The effects of increasing occlusal vertical dimension on the deep masseter of rat at different ages.

OBJECTIVE: To investigate the influence of increasing the occlusal vertical dimension (iOVD) on the fibre-type distribution and ultrastructure of deep masseter of rat at different ages. DESIGN: A total of forty-eight male Wistar rats were divided into two groups according to age: 'teenage' group (n=24, 1.5 months) and 'young adult' group (n=24, 8 months). Both the teenage and the young adult rats were then randomly divided into the control group (n=12) and the experimental group (n=12). The occlusal vertical dimensions of the rats in the experimental groups were increased by placing composite resin on all maxillary molars. The fibre-type distribution and ultrastructure of the deep masseter were subsequently observed on day 7 and day 14 after iOVD. RESULTS: In the teenage experimental group, the proportion of type IIa fibres increased, while the proportion of type IIb and type IIx fibres decreased by day 7 after iOVD (P<0.05). However, no significant fibre phenotype transformation was observed in the young adult experimental group until day 14 after iOVD. In addition, the proportion of type IIa in the teenage experimental group was higher than that of the young adult experimental group on day 7 and 14 (P<0.05). Under the transmission electron microscope, muscle fibre reconstruction and the compensatory increase in the number and volume of mitochondria appeared earlier in the teenage experimental group. The cellular traumatic reaction was less than that in the young adult experimental group. CONCLUSION: The teenage rat alters masseter muscle structure to a slower phenotype earlier and to a greater degree than that of the young adult rat when increasing the occlusal vertical dimension.

Metabolic Changes in Masseter Muscle of Rats Submitted to Acute Stress Associated with Exodontia.

Clinical evidence has shown that stress may be associated with alterations in masticatory muscle functions. Morphological changes in masticatory muscles induced by occlusal alterations and associated with emotional stress are still lacking in the literature. The objective of this study was to evaluate the influence of acute stress on metabolic activity and oxidative stress of masseter muscles of rats subjected to occlusal modification through morphological and histochemical analyses. In this study, adult Wistar rats were divided into 4 groups: a group with extraction and acute stress (E+A); group with extraction and without stress (E+C); group without extraction and with acute stress (NO+A); and control group without both extraction and stress (NO+C). Masseter muscles were analyzed by Succinate Dehydrogenase (SDH), Nicotinamide Adenine Dinucleotide Diaphorase (NADH) and Reactive Oxygen Species (ROS) techniques. Statistical analyses and two-way ANOVA were applied, followed by Tukey-Kramer tests. In the SDH test, the E+C, E+A and NO+A groups showed a decrease in high desidrogenase activities fibers (P < 0.05), compared to the NO+C group. In the NADH test, there was no difference among the different groups. In the ROS test, in contrast, E+A, E+C and NO+A groups showed a decrease in ROS expression, compared to NO+C groups (P < 0.05). Modified dental occlusion and acute stress--which are important and prevalent problems that affect the general population--are important etiologic factors in metabolic plasticity and ROS levels of masseter muscles.

Expression of sarcoplasmic-endoplasmic reticulum Ca-ATPase isoforms in masticatory muscles.

The aim of this study was to characterize the sarcoplasmic-endoplasmic reticulum Ca-ATPase (SERCA) isoforms in rabbit masticatory muscles compared with those in fast-twitch muscle. It was hypothesized that combined expression of the SERCA isoforms in fast- and slow-twitch muscles accounts for lower Ca-ATPase activity. SERCA was isolated by differential centrifugation, the isoforms were determined by ELISA, and the activity of each isoform was measured using a colorimetric method. Activity was tested for significance by anova, and the distribution of isoforms was assessed using the chi-square test (P < 0.05) and correlated to SERCA activity using Spearman's rank correlation. SERCA1 was predominant (90.5%) in fast-twitch muscle, whereas a mixture of SERCA isoforms was found in masticatory muscles: 62-78% was SERCA2, 20-37% was SERCA1, and the SERCA3 content was negligible. Depressor muscles showed a significantly higher content (77.8%) of SERCA2, and elevator muscles showed a higher content (35.4%) of SERCA1. Elevator muscles showed higher expression of SERCA2a (58%), and depressor muscles showed higher expression of SERCA2b (20%). The SERCA1 content was mainly SERCA1a and significantly higher for elevator muscles (33%), whereas depressor muscles showed a higher content of SERCA1b (4%). The SERCA1 content of fast-twitch muscle was mainly SERCA1a (88.5%). It is concluded that the mixture of different SERCA isoforms, along with a substantial content of SERCA2b, in masticatory muscles would support lower Ca-ATPase activity and calcium transport.

Effects of low-level laser therapy on the oxidative metabolism and matrix proteins in the rat masseter muscle.

OBJECTIVE: This study aims to analyze the effects of low-level laser therapy (LLLT) on the oxidative activity and the expression/activity of metalloproteinases of the masseter muscle. BACKGROUND DATA: Currently in dentistry LLLT has been used on patients with muscular disorders, such as the temporomandibular disorders (TMDs) but its effect at the cellular level has not been fully elucidated. METHODS: Thirty male Wistar rats divided into 6 groups (n=5) received 10 laser irradiations (780 nm, 5 mmW, CW laser, illuminated area 0.04 cm(2), power density 125 mW/cm(2)), with different energy densities (group I-0; group II-0.5; group III-1.0; group IV-2.5; group V-5.0; and group VI-20 J/cm(2)). Muscles were processed for nicotinamide adenine dinucleotide diaphorase (NADH) and sucinate dehydrogenase (SDH) activities and zymography. The photomicrographs were evaluated by the point counting method using a test system and ImageJ software; and by the ANOVA statistical test. The proteinases' secretion/activity was qualitatively analyzed by zymography. RESULTS: LLLT significantly increased (p<0.05) masseter muscle oxidative metabolism shown by the increased area of intermediary fibers in the NADH (groups IV, V, and VI) and SDH (group V) reactions. The same metabolic pattern was observed among the groups in both reactions. Sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) zymography detected only the MMP-2 expression/activity for the untreated-control group (group I). The exposure to LLLT increased the activity of MPP-2 in group VI and the activity of MMP-9 in all groups exposed to different energy densities of laser irradiation (groups II, III, IV, V, and VI). CONCLUSIONS: Thus, LLLT stimulated the oxidative metabolism and the expression of matrix metalloproteinase (MMPs) of the masseter muscle, which may indicate a matrix remodeling process. However, group VI did not show the best results for oxidative metabolism, probably indicating that the dosage they were given was high for this protocol.

[Effect of radiation on the activity and expression of Ca(2+)-Mg(2+)-ATPase in rat masseter muscle].

OBJECTIVE: To investigate the activity and expression of Ca(2+)-Mg(2+)-ATPase in irradiated rat masseter muscle. METHODS: The rats were irradiated locally with a single dose of 20 Gy X-ray. The activities of Ca(2+)-Mg(2+)-ATPase were measured with colorimetric method. The protein expression of Ca(2+)-Mg(2+)-ATPase was determined by Western blotting and immunohistochemistry. RESULTS: The activities of Ca(2+)-Mg(2+)-ATPase in masseter muscle decreased by approximately 20% and 40% in irradiated rats on days 3 and 30 postirradiation. There was significant difference in the expression of Ca(2+)-Mg(2+)-ATPase protein between irradiated and nonirradiated rats on day 30 postirradiation. Ca(2+)-Mg(2+)-ATPase protein was found in the cytoplasm of masseter muscle. CONCLUSIONS: The decrease of ATPase activity played an important role in the cause of radiation-induced skeletal muscle injury, while there was no significant reduction in the expression of Ca(2+)-Mg(2+)-ATPase protein in irradiated rat masseter muscle.

Indices of extracellular matrix turnover in human masseter muscles as markers of craniofacial form--a preliminary study.

Environmental remodelling of the craniofacial musculature is obligatory for successful outcomes following interventions such as functional appliance therapy or orthognathic surgery. Genetically driven remodelling of the craniofacial musculature is also seen in individuals with altered facial form. The processes that are involved in the remodelling of intramuscular connective tissue need to be activated in such situations. Such processes require activity of matrix metalloproteinases (MMPs) and the tissue inhibitors of matrix metalloproteinases (TIMPs), which are responsible for extracellular matrix (ECM) turnover. The aim of this study was, therefore, to establish the expression of MMP-2 and MMP-9 and their inhibitors, TIMP-1 and TIMP-2, in the masseter muscle of humans with both normal and increased vertical facial form and to assess whether this expression had any value as a predictor of facial form. Biopsies were taken from 20 subjects (10 with vertical facial deformity and 10 with normal vertical facial form to act as a control group). The sample group consisted of 15 females and 5 males and the average age of the donors +/- standard deviation (SD) was 26.04 +/- 6.16 years (range: 17.67-31.25 years). Biopsy samples were then subjected to zymography and reverse zymography to assess MMP and TIMP expression, respectively. Lateral skull cephalograms were analysed for each subject using Spearman's rho correlation coefficients and Mann-Whitney U-tests. TIMP-1 activity was consistently expressed in human masseter muscle. MMP-2, MMP-9, and TIMP-2 activity, when detected, was at a low level. These data indicate that in most individuals, an excess of TIMP-1, compared with MMP-2 and MMP-9, limits ECM turnover in human masseter muscle. There was a demonstrable variation in proteinase expression between different individuals. These preliminary findings, however, do not confirm that indices of ECM turnover are a reflection of an individual's vertical facial form.

Activation of caspase 3, 9, 12, and Bax in masseter muscle of mdx mice during necrosis.

The mdx mouse, a model of muscular dystrophy, lacks dystrophin, a cell membrane protein. It is known that the lack of dystrophin causes muscle fiber necrosis from 2 weeks after birth, and the majority of necrotic muscle fibers are replaced by regenerated muscle fibers by 4 weeks after birth. A recent study indicated the possibility that mitochondria-mediated intracellular stress, a phenomenon similar to apoptosis, may be produced during muscle fiber necrosis, but did not analyze endoplasmic reticulum-mediated intracellular stress. Therefore, we examined the expression of the caspase-12 gene involved in the endoplasmic reticulum stress pathway and the Bax, caspase-9, and caspase-3 genes involved in the mitochondrial stress pathway in the mdx masseter muscle. We found over-expression of caspase-12 in cells at 2-3 weeks after birth when muscle fiber necrosis was not prominent. This suggests that stress occurs in the endoplasmic reticulum to maintain cell morphology in the absence of dystrophin. In addition, Bax was abundantly expressed in the mdx masseter muscle at 3 weeks after birth, and the expression of caspase-9 and -3 was prominent at 3-4 weeks after birth when necrosis and regeneration were marked. These results indicate that endoplasmic reticulum and mitochondrial stresses are produced during necrosis of the mdx masseter muscle, and suggest that these events are a phenomenon similar to apoptosis.

Effect of changes in food consistency on NADH-ubiquinone oxidoreductase activity and levels of mRNA for ND1, 51kDa, 75kDa and myosin heavy chain isoforms in two different portions of rat masseter muscle.

We investigated the effect of a change in food consistency on properties of the masseter muscle in 3-week-old rats fed a soft diet for 9 weeks (Group S) and fed a soft diet for 5 weeks followed by a hard diet for 4 weeks (Group S-H). The NADH-O2 oxidoreductase activity, levels of mRNAs transcribed from genes encoding NADH-ubiquinone oxidoreductase (Complex I: ND1, 51kDa, and 75kDa) and myosin heavy chain (MyHC) isoforms and the phenotype of the muscle fibers were measured in the superficial and deep portions of the muscle. In the period from 8 weeks to 12 weeks of age, NADH-O2 oxidoreductase enzyme activity in both the superficial and deep portions of the muscle showed similar patterns in Group S and Group S-H. In contrast, the ND1, 51kDa and 75kDa mRNA levels in the superficial and deep portions of the masseter muscle in the Group S-H were higher than those of Group S in the 12-week-old rats, except for the 51kDa mRNA in the superficial portion of the masseter muscle. MyHC-IIa and MyHC-IId/x mRNA levels in the superficial portion of the masseter muscle were higher in the Group S-H than in the Group S. These observations suggest that short-term feeding stress such as the transition from a soft diet to a hard diet causes changes in oxidative metabolism, in mRNA levels for the Complex I components ND1 and 75kDa, and the mRNA levels for the MyHC isoforms IIa and IId/x in the superficial portion of rat masseter muscle, but no changes in the composition of muscle fiber types.

Alterations in enzyme histochemical characteristics of the masseter muscle caused by long-term soft diet in growing rabbits.

OBJECTIVE: Recently young people have an increasing tendency to intake an easily chewable diet and spend less time on mastication. The aim of the present study was to investigate the histochemical effects of long-term soft diet on the masseter muscle in growing rabbits. MATERIALS AND METHODS: Twelve young male Japanese white rabbits were divided into two groups (n = 6 each) at weaning (1 month after birth) and fed a solid diet (control group) or a powder diet (soft-diet group). The duration of the experimental period was 6 months. Masseter fibers from the superficial and the deep portions were histochemically defined as type 1, 2A, 2B, or 2C fibers. RESULTS: As compared with that of the control, the deep masseter of the soft-diet group showed a significantly lower ratio of type 1 fiber cross-sectional area to total area (6.3 and 10.1% for the soft-diet and control group, respectively), significantly more type 2A fibers (74.0%vs 50.3%) and significantly fewer type 2B fibers (4.3%vs 12.5%). However, fiber size did not differ between the two groups. NADH-tetrazolium-reductase (NADH-TR) of the masseter was less reactive in the soft-diet group, reflecting a lower oxidative capacity. CONCLUSIONS: These findings indicate that the alteration of the functional activities contributed to selective disuse influences on the type 1 and type 2B fibers, and a resultant increase in type 2A fibers. This study suggests that long-term alteration of jaw function induced by a soft diet can lead to adaptations of the masseter muscle.

Weaning affects lipoprotein lipase activity and gene expression in adipose tissues and in masseter but not in other muscles of the calf.

The nutritional and physiological modifications that occur during the weaning period induce adaptations of tissue metabolism in all mammal species. Among the adaptations due to weaning in ruminants, the regulation of lipoprotein lipase (LPL) activity, one of the rate-limiting steps of fatty acid utilization by tissues, was still unknown. The present study aimed at comparing LPL activity and gene expression in the heart, seven skeletal muscles and three adipose tissue sites between two groups of seven preruminant (PR) or ruminant (R) calves having a similar age (170 d), similar empty body weight (194 kg) at slaughter, and similar net energy intake from birth onwards. Triacylglycerol content of adipose tissues was 16 % lower in R than in PR calves, This could be partly the result from a lower LPL activity (-57 %, ). LPL mRNA levels were also lower in R calves (-48 % to -68 %, ) suggesting a pretranslational regulation of LPL activity. Activity and mRNA levels of LPL did not differ significantly in the heart and skeletal muscles except in the masseter in which LPL activity and mRNA levels were higher (+50 % and +120 % respectively, ) in the R calves. Regulation of LPL in masseter could be explained by the high contractile activity of this muscle after weaning due to solid food chewing. In conclusion, weaning in the calf affects LPL activity and expression in adipose tissues, but not in skeletal muscles except the masseter.

Histopathologic features of masseter muscle in the distrophic hamster (UM-X7.1 Syrian hamster).

Dystrophic hamster has been regarded as the useful model animal for Severe childhood autosomal recessive muscular dystrophy (SCARMD). Although, many studies on Dystrophic hamster have utilized the muscular tissue of the trunk, however no study have been analyzed for the masticatory muscle. For this study, we used a Dystrophic hamster (UM-X7.1 Syrian hamster) to histochemically investigate the effect of muscular dystrophy on the masseter muscle. Large and small regenerated muscle fibers, and necrotic fibers were detected almost in all areas. Opaque fiber, hypertrophic fiber with fiber splitting structure and necrotic fiber filled up by mononuclear phagocytes were recognized. The region, in which the mononuclear phagocytic cells infiltrated, showed strong positivity to acid phosphatase, and lysosome enzyme. There were many muscle fibers with reduced levels of succinate dehydrogenase (SDH) activities in the muscle fiber. Some TUNEL-positive cells were confirmed in both necrotic and non-necrotic areas. It was suggested that a part of TUNEL-positive cells are the cells originated from the connective tissue or immunocytes. In this result, histopathologic changes of the masseter muscle of the UM-X7.1 Syrian hamster was similar to muscle of the body trunk in the past reports. As the result, it was suggested that jaw closing movements may be negatively affected caused by the decline of the masseter muscle twitch. And, the point of view by which apoptosis is the trigger for the muscle fiber collapse were not seen in the Dystrophic hamster masseter muscle. We suggest that apoptosis is a one step in the process of regeneration of muscle fibers.

Elevation of histidine decarboxylase activity in skeletal muscles and stomach in mice by stress and exercise.

The effects of different types of stress (water bathing, cold, restraint, and prolonged walking) on histidine decarboxylase (HDC) activity in masseter, quadriceps femoris, and pectoralis superficial muscles, and in the stomach were examined in mice. All of these stresses elevated gastric HDC activity. Although water bathing, in which muscle activity was slight, was sufficiently stressful to produce gastric hemorrhage and to increase gastric HDC activity, it produced no detectable elevation of HDC activity in any of the muscles examined. The other stresses all elevated HDC activity in all three muscles. We devised two methods of restraint, one accompanied by mastication and the other not. The former elevated HDC activity in the masseter muscle, but the latter did not. These results suggest that 1) HDC activity in the stomach is an index of responses to stress, 2) the elevation of HDC activity in skeletal muscles during stress is induced partly or wholly by muscle activity and/or muscle tension, and 3) stress itself does not always induce an elevation of HDC activity in skeletal muscles.

Coordinate expression of Ca2+-ATPase slow-twitch isoform and of beta calmodulin-dependent protein kinase in phospholamban-deficient sarcoplasmic reticulum of rabbit masseter muscle.

Modulation of sarcoplasmic reticulum (SR) Ca(2+) transport by endogenous calmodulin-dependent protein kinase II (CaM K II) involves covalent changes of regulatory protein phospholamban (PLB), as a common, but not the only mechanism, in limb slow-twitch muscles of certain mammalian species, such as the rabbit. Here, using immunofluorescent techniques in situ, and biochemical and immunological methods on the isolated SR, we have demonstrated that rabbit masseter, a muscle with a distinct embryological origin, lacks PLB. Accommodating embryological heterogeneity in the paradigm of neural-dependent expression of specific isogenes in skeletal muscle fibers, our results provide novel evidence for the differential expression in the SR of 72 kDa beta components of CaM K II, together with the expression of a slow-twitch sarcoendoplasmic reticulum Ca(2+)-ATPase isoform, both in limb muscle and in the masseter.

Identification of matrix metalloproteinases and their tissue inhibitors type 1 and 2 in human masseter muscle.

Changes in masticatory muscle structure and function are either developmental, as seen in anomalies of facial form, or adaptive, as seen during procedures such as orthognathic surgery and functional-appliance orthodontic therapy. Remodelling of muscle extracellular matrix is pivotal in these processes. This turnover is mediated via members of the family of enzymes known as matrix metalloproteinases (MMP) and inhibited by the tissue inhibitors of metalloproteinases (TIMP). The aim here was to investigate the in vivo pattern of expression and distribution of MMPs and TIMPs in masseter muscle of humans with both normal and abnormal facial forms. Masseter muscle biopsies were taken from 10 patients, four with long-face syndrome and six normal controls as confirmed by cephalometry. Immunohistochemical techniques were used to show the pattern and distribution of MMPs and TIMP proteins in the muscle. Zymography of tissue extracts was used to determine the presence of MMP activity. Reverse transcriptase-polymerase chain reaction (RT-PCR) was used to detect the presence of MMP and TIMP-2 mRNA. MMP-1 was expressed around the individual muscle fibres, especially in those fibre surfaces in contact with the interstices of the connective tissue and around blood vessels. MMP-9 staining was less intense and was expressed in the interstices of the connective tissue and around blood vessels. Zymography of protein extracts confirmed that MMP-9 activity was present. MMP-2 and MMP-3 were not expressed in the samples, although MMP-2 mRNA could be detected by RT-PCR and its activity could be detected by zymography. Intense TIMP-1 staining was present around each muscle fibre, in the interstices of the connective tissue and surrounding blood vessels; TIMP-2 mRNA could be detected in all samples. These staining patterns were seen in all biopsies examined and were irrespective of the facial form of the donor. These findings provide evidence that the mechanisms required for matrix remodelling are present in the human masseter muscle.

Possible involvement of histamine in muscular fatigue in temporomandibular disorders: animal and human studies.

As an approach to clarifying the molecular basis of pain and fatigue in muscles involved in temporomandibular disorders, we examined the activity of histidine decarboxylase (HDC), the enzyme which forms histamine, in the masseter muscles of mice. In the resting muscle, HDC activity was very low. Direct electrical stimulation of the muscle markedly elevated HDC activity. HDC activity rose within 3 hrs of the electrical stimulation, peaked at 6 to 8 hrs, and then gradually declined. Intraperitoneal injection of a small amount of interleukin-1 (IL-1) (from 1 to 10 microg/kg) produced a similar elevation of HDC activity in the masseter muscle. We also examined the effect of an antihistamine, chlorphenylamine (CP), on temporomandibular disorders in humans and compared it with that of an anti-inflammatory analgesic, flurbiprofen (FB). Two groups received one or the other of the drugs daily for 7 days, and they were asked about their signs and symptoms before and after the treatment. A positive evaluation of their treatment was made by 74% of the CP group, but by only 48% of the FB group. Although the effects of CP on the limitation of mouth-opening and on joint noise were negligible, about 50% of the CP group answered positively concerning the drug's effect on spontaneous pain or pain induced by chewing or mouth-opening. The positive evaluation for CP (50%) in relieving associated symptoms (headache or shoulder stiffness) was significantly greater than for FB (13%). FB showed effectiveness similar to but sometimes weaker than that of CP on several symptoms. On the basis of these and previous results and the known actions of histamine, we propose that the histamine newly formed following the induction of HDC activity, which is itself mediated by IL-1, may be involved in inducing pain and, possibly, stiffness in muscles in temporomandibular disorders.

Inhibition of skeletal sarcoplasmic reticulum Ca2+-ATPase activity by deferoxamine nitroxide free radical.

Deferoxamine is an inhibitor of iron-dependent free radical reactions. Despite the antioxidant roles, prolonged clinical use of the chelator is far from benign, and paradoxically, deferoxamine has been shown to promote lipid peroxidation. The possible toxicity of the drug's metabolites, such as deferoxamine nitroxide free radical, deserves attention. We, therefore, tested the hypothesis that deferoxamine nitroxide radicals produced as a result of enzymatic one-electron oxidation of deferoxamine by horseradish peroxidase in the presence of H2O2 are capable of inactivating Ca2+-ATPase of skeletal sarcoplasmic reticulum microsomes as a model system with which to explore the effect of the radical on a biological membrane. Ca2+-ATPase activity of sarcoplasmic reticulum was depressed by exposure to Fenton's reagent (H2O2/FeSO4); the observed effect was significantly enhanced by deferoxamine. We found that the Fenton reaction produced hydroxyl radical, as determined by electron spin resonance spectroscopy. The formation of hydroxyl radical was completely inhibited by deferoxamine; instead, under the same experimental conditions (in the presence of sarcoplasmic reticulum vesicles with or without FeSO4 but without spin trap 5, 5-dimethyl-1-pyrroline N-oxide), the spectral shape and hyperfine coupling constants of electron spin resonance signals confirmed to be long-lived deferoxamine radical were obtained. Furthermore, exposure of sarcoplasmic reticulum vesicles to deferoxamine radical formed by horseradish peroxidase via reaction with H2O2 caused an inhibition of the Ca2+-ATPase activity. The findings show that the sarcoplasmic reticulum vesicles can act as peroxidases and suggest that deferoxamine enhances the decreased Ca2+-ATPase activity afforded by H2O2/FeSO4 due to formation of its metabolites, possibly deferoxamine nitroxide free radical.

Oxidative capacity of rat masseter muscle after implantation of thyrotropin-releasing hormone microspheres in proximity to trigeminal motoneurones.

Earlier work has shown that two important consequences of implanting thyrotropin-releasing hormone (TRH) microspheres near motoneurones within the trigeminal motor nucleus of actively growing rats are increased muscle mass and a darkening of the implant-side masticatory muscles. These phenomena have been associated with altered neuromuscular activity patterns and biomechanical forces that directly influence craniofacial growth and development. Now, whether the implantation of TRH microspheres in proximity to trigeminal motoneurones would affect the oxidative capacity of the implant-side masseter muscles was investigated. Cytochrome C oxidase (COX) assays were carried out for both implant- and non-implant-side masseters of TRH (n = 5) and blank microsphere (n = 6) Sprague-Dawley rats after stereotactic surgery at 35 days of age. Analyses of both groups at 14 days post-implantation revealed that the COX activity levels of implant-side masseters in TRH-implanted rats was significantly (P< or =0.05) greater than that of non-implant-side masseters; rats implanted with blank microsphere exhibited no significant difference between implant- and non-implant-side masseter COX activity levels. The stated null hypothesis was therefore rejected. These data suggest that TRH implants in proximity to trigeminal motoneurones effect increased oxidative capacity of the masseter muscle as measured by COX activity.

Biochemical properties of the superficial masseter muscle in the aging rat.

In young (15 weeks) and senescent (2 years) rats, the metabolic enzyme activities and the composition of myosin heavy chain (MHC) isoforms of the superficial masseter muscle were assessed by biochemical analysis. The succinate dehydrogenase activity and phosphofructokinase activity of the masseter muscle were significantly lower in senescent rats than in young rats. A large portion of the masseter muscle was composed of MHC IIb and IId isoforms in both young and senescent rats. As compared with young rats, no significant change in the composition of MHC isoforms was found in the masseter muscle of the senescent rats. These findings indicate that aging has a significant effect on the energy supply of the superficial masseter muscle.

Effects of 17 beta-oestradiol and 5 alpha-dihydrotestosterone on the expression of the muscle and heart types of lactate dehydrogenase isozymes in the masseter muscle of developing mice.

17 beta-oestradiol (E2) and/or 5 alpha-dihydrotestosterone (5 alpha-DHT) had no effect on the expression of isozymes of lactate dehydrogenase (LDH) in the masseter muscle of intact male mice. However, treatment with E2 restored the level of the muscle (M) type of LDH isozyme, which had been reduced by testectomy, to that found in intact male mice treated with vehicle only. Moreover, 5 alpha-DHT alone was more effective than E2 in increasing the relative level of this isozyme in testectomized mice. 5 alpha-DHT had a more significant effect on the increase in the relative level of the M-type LDH isozyme when combined with E2. These results suggest that androgens promote, in the presence of oestrogens, the postnatal changes in the characteristics of the masseter muscle of developing male animals.

A quantitative histochemical study of the spatial distribution of intrafascicular fibre types in the porcine masseter and soleus muscles.

The distribution of intrafascicular type I and II muscle fibres together with proportions of edge- and centrally located type I and II fibres within whole fascicles were analysed by myosin ATP-ase histochemistry in 241 porcine masseter fascicles (six masseter muscles) and compared with result from 63 pig soleus fascicles (five soleus muscles). All fascicles were from 11 domestic pigs (1 yr old, 70-90 kg body weight, all female). The proportions of type I fibres (slow) and type II fibres (fast) on the edge of fascicles differed significantly from the proportions centrally. All the soleus fascicles had higher proportions of centrally located type I fibres. Only seven out of 241 (3%) masseter fascicles diverged in this respect and showed reversed intrafascicular fibre-type proportions with more edge-located type I fibres. Analysis of the fascicular distribution of type I and II fibres revealed that the porcine masseter had type II fibres as the predominant type. Between 68-87% of the total fibres were type II (p < 0.001). The intrafascicular content of type I fibres increased towards the deep part of the masseter. In four of five soleus muscles the type II fibre population was dominant (p < 0.01). However, one soleus revealed equal proportions of type I and II fibres.