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Myosin Heavy Chain (myosin + heavy_chain)
Terms modified by Myosin Heavy Chain Selected AbstractsIncreased ,-Myosin Heavy Chain in Acute Cellular Rejection Following Human Heart TransplantationAMERICAN JOURNAL OF TRANSPLANTATION, Issue 4 2002Mohamad H. Yamani Background: Increased expression of smooth muscle and nonmuscle myosin heavy chains has been previously reported in animal models of cardiac allograft rejection. However, altered expression of ,-myosin heavy chain in human cardiac rejection has not been determined. Methods: Two-dimensional (2D)-gel electrophoresis of endomyocardial biopsies taken from patients with (Grade 3A, n = 6) and without (Grade 0, n = 6) acute rejection were analyzed. Increased expression of two protein spots (MW , 12 kDa) were identified in the presence of acute rejection and were further characterized by mass spectrometry analysis. In patients who had acute rejection, protein expression was subsequently analyzed by immunoblotting on biopsies preceding, during, and following treatment of rejection. Results: Mass spectrometric analysis of the protein spots detected 6 and 22 tryptic peptides, respectively. Protein sequence database search analysis identified the first protein as ,-myosin heavy chain and the second spot consisted of proteins of unidentified nature that may represent novel proteins. Immunoblotting analysis showed 1.4 × fold increase (p <,0.01) of protein expression of ,-myosin heavy chain expression in the presence of acute rejection. Conclusions: To our knowledge, this is the first 2D-gel study to describe increased expression of ,-myosin heavy chain and other proteins of unidentified nature in association with human cardiac allograft rejection. [source] Effects of Insulin-Like Growth Factor-1 Gene Transfer on Myosin Heavy Chains in Denervated Rat Laryngeal Muscle,THE LARYNGOSCOPE, Issue 2 2004Paul W. Flint MD Abstract Objectives/Hypothesis: To determine whether the myotrophic activity of human insulin-like growth factor (hIGF)-1 promotes restoration of normal myosin heavy chain (MHC) composition after nerve injury, MHC composition was analyzed after hIGF-1 gene transfer in denervated rat laryngeal muscle. Study Design: Animal model to study effects of gene transfer on laryngeal paralysis. Methods: In anesthetized rats, the left recurrent and superior laryngeal nerves are cut and suture ligated. A midline thyrotomy is performed, and the thyroarytenoid muscle is injected with a polyvinyl-based formulation containing a muscle specific expression system and hIGF-1 DNA (treatment group) or saline (control group). After 30 days, animals were killed, and the thyroarytenoid muscle was removed and processed for sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDSPAGE). Densitometric measurements were obtained to determine composition of MHCs. Results: As previously described, MHC composition in denervated laryngeal muscle was characterized by a decrease in type IIB and IIL and up-regulation of IIA/IIX. Compared with controls, hIGF-1 treated animals demonstrated a significant increase in expression of type IIB and IIL and a significant decrease in expression of type IIA/X. Conclusions: These findings suggest that the myotrophic effect of hIGF-1 gene transfer results in normalization of MHC composition in denervated muscle, with suppression of type IIA/X MHC and promotion of type IIL expression. [source] Clenbuterol antagonizes glucocorticoid-induced atrophy and fibre type transformation in miceEXPERIMENTAL PHYSIOLOGY, Issue 1 2004Maria Antonietta Pellegrino Beta-agonists and glucocorticoids are frequently coprescribed for chronic asthma treatment. In this study the effects of 4 week treatment with beta-agonist clenbuterol (CL) and glucocorticoid dexamethasone (DEX) on respiratory (diaphragm and parasternal) and limb (soleus and tibialis) muscles of the mouse were studied. Myosin heavy chain (MHC) distribution, fibres cross sectional area (CSA), glycolytic (phosphofructokinase, PFK; lactate dehydrogenase, LDH) and oxidative enzyme (citrate synthase, CS; cytochrome oxidase, COX) activities were determined. Muscle samples were obtained from four groups of adult C57/B16 mice: (1) Control (2) Mice receiving CL (CL, 1.5 mg kg,1 day,1 in drinking water) (3) Mice receiving DEX (DEX, 5.7 mg kg,1 day,1s.c.) (4) Mice receiving both treatments (DEX + CL). As a general rule, CL and DEX showed opposite effects on CSA, MHC distribution, glycolytic and mitochondrial enzyme activities: CL alone stimulated a slow-to-fast transition of MHCs, an increase of PFK and LDH and an increase of muscle weight and fibre CSA; DEX produced an opposite (fast-to-slow transition) change of MHC distribution, a decrease of muscle weight and fibre CSA and in some case an increase of CS. The response varied from muscle to muscle with mixed muscles, as soleus and diaphragm, being more responsive than fast muscles, as tibialis and parasternal. In combined treatments (DEX + CL), the changes induced by DEX or CL alone were generally minimized: in soleus, however, the effects of CL predominated over those of DEX, whereas in diaphragm DEX prevailed over CL. Taken together the results suggest that CL might counteract the unwanted effects on skeletal muscles of chronic treatment with glucocorticoids. [source] Regional specialization of rat quadriceps myosin heavy chain isoforms occurring in distal to proximal parts of middle and deep regions is not mirrored by citrate synthase activityJOURNAL OF ANATOMY, Issue 1 2007Tertius Abraham Kohn Abstract Myosin heavy chain (MHC) isoform content and citrate synthase (CS) activities were measured in the Quadriceps femoris (QF) muscle of 18 female rats. The muscle group was divided into superficial, middle and deep, distal, central and proximal parts. MHC IIb and IIx were more abundant in superficial regions (P < 0.05) with low CS activities compared with deeper parts. The deeper parts expressed all four isoforms (MHC IIb, MHC IIx, MHC IIa and MHC I), with a concomitantly higher CS activity. MHC I, MHC IIa and MHC IIb isoform content varied significantly along the length of the deep regions. Only MHC IIb and CS activity in the proximal middle part correlated (negatively) with each other. This study showed that the QF has regional specialization and that standardization of sampling site is important. Furthermore, CS activity and MHC isoforms are only loosely associated, or not at all. [source] IDENTIFICATION OF A MYOFIBRIL-BOUND SERINE PROTEINASE IN THE SKELETAL MUSCLE OF SILVER CARPJOURNAL OF FOOD BIOCHEMISTRY, Issue 5 2004MIN-JIE CAO ABSTRACT Myofibril-bound serine proteinase (MBSP) in the skeletal muscle of silver carp was characterized. Myosin heavy chain (MHC) degraded markedly when silver carp myofibril was incubated at 55,60C as shown by SDS-PAGE. Prolonged incubation of myofibrils also caused the degradation of other myofibrillar proteins such as ,-actinin, actin and tropomyosin to some degree. The results suggest the existence of an endogenous myofibril associated proteinase. Serine proteinase inhibitors (Pefabloc SC and Lima bean trypsin inhibitor) greatly suppressed the degradation of myosin heavy chain, while inhibitors for cysteine, metallo, and aspartic proteinases did not show any effect, indicating that the endogeneous proteinase is a myofibril-bound serine proteinase. [source] Developmental expression and comparative genomic analysis of Xenopus cardiac myosin heavy chain genesDEVELOPMENTAL DYNAMICS, Issue 4 2005Robert J. Garriock Abstract Myosin heavy chains (MHC) are cytoskeletal motor proteins essential to the process of muscle contraction. We have determined the complete sequences of the Xenopus cardiac MHC genes, ,-MHC and ventricular MHC (vMHC), and have characterized their developmental expression profiles. Whereas ,-MHC is expressed from the earliest stages of cardiac differentiation, vMHC transcripts are not detected until the heart has undergone chamber formation. Early expression of vMHC appears to mark the cardiac conduction system, but expression expands to include the ventricle and outflow tract myocardium during subsequent development. Sequence comparisons, transgenic expression analysis, and comparative genomic studies indicate that Xenopus ,-MHC is the true orthologue of the mammalian ,-MHC gene. On the other hand, we show that the Xenopus vMHC gene is most closely related to chicken ventricular MHC (vMHC1) not the mammalian ,-MHC. Comparative genomic analysis has allowed the detection of a mammalian MHC gene (MyH15) that appears to be the orthologue of vMHC, but evidence suggests that this gene is no longer active. Developmental Dynamics 233:1287,1293, 2005. © 2005 Wiley-Liss, Inc. [source] Protective effects of exercise preconditioning on hindlimb unloading-induced atrophy of rat soleus muscleACTA PHYSIOLOGICA, Issue 1 2009H. Fujino Abstract Aim:, A chronic decrease in the activation and loading levels of skeletal muscles as occurs with hindlimb unloading (HU) results in a number of detrimental changes. Several proteolytic pathways are involved with an increase in myofibrillar protein degradation associated with HU. Exercise can be used to counter this increase in proteolytic activity and, thus, may be able to protect against some of the detrimental changes associated with chronic decreased use. The purpose of the present study was to determine the potential of a single bout of preconditioning endurance exercise in attenuating the effects of 2 weeks of HU on the mass, phenotype and force-related properties of the soleus muscle in adult rats. Methods:, Male Wistar rats were subjected to HU for 2 weeks. One half of the rats performed a single bout of treadmill exercise for 25 min immediately prior to the 2 weeks of HU. Results:, Soleus mass, maximum tetanic tension, myofibrillar protein content, fatigue resistance and percentage of type I (slow) myosin heavy chain were decreased in HU rats. In addition, markers for the cathepsin, calpain, caspase and ATP-ubiquitin-proteasome proteolytic pathways were increased. The preconditioning endurance exercise bout attenuated all of the detrimental changes associated with HU, and increased HSP72 mRNA expression and protein levels. Conclusion:, These findings indicate that exercise preconditioning may be an effective countermeasure to the detrimental effects of chronic decreases in activation and loading levels on skeletal muscles and that an elevation in HSP72 may be one of the mechanisms associated with these responses. [source] Different adaptations of alpha-actinin isoforms to exercise training in rat skeletal musclesACTA PHYSIOLOGICA, Issue 3 2009Y. Ogura Abstract Aim:, Alpha (,)-actinins are located in the skeletal muscle Z-line and form actin,actin cross-links. Mammalian skeletal muscle has two isoforms: ,-actinin-2 and ,-actinin-3. However, the response of ,-actinin to exercise training is little understood. Therefore, the current study examined the effects of exercise training on the expression level of two ,-actinin isoforms in skeletal muscles. Methods:, Twelve male Wistar rats were assigned randomly to a control (C; n = 6) or exercise training (T; n = 6) group. After T animals were trained on an animal treadmill for 9 weeks, ,-actinin-2 and ,-actinin-3 levels in the plantaris, white and red gastrocnemius muscles were analysed. In addition, changes in the myosin heavy chain (MyHC) composition were assessed, and muscle bioenergetic enzyme activities were measured. Results:, Results show that exercise training increased ,-actinin-2 expression levels in all muscles (P < 0.05). However, no significant difference was found in ,-actinin-3 expression levels between C and T animals. Subsequent MyHC analyses of all muscle showed an MyHC shift with direction from IIb to IIa. Furthermore, enzymatic analysis revealed that exercise training improved enzyme activities related to aerobic metabolism. Conclusion:, The results of this study demonstrate that exercise training alters the expression level of ,-actinin at the isoform level. Moreover, the increase in expression levels of ,-actinin-2 is apparently related to alteration of skeletal muscle: its aerobic capacity is improved. [source] Androgen replacement therapy improves function in male rat muscles independently of hypertrophy and activation of the Akt/mTOR pathwayACTA PHYSIOLOGICA, Issue 4 2009C. Hourdé Abstract Aim:, We analysed the effect of physiological doses of androgens following orchidectomy on skeletal muscle and bone of male rats, as well as the relationships between muscle performance, hypertrophy and the Akt/mammalian target of rapamycin (mTOR) signalling pathway involved in the control of anabolic and catabolic muscle metabolism. Methods:, We studied the soleus muscle and tibia from intact rats (SHAM), orchidectomized rats treated for 3 months with vehicle (ORX), nandrolone decanoate (NAN) or dihydrotestosterone (DHT). Results:, Orchidectomy had very little effect on the soleus muscle. However, maximal force production by soleus muscle (+69%) and fatigue resistance (+35%) in NAN rats were both increased when compared with ORX rats. In contrast, DHT treatment did not improve muscle function. The relative number of muscle fibres expressing slow myosin heavy chain and citrate synthase activity were not different in NAN and ORX rats. Moreover, NAN and DHT treatments did not modify muscle weights and cross-sectional area of muscle fibres. Furthermore, phosphorylation levels of downstream targets of the Akt/mTOR signalling pathway, Akt, ribosomal protein S6 and eukaryotic initiation factor 4E-binding protein 1 were similar in muscles of NAN, DHT and ORX rats. In addition, trabecular tibia from NAN and DHT rats displayed higher bone mineral density and bone volume when compared with ORX rats. Only in NAN rats was this associated with increased bone resistance to fracture. Conclusion:, Physiological doses of androgens are beneficial to muscle performance in orchidectomized rats without relationship to muscle and fibre hypertrophy and activation of the Akt/mTOR signalling pathway. Taken together our data clearly indicate that the activity of androgens on muscle and bone could participate in the global improvement of musculoskeletal status in the context of androgen deprivation induced by ageing. [source] Correlation of dystrophin,glycoprotein complex and focal adhesion complex with myosin heavy chain isoforms in rat skeletal muscleACTA PHYSIOLOGICA, Issue 4 2009S. Masuda Abstract Aim:, The dystrophin,glycoprotein complex (DGC) and focal adhesion complex (FAC) are transmembrane structures in muscle fibres that link the intracellular cytoskeleton to the extracellular matrix. DGC and FAC proteins are abundant in slow-type muscles, indicating the structural reinforcement which play a pivotal role in continuous force output to maintain posture for long periods. The aim of the present study was to examine the expression of these structures across fast-type muscles containing different myosin heavy chain (MHC) isoform patterns which reflect the fatigue-resistant characteristics of skeletal muscle. Methods:, We measured the expression of dystrophin and ,1 integrin (representative proteins of DGC and FAC respectively) in plantaris, extensor digitorum longus, tibialis anterior, red and white portions of gastrocnemius, superficial portion of vastus lateralis and diaphragm, in comparison with soleus (SOL) and cardiac muscle from rats. Results:, The expression of dystrophin and ,1 integrin correlated positively with the percentage of type I, IIa and IIx MHC isoforms and negatively with that of type IIb MHC isoform in fast-type skeletal muscles, and their expression was abundant in SOL and cardiac muscle. Conclusion:, Our results support the idea that DGC and FAC are among the factors that explain the fatigue-resistant property not only of slow-type but also of fast-type skeletal muscles. [source] In vivo phosphorylation of regulatory light chain of myosin II in sea urchin eggs and its role in controlling myosin localization and function during cytokinesisCYTOSKELETON, Issue 2 2008Ryota Uehara Abstract Phosphorylation of myosin regulatory light chain (RLC) at Ser19 (mono-phosphorylation) promotes filament assembly and enhances actin-activated ATPase activity of non-muscle myosin, while phosphorylation at both Ser19 and Thr18 (di-phosphorylation) further enhances the ATPase activity. However, it has not well been addressed which type of phosphorylation is important in regulating myosin during cytokinesis. Here, we investigated subcellular localization in sea urchin eggs of mono-phosphorylated and di-phosphorylated RLC by both quantitative biochemical and spatiotemporal cytological approaches. Mono-phosphorylated RLC was dominant in the equatorial cortex throughout the whole process of cytokinesis. Inhibition of myosin light chain kinase (MLCK) decreased mono-phosphorylated RLC both in the cortex and in the cleavage furrow, and blocked both formation and contraction of the contractile ring. Two different types of ROCK inhibitor gave inconsistent results: H1152 blocked both RLC mono-phosphorylation in the cleavage furrow and contraction of the contractile ring, while Y27632 affected neither the mono-phosphorylation nor cell division. These results suggest that there may be other targets of H1152 than ROCK, which is involved in the RLC phosphorylation in the cleavage furrow. Furthermore, it was revealed that localization of myosin heavy chain in the cleavage furrow, but not in the cortex, was perturbed by inhibition of RLC mono-phosphorylation. These results suggested that RLC mono-phosphorylation by more than two RLC kinases play a main role in regulation and localization of myosin in the dividing sea urchin eggs. Cell Motil. Cytoskeleton 2007. © 2007 Wiley-Liss, Inc. [source] Promoter analysis of ventricular myosin heavy chain (vmhc) in zebrafish embryosDEVELOPMENTAL DYNAMICS, Issue 7 2009Daqing Jin Abstract In zebrafish, ventricular myosin heavy chain (vmhc) gene is initially expressed at the anterior lateral mesoderm and thereafter its expression is restricted to the cardiac ventricle. The transcriptional control mechanisms in regulating chamber-specific expression of myosin heavy chains are not well defined. We isolated and analyzed zebrafish vmhc upstream region to examine the spatial and temporal regulation of vmhc using transgenic and transient expression techniques. Promoter deletion analyses defined a basal promoter region sufficient to drive vmhc expression in the ventricle and an upstream fragment necessary for repressing ectopic vmhc expression in the atrium. The transcriptional mechanism that prevents vmhc expression in the atrium is mediated through Nkx2.5 binding elements (NKE). We have further discovered that paired-related homeobox transcriptional factor 2 (Prx2/S8)-like binding elements are required for promoting vmhc expression, and Prrx1b, a Prx-related homeobox protein, participates in the regulation of vmhc expression with other transcriptional factors. Developmental Dynamics 238:1760,1767, 2009. © 2009 Wiley-Liss, Inc. [source] Electrophoretic variants of cardiac myosin heavy chain-, in Sprague Dawley ratsELECTROPHORESIS, Issue 3 2004Peter J. Reiser Abstract Analysis of cardiac myosin revealed differences in gel electrophoretic migration patterns of the ,-isoform of myosin heavy chain, but not the ,-isoform, in Sprague Dawley rats. No differences in the migration patterns of the ,-or ,-isoforms were observed in other rat strains. Three electrophoretic migration patterns of the ,-isoforms were observed in individual rats: a slower migrating isoform alone (4% of all rats tested), a faster migrating isoform alone (55%), and both isoforms (41%). The isoform expression pattern was identical in all myocardial regions in each rat. Frequency of expression patterns suggests multiple gene sequences for ,-cardiac myosin heavy chain in Sprague Dawley rats. Sequence analysis of amplified regions of the Sprague Dawley and Brown Norway rat ,-myosin genes, specifically the 5'-untranslated region, exons 1,3, and associated introns, showed numerous single nucleotide polymorphisms in coding and noncoding regions, including putative regulatory sites in Sprague Dawley rats, but not in Brown Norway rats. All Sprague Dawley rats varied from Brown Norway rats and no heterogeneity was observed in Brown Norway rats. Several deletions and dimorphic positions were also observed. Dimorphic positions were evident on automated sequencing comparisons. The data indicate that at least two ,-myosin heavy chain isoforms exist in Sprague Dawley rats and these rats exhibit sequence diversity within that portion of the ,-myosin heavy chain gene reported in this study. [source] Vertical agarose gel electrophoresis and electroblotting of high-molecular-weight proteinsELECTROPHORESIS, Issue 11 2003Chad M. Warren Abstract The electrophoretic separation of high-molecular-weight proteins (>,500 kDa) using polyacrylamide is difficult because gels with a large enough pore size for adequate protein mobility are mechanically unstable. A 1% vertical sodium dodecyl sulfate (SDS)-agarose gel electrophoresis (VAGE) system has been developed that allows titin (a protein with the largest known SDS subunit size of 3000,4000 kDa) to migrate over 10 cm in a ,13 cm resolving gel. Such migration gives clear and reproducible separation of titin isoforms. Proteins ranging in size from myosin heavy chain (,,220 kDa) up to titin can be resolved on this gel system. Electroblotting of these very large proteins was nearly 100% efficient. This VAGE system has revealed two titin size variants in rabbit psoas muscle, two N2BA bands in rabbit cardiac muscle, and species differences between titins from rat and rabbit muscle. Agarose electrophoresis should be the method of choice for separation and blotting of proteins with very large subunit sizes. [source] Effect of neurotrophin-3 on reinnervation of the larynx using the phrenic nerve transfer techniqueEUROPEAN JOURNAL OF NEUROSCIENCE, Issue 2 2007Paul J. Kingham Abstract Current techniques for reinnervation of the larynx following recurrent laryngeal nerve (RLN) injury are limited by synkinesis, which prevents functional recovery. Treatment with neurotrophins (NT) may enhance nerve regeneration and encourage more accurate reinnervation. This study presents the results of using the phrenic nerve transfer method, combined with NT-3 treatment, to selectively reinnervate the posterior cricoarytenoid (PCA) abductor muscle in a pig nerve injury model. RLN transection altered the phenotype and morphology of laryngeal muscles. In both the PCA and thyroarytenoid (TA) adductor muscle, fast type myosin heavy chain (MyHC) protein was decreased while slow type MyHC was increased. These changes were accompanied with a significant reduction in muscle fibre diameter. Following nerve repair there was a progressive normalization of MyHC phenotype and increased muscle fibre diameter in the PCA but not the TA muscle. This correlated with enhanced abductor function indicating the phrenic nerve accurately reinnervated the PCA muscle. Treatment with NT-3 significantly enhanced phrenic nerve regeneration but led to only a small increase in the number of reinnervated PCA muscle fibres and minimal effect on abductor muscle phenotype and morphology. Therefore, work exploring other growth factors, either alone or in combination with NT-3, is required. [source] Expression of muscle-related genes and two MyoD genes during amphioxus notochord developmentEVOLUTION AND DEVELOPMENT, Issue 5 2003Aki Urano Summary The notochord is one of the diagnostic features of the phylum Chordata. Despite the similarities in the early morphogenetic patterns of the notochords of various chordates, they are strikingly distinct from one another at the histological level. The amphioxus notochord is one example of an evolutionary novelty because it is made up of muscle cells. Our previous expressed sequence tag analysis, targeting messenger RNAs expressed in the adult amphioxus notochord, demonstrated that many muscle-related genes are expressed there. To characterize amphioxus notochord cells and to gain insights into the myogenic program in the notochord, we determined the spatial and temporal expre-ssion patterns of these muscle-related genes during amphioxus development. We found that BbNA1 (notochord actin), Amphi-Trop I (troponin I), Amphi-TPmyosin (tropo-myosin), Amphi-MHC2 (myosin heavy chain), Amphi-nMRLC (notochord-specific myosin regulatory light chain), Amphi-nTitin/MLCK (notochord-specific titin/myosin light chain kinase), Amphi-MLP/CRP3 (muscle LIM protein), and Amphi-nCalponin (notochord-specific calponin) are expres-sed with characteristic patterns in notochord cells, including the central cells, dorsally located cells, and ventrally located cells, suggesting that each notochord cell has a unique molecular architecture that may reflect its function. In addition, we characterized two MyoD genes (Amphi-MyoD1 and Amphi-MyoD2) to gain insight into the genetic circuitry governing the formation of the notochord muscle. One of the MyoD genes (Amphi-MyoD2) is expressed in the central notochord cells, and the coexistence of Amphi-MyoD2 transcripts along with the Amphi-MLP/CRP3 transcripts implies the participation of Amphi-MyoD2 in the myogenic program in the notochord muscle. [source] Plasticity of human skeletal muscle: gene expression to in vivo functionEXPERIMENTAL PHYSIOLOGY, Issue 5 2007Stephen D. R. Harridge Human skeletal muscle is a highly heterogeneous tissue, able to adapt to the different challenges that may be placed upon it. When overloaded, a muscle adapts by increasing its size and strength through satellite-cell-mediated mechanisms, whereby protein synthesis is increased and new nuclei are added to maintain the myonuclear domain. This process is regulated by an array of mechanical, hormonal and nutritional signals. Growth factors, such as insulin-like growth factor I (IGF-I) and testosterone, are potent anabolic agents, whilst myostatin acts as a negative regulator of muscle mass. Insulin-like growth factor I is unique in being able to stimulate both the proliferation and the differentiation of satellite cells and works as part of an important local repair and adaptive mechanism. Speed of movement, as characterized by maximal velocity of shortening (Vmax), is regulated primarily by the isoform of myosin heavy chain (MHC) contained within a muscle fibre. Human fibres can express three MHCs: MHC-I, -IIa and -IIx, in order of increasing Vmax and maximal power output. Training studies suggest that there is a subtle interplay between the MHC-IIa and -IIx isoforms, with the latter being downregulated by activity and upregulated by inactivity. However, switching between the two main isoforms appears to require significant challenges to a muscle. Upregulation of fast gene programs is caused by prolonged disuse, whilst upregulation of slow gene programs appears to require significant and prolonged activity. The potential mechanisms by which alterations in muscle composition are mediated are discussed. The implications in terms of contractile function of altering muscle phenotype are discussed from the single fibre to the whole muscle level. [source] Effect of anti-inflammatory and antioxidant drugs on the long-term repair of severely injured mouse skeletal muscleEXPERIMENTAL PHYSIOLOGY, Issue 4 2005A. Vignaud Non-steroidal anti-inflammatory drugs are frequently prescribed after skeletal muscle injury. It is not known whether this type of medication can interfere with muscle repair, although inflammatory response is thought to play an important role in this process. Tibialis anterior muscles of mice were injured by myotoxic agent (snake venom) or crushed. Then, animals were treated daily for 10,14 days with different types of non-steroidal anti-inflammatory and antioxidant drugs. The long-term repair was studied 10,42 days after injury by analysing the recovery of in situ muscle force production, size of regenerating muscle cells and expression of myosin heavy chain. Our results show that diclofenac, diferuloylmethane (curcumin), dimethylthiourea or pyrrolidine dithiocarbamate treatment did not significantly affect muscle recovery after myotoxic injury (P > 0.05). Similarly, diferuloylmethane, dimethyl sulphoxide or indomethacin administration did not markedly change muscle repair after crush injury. However, we noted that high doses (> 2 mg kg,1) of diferuloylmethane or indomethacin increased lethality and reduced muscle repair after crush injury. In conclusion, non-steroidal anti-inflammatory and antioxidant drugs did not exhibit long-term detrimental effects on muscle recovery after injury, except at lethal doses. [source] Functional regions in the essential light chain of smooth muscle myosin as revealed by the mutagenesis approachFEBS JOURNAL, Issue 20 2000Sophie Quevillon-Chéruel The endogenous essential light chain (LC17) of myosin from intestine smooth muscle was replaced with mutated essential light chains prepared using recombinant techniques. Complete exchange was observed with histidine-tagged derivatives of LC17a, LC17b and E122A-LC17a (LC17a and LC17b are the usual constituants of smooth muscle myosin), with small changes in the ATPase activity of reconstituted myosins. Much less exchange was observed with the light-chain derivative lacking the last 12 amino acid residues, demonstrating the importance of this segment, which may act as one arm of a pair of pincers to bind the myosin heavy chain. [source] Non-muscle myosin heavy chain (MYH9): A new partner fused to ALK in anaplastic large cell lymphomaGENES, CHROMOSOMES AND CANCER, Issue 4 2003Laurence Lamant In anaplastic large cell lymphoma, the ALK gene at 2p23 is known to be fused to NPM, TPM3, TPM4, TFG, ATIC, CLTC, MSN, and ALO17. All of these translocations result in the expression of chimeric ALK transcripts that are translated into fusion proteins with tyrosine kinase activity and oncogenic properties. We report a case showing a restricted cytoplasmic staining pattern of ALK and a novel chromosomal abnormality, t(2;22)(p23;q11.2), demonstrated by fluorescence in situ hybridization analysis. The result of 5, RACE analysis showed that the ALK gene was fused in-frame to a portion of the non-muscle myosin heavy chain gene, MYH9. Nucleotide sequence of the MYH9-ALK chimeric cDNA revealed that the ALK breakpoint was different from all those previously reported. It is localized in the same exonic sequence as MSN-ALK, but 6 bp downstream, resulting in an in-frame fusion of the two partner proteins. In contrast to the previously reported ALK fusion proteins, MYH9-ALK may lack a functional oligomerization domain. However, biochemical analysis showed that the new fusion protein is tyrosine phosphorylated in vivo but seems to lack tyrosine kinase activity in vitro. If further investigations confirm this latter result, the in vivo tyrosine phosphorylation of MYH9-ALK protein could involve mechanisms different from those described in the other ALK hybrid proteins. © 2003 Wiley-Liss, Inc. [source] The suitability of muscle of Cirrhinus mrigala in the formation of gel: a comparative electrophoretic study of six tropical carp meatsINTERNATIONAL JOURNAL OF FOOD SCIENCE & TECHNOLOGY, Issue 1 2008Rupsankar Chakrabarti Summary The annual inland fish production in India is 3.2 million tonnes and above. Aquaculture is around 80% of total inland fish production. The enhanced aquaculture production demands alternate processing methods for better utilisation of the farmed fish. Cirrhinus mrigala is one of the dominated species among cultured Indian major carps, but rated with lowest price. Fish meat with good gel-forming capacity is a prerequisite in the production of fast-moving fabricated analogue product. Cirrhinus mrigala had the highest gel strength, i.e. 435 gcm of its fresh meat in comparison with other carps. Apart from the highest protein and salt-soluble protein nitrogen content in fresh C. mrigala meat, this article also reports that the presence of the darkest thick band of myosin heavy chain and actin in the polyacrylamide gel electrophoresis pattern of salt-soluble extract of C. mrigala might be another possible reason for the highest gel strength in comparison with other carps. Cirrhinus mrigala contained white meat around 90%. This carp could be suitably used in the preparation of high-valued fabricated analogue product. [source] Patterning of the axial musculature in the developing chick embryo (Gallus)JOURNAL OF ANATOMY, Issue 5 2002P. J. Adds While the development and patterning events of the skeletal, myogenic and connective tissues of the developing limb buds of the chick have been relatively well studied, there is little known about the formation of the epaxial muscles and tendons. The epaxial muscles form the postvertebral muscle groups and develop from the myotome of the somite. The myotome develops from myogenic precursors migrating from the dorsomedial lip of the dermomyotome. These myogenic cells differentiate in a cranial to caudal sequence with the muscle fibres also orientated in a cranial to caudal direction. Here we use immunohistological staining both in whole mount and in transverse sections of chick embryos from various stages of development. Antibodies to the myosin heavy chain or tenascin were used to visualise the development of the epaxial muscles and tendons. The first myotomal muscle fibres are differentiated by Hamburger and Hamilton (H & H) stage 10 in the cranial somites, and differentiation proceeds caudally until at least H & H stage 22,23. Further development of the epaxial muscles does not take place until H & H stage 26,27 with the splitting of the myotome into the individual muscles. We demonstrate how the myotome splits into the individual muscles and how some muscle fibres become reorientated into a more oblique orientation. This delayed development and reorientation of muscle fibres is unique to the epaxial muscles. [source] Close relation of arterial ICC-like cells to the contractile phenotype of vascular smooth muscle cellJOURNAL OF CELLULAR AND MOLECULAR MEDICINE, Issue 4 2007Vladimír Pucovský Abstract This work aimed to establish the lineage of cells similar to the interstitial cells of Cajal (ICC), the arterial ICC-like (AIL) cells, which have recently been described in resistance arteries, and to study their location in the artery wall. Segments of guinea-pig mesenteric arteries and single AIL cells freshly isolated from them were used. Confocal imaging of immunostained cells or segments and electron microscopy of artery segments were used to test for the presence and cellular localization of selected markers, and to localize AIL cells in intact artery segments. AIL cells were negative for PGP9.5, a neural marker, and for von Willebrand factor (vWF), an endothelial cell marker. They were positive for smooth muscle ,-actin and smooth muscle myosin heavy chain (SM-MHC), but expressed only a small amount of smoothelin, a marker of contractile smooth muscle cells (SMC), and of myosin light chain kinase (MLCK), a critical enzyme in the regulation of smooth muscle contraction. Cell isolation in the presence of latrunculin B, an actin polymerization inhibitor, did not cause the disappearance of AIL cells from cell suspension. The fluorescence of basal lamina protein collagen IV was comparable between the AIL cells and the vascular SMCs and the fluorescence of laminin was higher in AIL cells compared to vascular SMCs. Moreover, cells with thin processes were found in the tunica media of small resistance arteries using transmis-sion electron microscopy. The results suggest that AIL cells are immature or phenotypically modulated vascular SMCs constitutively present in resistance arteries. [source] Effects of aging and gender on the spatial organization of nuclei in single human skeletal muscle cellsAGING CELL, Issue 5 2010Alexander Cristea Summary The skeletal muscle fibre is a syncitium where each myonucleus regulates the gene products in a finite volume of the cytoplasm, i.e., the myonuclear domain (MND). We analysed aging- and gender-related effects on myonuclei organization and the MND size in single muscle fibres from six young (21,31 years) and nine old men (72,96 years), and from six young (24,32 years) and nine old women (65,96 years), using a novel image analysis algorithm applied to confocal images. Muscle fibres were classified according to myosin heavy chain (MyHC) isoform expression. Our image analysis algorithm was effective in determining the spatial organization of myonuclei and the distribution of individual MNDs along the single fibre segments. Significant linear relations were observed between MND size and fibre size, irrespective age, gender and MyHC isoform expression. The spatial organization of individual myonuclei, calculated as the distribution of nearest neighbour distances in 3D, and MND size were affected in old age, but changes were dependent on MyHC isoform expression. In type I muscle fibres, average NN-values were lower and showed an increased variability in old age, reflecting an aggregation of myonuclei in old age. Average MND size did not change in old age, but there was an increased MND size variability. In type IIa fibres, average NN-values and MND sizes were lower in old age, reflecting the smaller size of these muscle fibres in old age. It is suggested that these changes have a significant impact on protein synthesis and degradation during the aging process. [source] IDENTIFICATION OF A MYOFIBRIL-BOUND SERINE PROTEINASE IN THE SKELETAL MUSCLE OF SILVER CARPJOURNAL OF FOOD BIOCHEMISTRY, Issue 5 2004MIN-JIE CAO ABSTRACT Myofibril-bound serine proteinase (MBSP) in the skeletal muscle of silver carp was characterized. Myosin heavy chain (MHC) degraded markedly when silver carp myofibril was incubated at 55,60C as shown by SDS-PAGE. Prolonged incubation of myofibrils also caused the degradation of other myofibrillar proteins such as ,-actinin, actin and tropomyosin to some degree. The results suggest the existence of an endogenous myofibril associated proteinase. Serine proteinase inhibitors (Pefabloc SC and Lima bean trypsin inhibitor) greatly suppressed the degradation of myosin heavy chain, while inhibitors for cysteine, metallo, and aspartic proteinases did not show any effect, indicating that the endogeneous proteinase is a myofibril-bound serine proteinase. [source] ATPASE ACTIVITY, SURFACE HYDROPHOBICITY, SULFHYDRYL CONTENT AND PROTEIN DEGRADATION IN REFRIGERATED SEABASS MUSCLE IN MODIFIED ATMOSPHERE PACKAGINGJOURNAL OF FOOD BIOCHEMISTRY, Issue 1 2004PAYAP MASNIYOM The effect of modified atmosphere packaging (80% CO2, 10% O2, 10% N2) on ATPase activity, surface hydrophobicity, sulfhydryl content and degradation of proteins in seabass muscle during storage at 4C was investigated. No changes in Ca2+ -, Mg2+ -, Mg2+ -Ca2+ -ATPase activities of natural actomyosin (NAM) in seabass slices kept under MAP were observed throughout the storage for up to 21 days (P > 0.05). However, a slightly increased Mg2+ -EGTA-ATPase was found. For seabass slices stored under air atmosphere, Ca2+ -ATPase activity decreased, whereas Mg2+ -EGTA-ATPase activity increased (P < 0.05) with a concomitant loss in Ca2+ -sensitivity. Lower decreases in total sulfhydryl content but higher increases in surface hydrophobicity were observed in samples stored under MAP, compared to those kept under air atmosphere. No marked autolytic degradation in samples kept under MAP was observed throughout the storage as monitored by no changes in myosin heavy chain, free ,-amino acid and trichloroacetic acid soluble peptide. Conversely, a considerable degradation was found in samples kept under air atmosphere, especially after 9 days of storage. Therefore, MAP is a promising means to retard the changes in muscle proteins, especially degradation. [source] DIFFERENT ARRANGEMENT OF ,-(,-GLUTAMYL)LYSINE CROSS-LINKING IN ALASKA POLLOCK (THERAGRA CHALCOGRAMMA) SURIMI PROTEINS BY STREPTOVERTICILLIUM AND ENDOGENOUS TRANSGLUTAMINASES DURING SUWARI PROCESSJOURNAL OF FOOD BIOCHEMISTRY, Issue 5 2001KENJI SATO ABSTRACT The objective of the present study is to compare the protein cross-linking reaction in Alaska pollock surimi that is catalyzed by a commercially available microbial transglutaminase and by endogenous Alaska pollock transglutaminase. The endogenous transglutaminase was inhibited by EGTA and activated by CaCl2 The microbial transglutaminase was added to the salted surimi with and without EGTA and CaCl2. These surimi pastes were incubated at 25C up to 24 h followed by cooking at 90C. The resultant gels were fractionated into soluble and insoluble (aggregate) fractions by SDS-urea extraction. Compositional analysis revealed that the aggregate consisted predominantly of cross-linked myosin heavy chain. The distribution of ,-(,-glutamyl)lysine isopeptide in the soluble and aggregate fractions andpeptide mapping analyses of the aggregate fraction demonstrate that the formation of isopeptide cross-links in Alaska pollock surimi proteins during suwari process differs when catalyzed by the microbial transglutaminase and endogenous transglutaminase. [source] PORCINE PLASMA PROTEINS AS GEL ENHANCER IN BIGEYE SNAPPER (PRIACANTHUS TAYENUS) SURIMIJOURNAL OF FOOD BIOCHEMISTRY, Issue 4 2001SOOTTAWAT BENJAKUL ABSTRACT Cohn's fraction I-S from porcine plasma showed the highest transglutaminase activity, compared to fractions I. 11+III, IV, IV-l. The optimum temperature for incorporating monodancylcadaverine into dimethylated casein was 45C. Plasma transglutaminase in fraction I-S was activated by calcium chloride but was inhibited by N-ethylmaleimide, ethylenediaminetetraacetic acid, and ammonium chloride. The addition of fraction I-S into bigeye snapper surimi resulted in a substantial increase in gel breaking force and deformation, particularly in the presence of calcium chloride and thrombin. No changes in whiteness and water holding capacity were observed in surimi gel with the addition of 0,0.5% of fraction I-S. Fraction I-S was found to catalyze nondisulfide covalent cross-linking of myosin heavy chain. The combination of endogenous and plasma transglutaminase enhanced surimi gelation. [source] Gel Strengthening Effect of Wood Extract on Surimi Produced from Mackerel Stored in IceJOURNAL OF FOOD SCIENCE, Issue 8 2009A.K. Balange ABSTRACT:, The effect of ethanolic kiam wood extract (EKWE) and commercial tannin (CT) on the gel properties of surimi produced from mackerel (Rastrelliger kanagurta) stored in ice for different times (0 to 12 d) was studied. During 12 d of iced storage, pH, total volatile base (TVB), trimethylamine (TMA), and trichloroacetic acid (TCA)-soluble peptide contents as well as thiobarbituric acid-reactive substances (TBARS) of mackerel mince increased while myosin heavy chain (MHC) band intensity decreased continuously (P,< 0.05). The result suggested that deterioration, protein degradation, and lipid oxidation proceeded with increasing storage time. For corresponding surimi, TVB and TMA were almost removed and TBARS and TCA soluble peptide contents were decreased. Conversely, MHC became more concentrated. Decreases in gel-forming ability of surimi were observed when fish used as raw material were stored in ice for a longer time, regardless of EKWE or CT addition. Whiteness of surimi gel decreased and expressible moisture increased especially when the storage time increased. However, superior breaking force and deformation of surimi gel with 0.15% EKWE or 0.30% CT added, compared to those of the control gel were observed during the first 6 d of the storage. Thereafter, EKWE and CT had no gel enhancing effect on surimi. Therefore, freshness was a crucial factor determining gel enhancing ability of EKWE or CT toward mackerel surimi. [source] Application of Recombinant Chum Salmon Cystatin to Alaska Pollock (Theragra chalcogramma) Surimi to Prevent Gel WeakeningJOURNAL OF FOOD SCIENCE, Issue 5 2007D.K. Li ABSTRACT:, Recombinant chum salmon cystatin (RC) expressed in Saccharomyces cerevisiae was purified by His-select nickel affinity chromatography. The specific inhibitory activities of RC against papain and cathepsin L were 7.45 and 10.24 U/mg, respectively. RC was stable over pH 5.0 to 7.0 and at temperature below 65 °C. RC was used to prevent the gel weakening of Alaska pollock surimi. RC at 100 ,g/g showed the highest inhibitory activity against the autolysis of surimi based on the analysis of TCA-soluble peptides. As the concentration of RC increased, both the breaking force and deformation of modori gel greatly increased (P < 0.05). The addition of RC resulted in less expressible drip, which coincided with the increase of whiteness. More myosin heavy chain (MHC) was retained as the addition of RC increased. Therefore, RC could prevent the degradation of proteins in Alaska pollock surimi and was better than egg white (EW). Thus, RC could be applied to Alaska pollock surimi to prevent gel weakening and RC at 100 ,g/g was the optimal concentration. [source] | ||||||||||||||||