SEM by the method of energy dispersive X-ray analysis using a computer assisted semi-quantitative method (bulk analysis-thick sections) to determine the distribution of Na, P, S, Cl and K and their relative masses. Element concentrations in descending order were K, S, P, Na and Cl. Some element ratios were compared with results in the literature for other muscle (biochemical and microprobe analysis) and found to be generally in reasonable agreement. The results form a basis for the study of a prominent and accessible masticatory muscle in altered states.

Polymorphism of myosin light chains. An electrophoretic and immunological study of rabbit skeletal-muscle myosins.

Antibodies specific for rabbit fast-twitch-muscle myosin LCIF light chain were purified by affinity chromatography and characterized by both non-competitive and competitive enzyme-linked immunosorbent assay (ELISA) and a gel-electrophoresis-derived assay (GEDELISA). The antibodies did not cross-react with myosin heavy chains, and were weakly cross-reactive with the LC2F [5,5'-dithio-(2-nitrobenzoic acid)-dissociated] light chain and with all classes of dissociated light chains (LC1Sa, LC1Sb and LC2S), as well as with the whole myosin, from hind-limb slow-twitch muscle. The immunoreactivity of myosins with a truly mixed light-chain pattern (e.g. vastus lateralis and gastrocnemius) correlated with percentage content of fast-twitch-muscle-type light chains. A more extensive immunoreactivity was observed with diaphragm and masseter myosins, which were also characterized, respectively, by a relative or absolute deficiency of LC1Sa light chain. Furthermore, it was found that the LC1Sb light chain of masseter myosin is antigenically different from its slow-twitch-muscle myosin analogue, and is immunologically related to the LC1F light chain. Rabbit masseter muscle from its metabolic and physiological properties and the content, activity and immunological properties of sarcoplasmic-reticulum adenosine triphosphatase, is classified as a red, predominantly fast-twitch, muscle. Therefore our results suggest that the two antigenically different iso-forms of LC1Sb light chain are associated with the myosins of fast-twitch red and slow-twitch red fibres respectively.

Further studies on single fibres of bovine muscles.

Young & Davey (1981) (Biochem. J. 195, 317-327) identified numbers of polymorphs of myofibrillar proteins by sodium dodecyl sulphate/polyacrylamide gel electrophoresis of single muscle fibres isolated from three bovine muscles. Fibres were classed according to the distribution of polymorphs. The study has now been extended to eight diverse bovine muscles. The previous distinction made between fast and slow fibres is valid without exception in the extended study. Within these classes, variations in myofibrillar expression are examined within and between fibres, muscles and animals. Two slow muscles are contrasted; masseter is homogeneous in fibre type, whereas diaphragm is subtly heterogeneous, possibly arising from greater physiological demands. Of the myofibrillar polymorphs, attention is concentrated on two variants of fast-muscle myosin heavy chain. Both are present in all fast and mixed muscles examined, except sternomandibularis, and each is respectively associated with certain unidentified proteins. Within a muscle the fast-muscle myosin light-chain expression is the same irrespective of the heavy-chain variant. Histochemical techniques demonstrated that the variants are respectively associated with types IIA and IIB as defined by other investigators.