Adult Masseter Muscle Research Articles

Heterogeneity of adult masseter muscle satellite cells with cardiomyocyte differentiation potential

Although resident cardiac stem cells have been reported, regeneration of functional cardiomyocytes (CMs) remains a challenge. The present study identifies an alternative progenitor source for CM regeneration without the need for genetic manipulation or invasive heart biopsy procedures. Unlike limb skeletal muscles, masseter muscles (MM) in the mouse head are developed from Nkx2–5 mesodermal progenitors. Adult masseter muscle satellite cells (MMSCs) display heterogeneity in developmental origin and cell phenotypes. The heterogeneous MMSCs that can be characterized by cell sorting based on stem cell antigen-1 (Sca1) show different lineage potential. While cardiogenic potential is preserved in Sca1+ MMSCs as shown by expression of cardiac progenitor genes (including Nkx2–5), skeletal myogenic capacity is maintained in Sca1- MMSCs with Pax7 expression. Sca1+ MMSC-derived beating cells express cardiac genes and exhibit CM-like morphology. Electrophysiological properties of MMSC-derived CMs are demonstrated by calcium transients and action potentials. These findings show that MMSCs could serve as a novel cell source for cardiomyocyte replacement.

Molecular motor MYO1C, acetyltransferase KAT6B and osteogenetic transcription factor RUNX2 expression in human masseter muscle contributes to development of malocclusion

ObjectiveType I myosins are molecular motors necessary for glucose transport in the cytoplasm and initiation of transcription in the nucleus. Two of these, MYO1H and MYO1C, are paralogs which may be important in the development of malocclusion. The objective of this study was to investigate their gene expression in the masseter muscle of malocclusion subjects. Two functionally related proteins known to contribute to malocclusion were also investigated: KAT6B (a chromatin remodelling epigenetic enzyme which is activated by MYO1C) and RUNX2 (a transcription factor regulating osteogenesis which is activated by KAT6B). DesignMasseter muscle samples and malocclusion classifications were obtained from orthognathic surgery subjects. Muscle was sectioned and immunostained to determine fibre type properties. RNA was isolated from the remaining sample to determine expression levels for the four genes by TaqMan® RT-PCR. Fibre type properties, gene expression quantities and malocclusion classification were compared. ResultsThere were very significant associations (P<0.0000001) between MYO1C and KAT6B expressions. There were also significant associations (P<0.005) between RUNX2 expression and masseter muscle type II fibre properties. Very few significant associations were identified between MYO1C and masseter muscle fibre type properties. ConclusionsThe relationship between MYO1C and KAT6B suggests that the two are interacting in chromatin remodelling for gene expression. This is the nuclear myosin1 (NM1) function of MYO1C. A surprising finding is the relationship between RUNX2 and type II masseter muscle fibres, since RUNX2 expression in mature muscle was previously unknown. Further investigations are necessary to elucidate the role of RUNX2 in adult masseter muscle.

Intrafusal myosin heavy chain expression of human masseter and biceps muscles at young age shows fundamental similarities but also marked differences

Muscle spindles are skeletal muscle mechanoreceptors that provide proprioceptive information to the central nervous system. The human adult masseter muscle has greater number, larger and more complex muscle spindles than the adult biceps. For a better knowledge of muscle diversity and physiological properties, this study examined the myosin heavy chain (MyHC) expression of muscle spindle intrafusal fibres in the human young masseter and young biceps muscles by using a panel of monoclonal antibodies (mAbs) against different MyHC isoforms. Eight MyHC isoforms were detected in both muscles-slow-tonic, I, IIa, IIx, foetal, embryonic, α-cardiac and an isoform not previously reported in intrafusal fibres, termed IIx'. Individual fibres co-expressed 2-6 isoforms. MyHC-slow tonic separated bag1, AS-bag1 and bag2 fibres from chain fibres. Typically, bag fibres also expressed MyHC-I and α-cardiac, whereas chain fibres expressed IIa and foetal. In the young masseter 98% of bag1 showed MyHC-α cardiac versus 30% in the young biceps, 35% of bag2 showed MyHC-IIx' versus none in biceps, 17% of the chain fibres showed MyHC-I versus 61% in the biceps. In conclusion, the result showed fundamental similarities in intrafusal MyHC expression between young masseter and biceps, but also marked differences implying muscle-specific proprioceptive control, probably related to diverse evolutionary and developmental origins. Finding of similarities in MyHC expression between young and adult masseter and biceps muscle spindles, respectively, in accordance with previously reported similarities in mATPase fibre type composition suggest early maturation of muscle spindles, preceding extrafusal fibres in growth and maturation.

Human Masseter Muscle Fibers From the Elderly Express Less Neonatal Myosin Than Those of Young Adults

In contrast to limb muscles where neonatal myosin (MyHC-neo) is present only shortly after birth, adult masseter muscles contain a substantial portion of MyHC-neo, which is coexpressed with mature MyHC isoforms. Changes in the numerical and area proportion of muscle fibers containing MyHC-neo in masseter muscle with aging could be expected, based on previously reported findings that (i) developmental MyHC-containing muscle fibers exhibit lower shortening velocities compared to fibers with exclusively fast MyHC isoforms and (ii) transformation toward faster phenotype occurs in elderly compared to young masseter muscle. In this study, we detected MyHC isoforms in the anterior superficial part of the human masseter muscle in a sufficiently large sample of young, middle-aged, and elderly subjects to reveal age-related changes in the coexpression of MyHC-neo with adult MyHC isoforms. MyHC isoforms were visualized with immunoperoxidase method and the results were presented by (i) the area proportion of fibers containing particular MyHC isoforms and (ii) the numerical proportion of fiber types defined by MyHC-1, -2a, -2x, and -neonatal isoform expression from a successive transverse sections. We found a lower numerical and area proportion of fibers expressing MyHC-neo as well as a lower area proportion of fibers containing MyHC-1 in elderly than in young subjects. We conclude that the diminished expression of MyHC-neo with age could point to a lower regeneration capacity of masseter muscle in the elderly.

Expression of muscle-specific integrins in masseter muscle fibers during malocclusion disease

Integrins are heterodimeric cell surface membrane proteins linking the extracellular matrix to actin. α7B integrin is detected in proliferating and adult myofibers, whereas α7A plays a role in regenerating muscle fibers with a minor function in mature muscle fibers. The expression levels of β1A appear to be very low, whereas β1D appears to be the predominant integrin form in mature muscle. Considering the important features of masseter muscle we have studied integrin expression in masseter muscle specimens of surgical patients with posterior right crossbite and comparing them to left side masseter muscle specimens. Our results showed that the expression of integrins was significantly lower in the crossbite side muscle. Furthermore, the most important finding is that β1A is clearly detectable in adult masseter muscle. This behavior could be due to the particular composition of masseter, since it contains hybrid fibers showing the capacity to modify the contractile properties to optimize the energy efficiency or the action of the muscle during contraction. Moreover, masseter is characterized by a high turnover of muscle fibers producing a regeneration process. This may indicate a longer time to heal, justifying the loss of β1D and the consequential increase of β1A. Thus, our data provide the first suggestion that integrins in masseter muscle play a key role regulating the functional activity of muscle and allowing the optimization of contractile forces.

Characteristics of myofibres in the masseter muscle of mice during postnatal growth period.

Functional maturation of muscles is related to the constitutional proportion of muscle protein isoforms during development and growth. Although the mouse masseter muscle (MS) is classified as a fast limb muscle, its functions are different from those of a limb muscle. This study investigated the differentiation of myosin heavy chain (MHC) isoforms during the postnatal development periods in mouse MS and mouse tibialis anterior (TA), which is a fast limb muscle. Many anti-MHC slow-type-positive fibres were observed in neonatal MS and TA; these fibres decreased during development. Adult MS was composed of anti-MHC fast-type-positive fibres. MHC isoforms in MS were composed of MHC-2a and MHC-2d soon after birth. MHC-2b was expressed, but MHC-2a was not seen after 21 days. Expression of MHC-2b agreed with the weaning period, that is 2-3 weeks after birth. This fact suggested that the transformation from suckling to mastication changed the MHC isoforms during this period. In this study, the expressions of MHC-2b agree with the weaning period.

Effect of maturation on 31P magnetic resonance spectroscopy of the rabbit masseter muscle.

This work studies the dynamic metabolic changes of the rabbit masseter muscle during post-natal development. The composition and proportion of oxidative and glycolytic muscle fibers alter during maturation. The masseter muscle, as most muscles of the craniofacial region, exhibits unusual development in composition of isoforms of myosin. The effect of this unusual composition on the dynamic metabolic properties of the masseter muscle have not been assessed. The metabolism of the rabbit masseter muscle was studied by means of 31P-nuclear magnetic resonance (NMR) spectroscopy. Contraction was elicited by electrical stimulation of the muscle in the anesthetized animal. Five animals were studied at 8 weeks and 24 weeks so that both the juvenile and adult stages could be evaluated. The dynamic biochemical changes in the masseter muscle were studied by the analysis of NMR spectra. A single-turn surface coil (copper) was used, and the original signal was treated with Fourier transforms to obtain 31P spectra. The low signal-to-noise ratio required averaging 16 acquisitions (acquisition time = 400 msec, repetition rate = 1.8 sec) in 30 sec and then obtaining continuous spectra for 27 min. Each averaged spectrum demonstrated five peaks: inorganic phosphate (Pi), creatine phosphate (PCr), and three peaks related to adenosine triphosphate (ATP). The protocol involved recording an initial three-minute rest period, stimulating the muscle at 5 Hz for 3 min twice, separated by three-minute rest periods, and stimulating the muscle at 50 Hz twice for 3 min separated by rest periods. The Pi/PCr ratio increased significantly in the adult masseter during both 5-Hz stimulations, evoking twitching, and the first 50-Hz stimulation, evoking tetany (repeated ANOVA, P < 0.05). The resting pH (6.96 +/- 0.13) was significantly lowered during both twitching (6.85 +/- 0.10; P < 0.0038) and tetany (6.55 +/- 0.13; P < 0.0001), but only in the adult masseter muscle. These finding suggest that the adult masseter muscle possesses more glycolytic fibers as it modifies its metabolism during postnatal development.

The human embryonic myosin alkali light chain gene: use of alternative promoters and 3′ non-coding regions

Recently we have found evidence that the human embryonic myosin alkali light chain (MLC1 emb) gene has two functional promoters and that its mRNAs exhibit heterogeneity in their 3'untranslated regions (UTR). To study this more in detail we have isolated and characterized the human MLC1emb gene. We focussed in particular on 2 kilobases of 5'flanking region and the alternative 3'UTRs. RNA primer extension and S1 mapping analyses revealed that the MLC1emb gene can indeed be driven either by a proximal or a distal promoter, both in fetal and adult cardiac tissue. These MLC1emb RNAs can contain either the proximal or distal 3'UTR. In contrast to this, in fetal as well as adult masseter muscle MLC1emb mRNA is predominantly transcribed from the proximal promoter and contains mainly the distal 3'UTR. These results explain the known heterogeneity of MLC1emb mRNAs. Finally, we present evidence that the murine MLC1emb gene also contains a functional distal promoter element which has hitherto been undetected.