Mast cells and collagen fibrillogenesis.

This article presents 20 combinations of histochemical stainings for the determination of mast cell co-localization with the fibrous component of the connective tissue in the fibrillogenesis course. Best results were obtained using metachromatic detection of mast cells in combination with silver or picro-fuchsin impregnation, staining with brilliant green using van Gieson staining, and a combination of aniline blue staining with neutral red. Proposed variants of histochemical protocols open up new opportunities to analyze the participation of mast cells in extracellular matrix remodeling of the tissue microenvironment in the course of adaptive and pathological processes. Results obtained expand the current theoretical views of the process of fibrillogenesis in the extracellular matrix. They also shed new light on the participation of mast cell secretion components in the molecular mechanisms of fiber formation.

Tongue exercise and ageing effects on morphological and biochemical properties of the posterior digastric and temporalis muscles in a Fischer 344 Brown Norway rat model.

OBJECTIVE: This study sought to examin effects of age and tongue exercise on the posterior digastric (opener) and the temporalis (closer). We hypothesized 1) age would result in differing morphological (cross sectional area) and biochemical (myosin heavy chain isoform) components of these muscles; 2) tongue exercise would result in coactivation of these muscles inducing a decrease in age-related differences between age groups. DESIGN: Young adult (9 months) and old (32 months) Fischer 344 Brown Norway rats were randomized into a tongue exercise or control group. Post-training, posterior digastric and temporalis muscles were harvested and analyzed using: 1) Sodium Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis (SDS-PAGE) to assess percent myosin heavy chain (MyHC) content; 2) Immunohistochemical staining to determine cross sectional area (CSA). RESULTS: A larger proportion of slowly contracting MyHC isoforms in the posterior digastric and temporalis muscles were found in old. No significant main effects for age or exercise in fiber size were found in posterior digastric muscle. An interaction between age and exercise for temporalis cross sectional area indicated the old exercise group had smaller average cross sectional area than all other groups. CONCLUSIONS FINDINGS: suggest that: 1) Increasing age induces biochemical changes in muscles of the jaw, specifically showing an increase the proportion of slower contracting MyHC isoforms; 2) Increasing age and tongue exercise induce a reduction in muscle fiber cross sectional area in the temporalis muscle only. However, continued study of these cranial muscle systems is warranted to better understand these changes that occur with age and exercise.

A novel CIC-FOXO4 gene fusion in undifferentiated small round cell sarcoma: a genetically distinct variant of Ewing-like sarcoma.

Differential diagnosis of small round cell sarcomas (SRCSs) grouped under the Ewing sarcoma family of tumors (ESFT) can be a challenging situation for pathologists. Recent studies have revealed that some groups of Ewing-like sarcoma show typical ESFT morphology but lack any EWSR1-ETS gene fusions. Here we identified a novel gene fusion, CIC-FOXO4, in a case of Ewing-like sarcoma with a t(X;19)(q13;q13.3) translocation. The patient was a 63-year-old man who had an asymptomatic, 30-mm, well-demarcated, intramuscular mass in his right posterior neck, and imaging findings suggested a diagnosis of high-grade sarcoma. He was treated with complete resection and subsequent radiotherapy and chemotherapy. He was alive without local recurrence or distant metastasis 6 months after the operation. Histologic examination revealed SRCS with abundant desmoplastic fibrous stroma suggesting a desmoplastic small round cell tumor. Immunohistochemical analysis showed weak to moderate and partial staining for MIC2 (CD99) and WT1, respectively. High-throughput transcriptome sequencing revealed a gene fusion, and the genomic rearrangement between the CIC and FOXO4 genes was identified by fluorescence in situ hybridization. Aside from the desmoplastic stroma, the CIC-FOXO4 fusion sarcoma showed morphologic and immunohistochemical similarity to ESFT and Ewing-like sarcomas, including the recently described CIC-DUX4 fusion sarcoma. Although clinicopathologic analysis with additional cases is necessary, we conclude that CIC-FOXO4 fusion sarcoma is a new type of Ewing-like sarcoma that has a specific genetic signature. These findings have important implications for the differential diagnosis of SRCS.

Absence of developmental and unconventional myosin heavy chain in human suprahyoid muscles.

INTRODUCTION: Contradictory reports of the myosin heavy chain (MHC) composition of adult human suprahyoid muscles leave unresolved the extent to which these muscles express developmental and unconventional MHC. METHODS: By immunohistochemistry, separation sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE)-Coomassie, separation SDS-PAGE-Western blot, and mRNA PCR, we tested for conventional MHCI, MHCIIA, MHCIIX, developmental MHC embryonic and MHC neonatal, and unconventional MHC alpha-cardiac, MHC extraocular, and MHC slow tonic in adult human anterior digastric (AD), geniohyoid (GH), and mylohyoid (MH) muscles. RESULTS: By separation SDS-PAGE-Coomassie and Western blot, only conventional MHC are present. By immunohistochemistry all muscle fibers are positive for MHCI, MHCIIA, or MHCIIX, and fewer than 4 fibers/mm(2) are positive for developmental or unconventional MHC. By PCR, mRNA of MHCI and MHCIIA dominate, with sporadically detectable MHC alpha-cardiac and without detectable mRNA of other developmental and unconventional MHC. CONCLUSIONS: We conclude that human suprahyoid muscles AD, GH, and MH are composed almost exclusively of conventional MHC isoforms.

Age-related changes in rat genioglossus, geniohyoid and masseter muscles.

OBJECTIVES: The aim of this study was to elucidate age-related changes from adult to middle age in the contractile properties of the masseter, genioglossus and geniohyoid muscles of the rat. MATERIALS AND METHODS: We analysed the expressions of myosin heavy chain (MyHC) mRNAs and proteins as indicators of the contractile properties in these muscles obtained from rats at 6, 12, 18 and 24 months of age using real-time PCR and SDS-PAGE. RESULTS: We found no marked age-related changes in the expressions of MyHC mRNAs and proteins in rat masseter and geniohyoid muscles, suggesting that the biological ageing process does not affect contractile properties in these muscles. However, we found a decrease in the expression of MyHC IIb mRNA with ageing in the rat genioglossus muscle, suggesting that biological ageing process induces at least some fast-to-slow myofibre phenotype transition. CONCLUSION: The biological ageing process from adult to middle age appears to differentially affect different types of craniofacial muscles.

Protein differences between human trapezius and vastus lateralis muscles determined with a proteomic approach.

BACKGROUND: The trapezius muscle is a neck muscle that is susceptible to chronic pain conditions associated with repetitive tasks, commonly referred to as chronic work-related myalgia, hence making the trapezius a muscle of clinical interest. To provide a basis for further investigations of the proteomic traits of the trapezius muscle in disease, two-dimensional difference gel electrophoresis (2D-DIGE) was performed on the healthy trapezius using vastus lateralis as a reference. To obtain as much information as possible from the vast proteomic data set, both one-way ANOVA, with and without false discovery rate (FDR) correlation, and partial least square projection to latent structures with discriminant analysis (PLS-DA) were combined to compare the outcome of the analysis. RESULTS: The trapezius and vastus lateralis showed significant differences in metabolic, contractile and regulatory proteins, with different results depending on choice of statistical approach and pre-processing technique. Using the standard method, FDR correlated one-way ANOVA, 42 protein spots differed significantly in abundance between the two muscles. Complementary analysis using immunohistochemistry and western blot confirmed the results from the 2D-DIGE analysis. CONCLUSIONS: The proteomic approach used in the present study combining 2D-DIGE and multivariate modelling provided a more comprehensive comparison of the protein profiles of the human trapezius and vastus lateralis muscle, than previously possible to obtain with immunohistochemistry or SDS-PAGE alone. Although 2D-DIGE has inherent limitations it is particularly useful to comprehensively screen for important structural and metabolic proteins, and appears to be a promising tool for future studies of patients suffering from chronic work related myalgia or other muscle diseases.

Changes in the lingual muscles of obese rats induced by high-fat diet feeding.

OBJECTIVE: To elucidate the influences of obesity on the properties and volume of lingual (genioglossus and geniohyoid) muscles in obese rats. METHODS: We analysed the accumulation of triacylglycerol and the diameter of myofibres in the lingual muscles using histochemistry, and the MyHC composition using real-time PCR in rats fed a high-fat diet for 10 weeks. In the genioglossus and geniohyoid muscles, the percentage of oil droplet areas in the obesity group were 3.6 and 2.5 times greater than those in the control group, respectively (p<0.025). The diameters of the slow myofibres in the genioglossus and geniohyoid muscles were approximately 20% greater in the obesity group than in the control group (p<0.0001), while that of the fast myofibres in the geniohyoid muscle was approximately 10% greater in the obesity group than in the control group (p<0.0001). No significant difference in the expressions of any of the MyHC isoforms studied was found in any of the muscles studied between the obesity and control groups. CONCLUSION: High-fat diet feeding induced the fat deposition in the myofibre and influenced the structure of the lingual (genioglossus and geniohyoid) muscles.

Effects of increased occlusal vertical dimension on daily activity and myosin heavy chain composition in rat jaw muscle.

Mammalian skeletal muscles change their contractile-protein phenotype in response to mechanical loading and/or chronic electrical stimulation, implying that the phenotypic changes in masticatory muscles might result from new masticatory-loading conditions. To analyze the effects of increased occlusal vertical dimension (OVD) on daily activities and fibre-type compositions in jaw muscles, we measured the total duration of daily activity (duty time) and the myosin heavy chain (MyHC) compositions in the masseter and digastric muscles of freely moving control and bite-opened rats. In the control state, the duty time of the digastric muscle was higher than that of the masseter muscle at activity levels exceeding 5 and 20% of the day's peak activity. The opposite was true at activity levels exceeding 50 and 80% of the day's peak activity. The MyHCs consisted of a mixture of fast and slow types in the digastric muscle. The masseter consisted of mostly fast-type MyHC. The increment of OVD increased not only the duty time at activity levels exceeding 5, 20, 50 and 80% of the day' peak activity in both muscles but also the proportion of MyHC IIa in the masseter muscle and MyHC I in the digastric muscle at the expense of that of MyHC IIb. These results suggest that the increment of OVD changes masseter and digastric muscles towards slower phenotypes by an increase in their daily activities.

X-ray microanalysis of elements in the masticatory muscle after paresis of the right masseter.

Muscle activity and function appear to be related to ionic concentrations in the muscle. We investigated whether muscle paresis induced by injection of Botulinum toxin A (Botox) in 16-week-old pigs over a 56-day period is associated with ionic changes in the affected muscles. Tissue samples were taken from the masseter, temporalis, medial pterygoid, and geniohyoid muscles by a standardized method and used for energy-dispersive x-ray microanalysis in an environmental scanning electron microscope. The largest increase in Na(+) was measured in the right and left sides of the masseter muscle in treated animals. Additionally, a significant elevation of Na(+) was measured in the anterior part of the temporalis muscle and in the pterygoid muscle (P < 0.05). In temporalis and pterygoid muscles, an increase in sulfur in both sides of treated pigs' heads was observed. Botox((R)) has an indirect impact on ion concentrations, resulting in changes in muscle functional capacity and adaptive compensation of paretic muscle function by other muscles.

(90)Sr, (210)Po and (210)Pb in lichen and reindeer in Norway.

Concentrations of (90)Sr, (210)Po and (210)Pb in lichen and reindeer were studied in central (Østre Namdal) and southern Norway (Vågå) during 2000-2003. The study focussed on potential differences in concentrations of these nuclides in reindeer of different ages. Concentrations of (90)Sr in bones of approximately 10 year old adult females were about 40% higher than those in calves' antlers ((90)Sr concentrations in antlers and bones of calves are similar), while the available data from Vågå suggest that (90)Sr concentrations in reindeer calves decreased with an effective ecological half-time of 9.03+/-0.06 years during 1988-2002. Furthermore, (90)Sr concentrations were 50-80% higher in bone of reindeer of a similar age from Vågå compared to those from Østre Namdal. Concentrations of (210)Po and (210)Pb in muscle and liver tissues were comparable to those reported for reindeer in other Nordic areas, with no significant difference in (210)Po and (210)Pb concentrations between adults and calves or between reindeer from the two different study areas.

Defective neuromuscular synapses in mice lacking amyloid precursor protein (APP) and APP-Like protein 2.

Biochemical and genetic studies place the amyloid precursor protein (APP) at the center stage of Alzheimer's disease (AD) pathogenesis. Although mutations in the APP gene lead to dominant inheritance of familial AD, the normal function of APP remains elusive. Here, we report that the APP family of proteins plays an essential role in the development of neuromuscular synapses. Mice deficient in APP and its homolog APP-like protein 2 (APLP2) exhibit aberrant apposition of presynaptic marker proteins with postsynaptic acetylcholine receptors and excessive nerve terminal sprouting. The number of synaptic vesicles at presynaptic terminals is dramatically reduced. These structural abnormalities are accompanied by defective neurotransmitter release and a high incidence of synaptic failure. Our results identify APP/APLP2 as key regulators of structure and function of developing neuromuscular synapses.

Myosin heavy-chain isoform composition of human single jaw-muscle fibers.

Diversity in muscle contractile properties is based on the variability of contractile properties of single muscle fibers which in turn is related to the presence of different myosin heavy-chain (MyHC) isoforms. Human jaw muscles are featured by many hybrid fibers expressing more than one MyHC isoform. The purpose of this study was to determine the proportion of each isoform within these fibers for evaluation of the fiber's capacity of producing a large diversity in contractile properties. Electrophoretic separation of MyHC isoforms was performed on 218 single fibers of the temporalis and digastric muscles. Of these fibers, 100 were classified as hybrid fibers. Most hybrid fibers co-expressed MyHC-IIA and -IIX (n = 62); a smaller number co-expressed MyHC-I and -IIA (n = 14), MyHC-I and -IIX (n = 12), and MyHC-I, -IIA, and -IIX (n = 12). The proportions of the individual MyHC isoforms in the hybrid fibers varied highly, suggesting a large range of contractile properties among these fibers.

Intramuscular injection of antigens and adjuvant preferentially decreases 18:2n-6 and 18:3n-3 in pig neck muscle.

Linoleic (18:2n-6) and alpha-linolenic acids (18:3n-3) have many important physiological functions including immunomodulation. We tested how immunization influences the metabolism of 18:2n-6 and 18:3n-3 in the neck muscle of pigs. At 35 d old, pigs received either an intramuscular neck injection containing hen egg white lysozyme (HEWL), killed Mycobacterium tuberculosis, and Freund's complete adjuvant (immunized) or PBS (control). At 49 d old, immunized pigs received a booster injection of HEWL and Freund's incomplete adjuvant, and the control pigs received PBS into the neck. At 56 d old, all pigs received an intradermal injection of Mycobacterium bovis into the hind leg to induce a delayed-type hypersensitivity (DTH) reaction. At 57 d old, immunized pigs had a twofold increase in serum haptoglobin, a 10-fold increase in antibodies to HEWL, and the skinfold at the DTH reaction site was 10 times thicker than the controls. Both 18:2n-6 and 18:3n-3 (% composition) were approximately 25% lower in muscle TG, 40% lower in FFA, 50% lower in phospholipids, but not different in cholesteryl esters of the neck muscle of immunized pigs. The antigens in this model induce an increased response in the innate (haptoglobin), humoral (antibodies), and cellular (DTH) immune systems as well as a preferential decrease of 18:2n-6 and 18:3n-3 in the inflamed neck muscle. It appears that 18:2n-6 and 18:3n-3 are preferentially metabolized (possibly beta-oxidized) in response to antigens.

Differences in myosin heavy-chain composition between human jaw-closing muscles and supra- and infrahyoid muscles.

Jaw-closing muscles have architectural features suited to force production; supra- and infrahyoid muscles are better adapted to produce velocity and displacement. It was hypothesized that this difference in function would be reflected in myosin heavy-chain (MyHC) composition (equivalent to contraction velocity) and fibre-type cross-sectional area (equivalent to force). MyHC composition was determined in muscles obtained from eight human cadavers, using monoclonal antibodies against MyHC isoforms. Jaw closers contained 4.2 times fewer type IIA fibres and 5.2 times more hybrid fibres than suprahyoid muscles, and 3.9 times fewer type IIA fibres and 3.2 times more hybrid fibres than the infrahyoid muscles. In the jaw closers, MyHC-I was expressed in approx. 70% of all fibres (pure+hybrid), in the suprahyoid muscles in approx. 40%, and in the infrahyoid muscles in approx. 46%. In the jaw closers, type I fibres were 40% larger in diameter than in the supra- and infrahyoid muscles. It can be concluded that the jaw closers have characteristics of slow muscles, and that the supra-/infrahyoid muscles have characteristics of fast muscles.

Histological characteristics of sternoclavicular beta 2-microglobulin amyloidosis and clues for its histogenesis.

BACKGROUND: The pathogenesis of beta 2-microglobulin amyloidosis (A beta 2m) has yet to be fully elucidated. METHODS: We describe the distribution and extent of A beta 2m deposition and macrophagic infiltration in cartilage, capsule, and synovium of sternoclavicular joints obtained postmortem from 54 patients after 3 to 244 (median 46) months of dialysis. Twenty-four nonuremic patients served as a control group. The diagnosis of amyloidosis (A) rested on a positive Congo Red staining (typical birefringence) and that of A beta 2m on positive immunostaining of the A deposits with a monoclonal anti-beta 2m antibody. The size of A deposits was measured. RESULTS: A beta 2m was detected in 32 (59%), and non-beta 2m amyloid (Anon beta 2m) was detected in an additional 8 (15%) of the 54 dialyzed patients. A beta 2m deposits were present in the cartilage of all A beta 2m (+) patients (100%). They were localized solely in the cartilage in 27% of the cases, either as a thin patchy layer or as a continuous thicker layer (identified as stage I). A beta 2m was additionally present in the capsule and/or synovium without macrophages in 27% of the cases (identified as stage II). The correlation between the size of cartilaginous deposits and dialysis duration (P = 0.02) as well as with the prevalence (P = 0.03) and size of capsular deposits (P = 0.02) suggests that stage II is a later stage of A deposition. Clusters of macrophages were detected around capsular and synovial amyloid deposits in 46% of the cases (identified as stage III). The longer duration of dialysis in those with stage III as well as the relationship between the size of the A beta 2m deposits and the prevalence of macrophagic infiltration suggests that stage III is the last stage of A beta 2m deposition. Marginal bone erosions were observed in 9 out of 12 patients with stage III deposits. Their size was correlated with that of cartilaginous deposits (P = 0.01). Among the 24 control patients, Anon beta 2m was detected in 12 patients (cartilage 100%, capsule 8%, synovium 30%). CONCLUSIONS: The earliest stage of A beta 2m deposition occurs in the cartilage. A beta 2m subsequently extends to capsule and synovium. These two first stages do not require macrophage infiltration. Macrophages are eventually recruited around larger synovial or capsular deposits in the final stage. Marginal bone erosions develop in this late stage.

The NADPH-diaphorase staining reaction in relation to the aminergic and peptidergic nervous system and the musculature of adult Diphyllobothrium dentriticum.

A new category of signalling molecules--transmitter gases--has appeared. Nitric oxide (NO) is generated by nitric oxide synthase (NOS) and diffuses as a short-lived transcellular messenger through the plasma membrane. NADPH-diaphorase (NADPH-d) is a marker enzyme for NO-producing neurons. In this study the pattern of NADPH-d stained neurons in Diphyllobothrium dendriticum is described and compared to the pattern of aminergic and peptidergic neuronal elements and to that of the musculature stained with TRITC-labelled phalloidin. NADPH-d staining was observed in neurons in the bilobed brain and along the 2 main nerve cords and in nerve fibres close to the body musculature and the musculature of the reproductive ducts, the walls of the testicular follicles and in sensory endings in the tegument. The NADPH-d staining reaction and the 5-HT or FMRFamide immunoreactivities occur in separate sets of neurons.

Doxorubicin chemomyectomy as a treatment for cervical dystonia: histological assessment after direct injection into the sternocleidomastoid muscle.

The sternocleidomastoid muscle (SCM) is one of the major muscles involved in producing abnormal head position in cervical dystonia patients. This study tested whether doxorubicin chemomyectomy, direct injection of doxorubicin into the SCM to permanently remove muscle fibers, has the potential to be a nonsurgical, permanent treatment for cervical dystonia. The right SCM of rabbits was injected with either 1 or 2 mg doxorubicin. Animals were sacrificed 1-2 months postinjection. The SCM was prepared for histological examination of muscle fiber loss and fiber type composition. In all cases, direct injection of doxorubicin resulted in significant decreases in total muscle cross-sectional areas ranging from 75% up to 98%. Individual myofiber cross-sectional areas were smaller than normal after 2 mg doxorubicin treatment, but similar to normal fiber size after 1 mg doxorubicin. There were increased numbers of myofibers that expressed slow and neonatal myosin heavy chain isoforms in these remaining muscle fibers compared to the untreated SCM on the contralateral side. Developmental myosin heavy chain (MHC) was also present in 53% of the remaining myofibers of the treated muscles. The fiber type composition of muscles contralateral to the doxorubicin injections was compared to the fiber type composition of SCM from normal, untreated controls; no difference was seen in the proportions of fast, slow, and neonatal MHC fiber types in these SCM muscles. In summary, the direct injection of doxorubicin into the SCM resulted in significant muscle loss. This supports the use of doxorubicin chemomyectomy as a potential permanent, nonsurgical treatment for cervical dystonia.

In vivo lactate editing with simultaneous detection of choline, creatine, NAA, and lipid singlets at 1.5 T using PRESS excitation with applications to the study of brain and head and neck tumors.

Two T2-independent J-difference lactate editing schemes for the PRESS magnetic resonance spectroscopy localization sequence are introduced. The techniques, which allow for simultaneous acquisition of the lactate doublet (1.3 ppm) and edited singlets upfield of and including choline (3.2 ppm), exploit the dependence of the in-phase intensity of the methyl doublet upon the time interval separating two inversion (BASING) pulses applied to its coupling partner after initial excitation. Editing method 1, which allows for echo times TE = n/J (n = 1, 2, 3, . . . . ), alters the BASING carrier frequency for each of two cycles so that, for one cycle, the quartet is inverted, whereas, for the other cycle, the quartet is unaffected. Method 2, which also provides water suppression, allows for editing for TE > 1/J by alternating, between cycles, the time interval separating the inversion pulses. Experimental results were obtained at 1.5 T using a Shinnar Le-Roux-designed maximum phase inversion pulse with a filter transition bandwidth of 55 Hz. Spectra were acquired from phantoms and in vivo from the human brain and neck. In a neck muscle study, the lipid suppression factor, achieved partly through the use of a novel phase regularization algorithm, was measured to be over 10(3). Spectra acquired from a primary brain and a metastatic neck tumor demonstrated the presence of lactate and choline signals consistent with abnormal spectral patterns. The advantages and limitations of the methods are analyzed theoretically and experimentally, and significance of the results is discussed.

Adaptations of masticatory muscles to a hyperpropulsive appliance in the rat.

The plasticity of masticatory muscles was studied by comparing rats that were wearing a protrusive appliance and were kept on a liquid diet with two control groups: (1) pair-fed rats and (2) rats that had free access to ordinary pelleted food. The animals were 45 days old at the beginning of the experiment and were studied for a period of 20 days. Three jaw muscles with different functions were examined: masseter, temporalis, and digastric. Muscle fiber composition was determined (1) by fiber counting after staining with four monoclonal antibodies, which were able to recognize the four major myosin heavy chain (MHC) isoforms and therefore four fiber types (I, IIA, IIX, IIB) and (2) by electrophoresis on 6% polyacrylamide gels. The comparison between free-diet rats and pair-fed rats showed that the change from a hard pelleted diet to a liquid diet caused a shift in fiber type and MHC distribution, characterized by an increase of IIB MHC in temporalis and digastric muscles but not in the masseter muscle. The comparison between pair-fed rats and rats wearing appliances showed on the contrary a decrease in IIB MHC and an increase in IIA and IIX MHC. The results support the conclusions that (1) rat jaw muscles can quickly adapt to functional demand changing their fiber type composition, (2) the changes appear restricted inside the fast fiber population, and (3) fiber-type changes caused by dietary variation are not less than those caused by orthodontic intervention and must be taken into account to assess the effect of the appliance correctly.

Myosin expression in the jaw-closing muscles of the domestic cat and American opossum.

Sodium dodecylsulphate-polyacrylamide gel electrophoresis (SDS-PAGE), glycerol SDS-PAGE, two-dimensional electrophoresis, and protein immunoblotting techniques were used to identify myosin heavy chain (MHC) and light chain (MLC) isoforms in limb and masticatory muscles of the cat and American opossum. The fibre types in which these isoforms are expressed were identified by histochemistry and immunohistochemistry. Antibodies specific for the type IIM MHC isoform characteristic of cat jaw-closing muscles and the type I MHC isoform were produced and characterized. The IIM antibody stained the majority of fibres found in the jaw-closing muscles of both species. These IIM-containing fibres characteristically had a histochemical ATPase that remained active after both acid and alkali pre-incubations. A minority of type I fibres was also present in cat jaw-closing muscles, and these reacted positively with antibody specific for type I MHC. It was confirmed that the vast majority of fibres in the cat jaw-closing muscles contained only the characteristic masticatory MHC (IIM) and masticatory MLCs (LC1m and LC2m). These muscles did not contain either the type II fibre isoforms of limb muscles or the atrial cardiac (alpha-cardiac) MHC. The type IIM MHC could also be identified in jaw-closing muscles of the opossum. Two-dimensional gel electrophoresis was used to identify the MLC composition of single, histochemically defined, type I fibres in the cat soleus and deep masseter. The type I fibres of limb muscle contained the usual slow MLCs, but type I fibres from the jaw-closing muscles contained only the masticatory light chains.