Titin: structure, isoforms and functional regulation / 生理学报

Titin, the largest known protein in the body expressed in three isoforms (N2A, N2BA and N2B), is essential for muscle structure, force generation, conduction and regulation. Since the 1950s, muscle contraction mechanisms have been explained by the sliding filament theory involving thin and thick muscle filaments, while the contribution of cytoskeleton in force generation and conduction was ignored. With the discovery of insoluble protein residues and large molecular weight proteins in muscle fibers, the third myofilament, titin, has been identified and attracted a lot of interests. The development of single molecule mechanics and gene sequencing technology further contributed to the extensive studies on the arrangement, structure, elastic properties and components of titin in sarcomere. Therefore, this paper reviews the structure, isforms classification, elastic function and regulatory factors of titin, to provide better understanding of titin.

Cholic and deoxycholic acids induce mitochondrial dysfunction, impaired biogenesis and autophagic flux in skeletal muscle cells

BACKGROUND: Skeletal muscle is sensitive to bile acids (BA) because it expresses the TGR5 receptor for BA. Cholic (CA) and deoxycholic (DCA) acids induce a sarcopenia-like phenotype through TGR5-dependent mechanisms. Besides, a mouse model of cholestasis-induced sarcopenia was characterised by increased levels of serum BA and muscle weakness, alterations that are dependent on TGR5 expression. Mitochondrial alterations, such as decreased mitochondrial potential and oxygen consumption rate (OCR), increased mitochondrial reactive oxygen species (mtROS) and unbalanced biogenesis and mitophagy, have not been studied in BA-induced sarcopenia.METHODS: We evaluated the effects of DCA and CA on mitochondrial alterations in C2C12 myotubes and a mouse model of cholestasis-induced sarcopenia. We measured mitochondrial mass by TOM20 levels and mitochondrial DNA; ultrastructural alterations by transmission electronic microscopy; mitochondrial biogenesis by PGC-1α plasmid reporter activity and protein levels by western blot analysis; mitophagy by the co-localisation of the MitoTracker and LysoTracker fluorescent probes; mitochondrial potential by detecting the TMRE probe signal; protein levels of OXPHOS complexes and LC3B by western blot analysis; OCR by Seahorse measures; and mtROS by MitoSOX probe signals. RESULTS: DCA and CA caused a reduction in mitochondrial mass and decreased mitochondrial biogenesis. Interestingly, DCA and CA increased LC3II/LC3I ratio and decreased autophagic flux concordant with raised mitophagosome-like structures. In addition, DCA and CA decreased mitochondrial potential and reduced protein levels in OXPHOS complexes I and II. The results also demonstrated that DCA and CA decreased basal, ATP-linked, FCCP-induced maximal respiration and spare OCR. DCA and CA also reduced the number of cristae. In addition, DCA and CA increased the mtROS. In mice with cholestasis-induced sarcopenia, TOM20, OXPHOS complexes I, II and III, and OCR were diminished. Interestingly, the OCR and OXPHOS complexes were correlated with muscle strength and bile acid levels. CONCLUSION: Our results showed that DCA and CA decreased mitochondrial mass, possibly by reducing mitochondrial biogenesis, which affects mitochondrial function, thereby altering potential OCR and mtROS generation. Some mitochondrial alterations were also observed in a mouse model of cholestasis-induced sarcopenia characterised by increased levels of BA, such as DCA and CA.

iPSCs ameliorate hypoxia-induced autophagy and atrophy in C2C12 myotubes via the AMPK/ULK1 pathway

BACKGROUND: Duchenne muscular dystrophy (DMD) is an X-linked lethal genetic disorder for which there is no effective treatment. Previous studies have shown that stem cell transplantation into mdx mice can promote muscle regeneration and improve muscle function, however, the specific molecular mechanisms remain unclear. DMD suffers varying degrees of hypoxic damage during disease progression. This study aimed to investigate whether induced pluripotent stem cells (iPSCs) have protective effects against hypoxia-induced skeletal muscle injury. RESULTS: In this study, we co-cultured iPSCs with C2C12 myoblasts using a Transwell nested system and placed them in a DG250 anaerobic workstation for oxygen deprivation for 24 h. We found that iPSCs reduced the levels of lactate dehydrogenase and reactive oxygen species and downregulated the mRNA and protein levels of BAX/BCL2 and LC3II/ LC3I in hypoxia-induced C2C12 myoblasts. Meanwhile, iPSCs decreased the mRNA and protein levels of atrogin-1 and MuRF-1 and increased myotube width. Furthermore, iPSCs downregulated the phosphorylation of AMPKA and ULK1 in C2C12 myotubes exposed to hypoxic damage. CONCLUSIONS: Our study showed that iPSCs enhanced the resistance of C2C12 myoblasts to hypoxia and inhibited apoptosis and autophagy in the presence of oxidative stress. Further, iPSCs improved hypoxia-induced autophagy and atrophy of C2C12 myotubes through the AMPK/ULK1 pathway. This study may provide a new theoretical basis for the treatment of muscular dystrophy in stem cells.

Is the myonuclear domain ceiling hypothesis dead?

Muscle fibres are multinuclear cells, and the cytoplasmic territory where a single myonucleus controls transcriptional activity is called the myonuclear domain (MND). MND size shows flexibility during muscle hypertrophy. The MND ceiling hypothesis states that hypertrophy results in the expansion of MND size to an upper limit or MND ceiling, beyond which additional myonuclei via activation of satellite cells are required to support further growth. However, the debate about the MND ceiling hypothesis is far from settled, and various studies show conflicting results about the existence or otherwise of MND ceiling in hypertrophy. The aim of this review is to summarise the literature about the MND ceiling in various settings of hypertrophy and discuss the possible factors contributing to a discrepancy in the literature. We conclude by describing the physiological and clinical significance of the MND ceiling limit in the muscle adaptation process in various physiological and pathological conditions.

Low temperature exposure increases IL-6 expression in skeletal muscle cells / 生理学报

The shivering and nonshivering thermogenesis in skeletal muscles is important for maintaining body temperature in a cold environment. In addition to nervous-humoral regulation, adipose tissue was demonstrated to directly respond to cold in a cell-autonomous manner to produce heat. However, whether skeletal muscle can directly respond to low temperature in an autoregulatory manner is unknown. Transient receptor potential (TRP) channels TRPM8 and TRPA1 are two important cold sensors. In the current study, we found TRPM8 was expressed in mouse skeletal muscle tissue and C2C12 myotubes by RT-PCR. After exposure to 33 °C for 6 h, the gene expression pattern of C2C12 myotubes was significantly changed which was evidenced by RNA-sequencing. KEGG-Pathway enrichment analysis of these differentially expressed genes showed that low temperature changed several important signaling pathways, such as IL-17, TNFα, MAPK, FoxO, Hedgehog, Hippo, Toll-like receptor, Notch, and Wnt signaling pathways. Protein-protein interaction network analysis revealed that IL-6 gene was a key gene which was directly affected by low temperature in skeletal muscle cells. In addition, both mRNA and protein levels of IL-6 were increased by 33 °C exposure in C2C12 myotubes. In conclusion, our findings demonstrated that skeletal muscle cells could directly respond to low temperature, characterized by upregulated expression of IL-6 in skeletal muscle cells.

Muscle architecture of the vastus lateralis and rectus femoris in the production of knee extensor torque in physically inactive women

Abstract Aim: This study aimed to verify the data reliability of muscle architecture (MA) variables, and the relationship between MA and the isometric peak torque (PT) of the monoarticular and biarticular knee extensor (KE) muscle in physically inactive women. Methods: Ten physically inactive women (24.0 ± 1.64 years; 162.9 ± 5.34 cm; 63.5 ± 11.90 kg) participated in the study. An ultrasound device assessed the MA variables (muscle thickness, fascicle length, and pennation angle) of the Vastus Lateralis (VL) and Rectus Femoris (RF), and an isokinetic dynamometer assessed the PT. Pearson correlation evaluated the relationship between PT and MA variables, with a significance level of 5%. Additionally, the intraclass correlation coefficient, coefficient of variation, and standard error of measurement. Results: Excellent reliability between images was observed, and no significant relationships were observed between the PT and MA variables of the VL and RF. Conclusion: Isolated variables of the MA of a monoarticular or a biarticular muscle do not influence the production of the isometric PT of the KE.

Alterações miogênicas em células musculares humanas infectadas pelo vírus zika: susceptibilidade de mioblastos e resistência de miotubos / Myogenic alterations in human muscle cells infected by zika virus: myoblast susceptibility and myotube resistance

O músculo esquelético é responsável pelo movimento e manutenção da postura, e é um órgão produtor de miocinas e altamente metabólico, onde alterações em sua fisiologia podem ter consequências sistêmicas. Esse tecido é alvo para diferentes arboviroses, e mialgia é um sintoma frequentemente relatado. O músculo esquelético é composto majoritariamente por fibras musculares, e uma pequena população de células progenitoras denominadas células satélites (SC), que em caso de lesão podem ser ativadas, proliferam e se diferenciam, sendo capazes de regenerar o tecido muscular. Recentemente nosso grupo demonstrou que SC em proliferação (mioblastos) são infectadas pelo vírus ZIKA (ZIKV), enquanto células diferenciadas e fusionadas (miotubos) não apresentam proteínas virais. O presente trabalho avaliou alterações miogênicas e o perfil transcricional de mioblastos e miotubos humanos após tratamento com ZIKV, com o objetivo de identificar fatores e mecanismos envolvidos na susceptibilidade e resistência destas células à infecção. Confirmamos infecção produtiva do ZIKV nos mioblastos, que apresentaram uma redução no número de células expressando a molécula KI67 em altas concentrações (característico de células em mitose). A análise de sequenciamento mostrou perturbação das vias do ciclo celular em mioblastos infectados, que ainda apresentaram enriquecimento de vias relacionadas à morte celular. Também confirmamos a ausência de infecção produtiva nos miotubos. Interessantemente, verificamos que o ZIKV entra nas células diferenciadas, mas não consegue replicar o RNA viral, e a análise do transcriptoma identificou um enriquecimento de vias e modulação de genes antivirais maior ou exclusivamente nas células diferenciadas em comparação aos mioblastos infectados. Além disso, miotubos expostos ao ZIKV aparentam ter aumento de fusão/hipertrofia. Ao contrário dos mioblastos, miotubos apresentaram enriquecimento de vias relacionadas a organização da matriz extracelular. Dados preliminares do nosso grupo mostraram que o cultivo de mioblastos sobre a isoforma de laminina 511 levou à redução da infecção pelo ZIKV. Contudo, em nossos ensaios, a infecção pelo ZIKV não modulou a expressão de receptores para LM e o bloqueio do receptor de LM, a integrina α6, não reduziu a infecção pelo ZIKV em mioblastos. Nosso trabalho mostrou que a infecção pelo ZIKV induz resposta imune e antiviral, que foram enriquecidos em mioblastos e miotubos, sendo que estes apresentaram uma assinatura única de vias e genes antivirais, que poderiam explicar a resistência frente ao ZIKV.

Study of muscle fibers of the extensor digitorium longus and soleus muscles of C57BL/6 females exposed to glyphosate during pregnancy and lactation / Estudo das fibras musculares dos músculos extensor longo dos dedos e sóleo de fêmeas C57BL/6 expostas ao glifosato durante a prenhez e a lactação

ABSTRACT Objective To evaluate the morphology and morphometry of the muscles extensor digitorium longus and soleus of C57BL/6 females, who were exposed to glyphosate during pregnancy and lactation. Methods Twelve female mice from the C57BL/6 lineage were used. After detection of pregnancy, they were divided into a Control Group, which received only water, and a Glyphosate Group, which received water with 0.5% glyphosate during pregnancy and lactation. Both groups received ad libitum standard diet. After weaning, the females were euthanized and weighed; naso-anal length was measured, and fats were collected and weighed. The muscles extensor digitorium longus and soleus were collected, and their length and weight were measured. Then, the muscles were fixed in Methacarn to perform the histological study of muscle fibers. Results Glyphosate Group presented lower weight gain during pregnancy and also lower final body weight and naso-anal length; however, the other body parameters evaluated did not present a significant difference in relation to the Control Group. Significant differences were also not observed in the analysis of muscle fibers and connective tissue. Conclusion Exposure to 0.5% glyphosate during pregnancy and lactation resulted in lower weight gain during pregnancy, final weight, and naso-anal length. Despite not directly altering the morphology of muscle tissue, these results may indicate enough exposure to interfere with animal metabolism.

RESUMO Objetivo Avaliar a morfologia e a morfometria dos músculos extensor longo dos dedos e sóleo de fêmeas C57BL/6 expostas ao glifosato durante a prenhez e lactação. Métodos Foram utilizados 12 camundongos fêmeas da linhagem C57BL/6. Após detecção da prenhez, foram separadas em Grupo Controle, que recebeu somente água, e Grupo Glifosato, que recebeu água com 0,5% de glifosato durante a prenhez e lactação. Ambos os grupos receberam dieta padrão ad libitum. Após o desmame, as fêmeas foram eutanasiadas e pesadas; o comprimento nasoanal foi mensurado, e as gorduras foram coletadas e pesadas. Os músculos extensor longo dos dedos e sóleo foram coletados, e seu comprimento e peso foram mensurados. Em seguida, os músculos foram fixados em Methacarn para a realização do estudo histológico das fibras musculares. Resultados O Grupo Glifosato apresentou menor ganho de peso durante a prenhez e também menor peso corporal final e comprimento nasoanal, entretanto os demais parâmetros corporais avaliados não apresentaram diferença significativa em relação ao Grupo Controle. Na análise das fibras musculares e do tecido conjuntivo, também não foram observadas diferenças significativas. Conclusão A exposição a 0,5% de glifosato durante a prenhez e lactação resultou em menor ganho de peso na gestação, peso final e comprimento nasoanal, o que pode indicar que, apesar de não alterar a morfologia do tecido muscular diretamente, a exposição foi suficiente para interferir no metabolismo dos animais.

Tipo de fibra muscular y su relación con el abordaje fonoaudiológico en los trastornos de la deglución / Type of muscle fibreand its relationship to the Speech-Language Pathology approach in swallowing disorders

Los actuales protocolos de intervención en deglución no especifican parámetros de los ejercicios tales como la cantidad de series, duración, tiempos de repetición, entre otros. Lo anterior, es un desafío para la intervención fonoaudiológica en los trastornos de la deglución. El objetivo de este trabajo es acercar al profesional fonoaudiólogo al conocimiento de las fibras musculares, ya que es un conocimiento que se debe considerar antes de indicar ejercicios musculares cráneo-cérvico-orales. Por ello, se describen las principales fibras musculares con sus respectivas características fundamentales, como lo son: la resistencia a la fatiga y la velocidad de contracción. Se revisa la literatura sobre la dispersión de las fibras musculares, de algunos de los principales músculos que participan en el proceso deglutorio. También, se analizan diversas particularidades de los músculos de la zona cráneo-cérvico-oral. Se describen además las diferentes dificultades para evaluar esta musculatura. Finalmente, se expone la relevancia práctica de conocer estos tipos de fibras musculares y las perspectivas futuras de este enfoque basado en parámetros del ejercicio y la medición de variables objetivas.

Swallowing intervention protocols in dysphagia do not specify the parameters of the exercises, namely the duration, frequencyand number of series, all of which present a challenge in speech therapy intervention in swallowing disorders. The purpose of this review is to provide Speech and Language Pathologists with background information concerning crucial muscle fibres used in swallowing therapy and to subsequently indicate appropriate skull-cervical-oral muscle exercises. We describe the primary muscle and its characteristics such as fatigue resistance and contraction speed, review the types of fibres of some of the main muscles that participate in the swallowing process, and present the particularities of the muscles of the cranio-cervical-oral area in comparison with the rest of the skeletal muscles. We also provide information regarding the difficulties in evaluating the musculature involvedin the swallowing process before finally highlighting the relevance of understanding the roles and characteristics of these muscles for clinical practice

Plasticidad muscular y fibras híbridas en la musculatura masticatoria: revisión de literatura / Muscle plasticity and hybrid fibers in the masticatory musculature: literature review

La musculatura masticatoria se caracteriza por presentar fibras híbridas que en los últimos años se han relacionado con el fenómeno de plasticidad muscular. El objetivo del estudio fue describir la relación entre la plasticidad muscular y las fibras musculares híbridas presentes en la musculatura masticatoria, mediante una revisión narrativa de literatura. Para esto, se realizó una búsqueda electrónica en PUBMED, ScienceDirect y BIREME, utilizando las palabras claves: "Muscle Plasticity", "Hybrid Muscle Fibers" y "Hybrid Fibers". Fueron seleccionados documentos que reportan las isoformas de cadena pesada de miosina (MHC) presentes en los músculos masticatorios de humanos y otros mamíferos, junto a los cambios vinculados a demandas funcionales. Se describe la presencia de fibras puras tipo I y tipo II, además de otras isoformas como la MHC-la, MHC-IIM, MHC-fetal y MHC-cardíaca. Sin embargo, un porcentaje considerable de fibras en la musculatura masticatoria son híbridas, es decir, expresan a más de una isoforma de MHC, las cuales también son diferentes a nivel intermuscular e intramuscular. Las influencias locales pueden contribuir a la variación de la expresión del tipo de fibra. En la musculatura masticatoria, el destete, la dureza de los alimentos, el bruxismo, la morfología craneofacial y el uso de prótesis dentales genera cambios a nivel de los músculos masticatorios, donde es común la presencia de fibras híbridas. Se concluye la presencia importante de fibras híbridas en la musculatura masticatoria y su relación con la plasticidad muscular a lo largo del ciclo vital, debido a cambios funcionales y patológicos. Es importante que los terapeutas de habla y motricidad orofacial profundicen en el conocimiento de la fisiología del comportamiento oromiofuncional.

The masticatory musculature is characterized by presenting hybrid fibers that in recent years have been related to the phenomenon of muscle plasticity. The objective of the study was to describe the relationship between muscle plasticity and the hybrid muscle fibers present in the masticatory muscles, through a narrative review. For this, an electronic search was conducted in PUBMED, ScienceDirect and BIREME, using the keywords: "Muscle Plasticity", "Hybrid Muscle Fibers" and "Hybrid Fibers". Documents that report the myosin heavy chain (MHC) isoforms present in the masticatory muscles of humans and other mammals were selected, along with the changes linked to functional demands. The presence of type I and type II pure fibers were described, in addition to other isoforms such as MHC-la, MHC-IIM, MHC-fetal and MHC-cardiac. However, a significant percentage of fibers in the masticatory muscles are hybrids, that is, they express more than one MHC isoform, which are also different at the intermuscular and intramuscular level. Local influences can contribute to the variation of fiber type expression. In the chewing muscles, weaning, the hardness of food, bruxism, craniofacial morphology and the use of dental prostheses generate changes at the level of the chewing muscles, where the presence of hybrid fibers is common. The important presence of hybrid fibers in the masticatory muscles and their relationship with muscle plasticity throughout the life cycle, due to functional and pathological changes, is concluded. It is important for Speech Therapy andMyofunctional Therapy to deepen their understanding of the physiology of oromyofunctional behavior.

La rigidez cadavérica, el espasmo cadavérico y tipos de fibras musculares / The cadaveric rigidity, the cadaveric spasm and types of muscle fibers

La rigidez cadavérica (rigor mortis) es un proceso no muy bien comprendido por la mayoría de los médicos. El conocimiento de la intimidad del proceso de la rigidez cadavérica es de vital importancia ya que es una de las variables que junto con las livideces (livor mortis) y la temperatura (algor mortis) del cadáver ayudan a determinar el cronotanatodiagnóstico, tanatocronodiagnóstico o intervalo postmortal del período inmediato de la muerte. Para entender el mecanismo de la rigidez y el espasmo cadavérico es preciso hacer un repaso de la contracción muscular fisiológica en el vivo. Hay que tener presente que el tipo de fibra muscular predominante modificará las características de la contracción muscular fisiológica en el vivo, y también la rigidez y el espasmo cadavérico. (AU)

The cadaveric rigidity (rigor mortis) is a process which is not very well understood by the majority of the doctors. The knowledge of the intimacy of the cadaveric stiffness process is of vital importance since it is one of the variables that, as well as the postmortem lividity (livor mortis) and the body temperature post mortem (algor mortis) help determine the chronotanatodiagnostic, tanatochronodiagnostic or postmortal interval of the immediate period of death. In order to understand the mechanism of stiffness and cadaveric spasm, it is necessary to review the physiological muscle contraction in vivo. We should keep in mind that the predominant type of muscle fiber will modify the characteristics of physiological muscle contraction in vivo, as well as stiffness and cadaveric spasm. (AU)

Análise da ativação muscular durante o movimento de alcance nas condições ativo, ativo-assistido e autoassistido em pacientes pós-AVE / Análisis de la activación muscular durante el movimiento de alcance en las condiciones activo, activo-asistido y auto-asistido en pacientes post-AVE / Analysis of muscle activation during reaching movement in active, active-assisted and self-assisted conditions in post-stroke patients

RESUMO O Acidente Vascular Encefálico (AVE) é uma patologia que frequentemente causa limitações motoras nos Membros Superiores (MMSS) gerando prejuízos funcionais nos movimentos de alcance. O objetivo do estudo foi analisar o recrutamento muscular do membro superior parético durante três condições de alcance: ativo, ativo-assistido e autoassistido, através de dados eletromiográficos das fibras anteriores do Músculo Deltoide (MD), Bíceps Braquial (BB) e Tríceps Braquial (TB). Estudo do tipo transversal que utilizou como testes clínicos o miniexame do estado mental, escala de equilíbrio de Berg, medida de independência funcional, escala modificada de Ashworth e escala de Fugl-Meyer - seção MMSS. A coleta dos dados eletromiográficos de superfície foi realizada utilizando-se o eletromiógrafo e eletrodos de configuração bipolar da EMG System do Brasil com três canais posicionados nos pontos motores do MD (fibras anteriores), BB e TB de ambos os membros superiores. As variáveis clínicas apresentaram resultados de comprometimento motor, cognitivo e funcional leves. Os dados eletromiográficos mostraram que o MD e TB durante o alcance ativo-assistido contraíram mais que no alcance autoassistido (p<0.05). Os MD e TB apresentaram diferenças significativas durante os movimentos de alcance, enquanto que o músculo BB não mostrou alterações. Entre os diversos tipos de alcance, o ativo-assistido foi o que proporcionou maior ativação muscular. Sugere-se que sejam feitos ensaios clínicos para verificar a eficácia dos treinamentos.

RESUMEN El Accidente Vascular Encefálico (AVE) es una patología que frecuentemente causa limitaciones motoras en los Miembros Superiores (MMSS) generando perjuicios funcionales en los movimientos de alcance. El objetivo del estudio fue analizar el reclutamiento muscular del miembro superior parético durante tres condiciones de alcance: activo, activo-asistido y auto-asistido, a través de datos electromiográficos de las fibras anteriores del Músculo Deltóide (MD), Bíceps Braquial (BB) y Tríceps Braquial (TB). Estudio del tipo transversal que utilizó como pruebas clínicas el mini-examen del estado mental, escala de equilibrio de Berg, medida de independencia funcional, escala modificada de Ashworth y escala de Fugl-Meyer - sección MMSS. La recolección de los datos electromiográficos de superficie fue realizada utilizando el electromiografo y electrodos de configuración bipolar de la EMG System de Brasil con tres canales colocados en los puntos motores del MD (fibras anteriores), BB y TB de ambos miembros superiores. Las variables clínicas presentaron resultados de compromiso motor, cognitivo y funcional leves. Los datos electromiográficos mostraron que el MD y el TB durante el alcance activo-asistido contrajeron más que en el alcance auto-asistido (p<0.05). Los MD y TB presentaron diferencias significativas durante los movimientos de alcance, mientras que el músculo BB no mostró alteraciones. Entre los diversos tipos de alcance, el activo asistido fue el que proporcionó mayor activación muscular. Se sugiere que se realicen ensayos clínicos para verificar la eficacia de los entrenamientos.

ABSTRACT A cerebrovascular accident (CVA) is a disease that often causes upper limb motor limitations and functional losses in reaching movements. The objective of this study was to analyze the muscle recruitment of the paretic upper limb during three reaching conditions: active, active-assisted and self-assisted, through electromyographic data of anterior fibers of Deltoid Muscle (DM), Biceps Brachii (BB) and Triceps Brachii (TB). Cross-sectional study that used as clinical trials the Mini-Mental State Examination, Berg balance scale, functional independence measure, the modified Ashworth scale, and the Fugl-Meyer assessment - upper limbs section. Surface electromyographic data were collected using the electromyograph and bipolar electrode configuration of the EMG System do Brasil with three channels positioned in the motor points of DM (anterior fibers), BB and TB of both upper limbs. Clinical variables showed mild motor, cognitive, and functional impairment. Electromyographic data showed that DM and TB contracted more during active-assisted than during self-assisted exercise (p<0.05). DM and TB presented significant differences during reaching movements, while the BB muscle showed no changes. Among the different reaching exercises, the active-assisted was the one that provided greater muscle activation. Clinical trials are suggested to verify the effectiveness of the training.

Correlation analysis between the surface electromyography and muscle fiber types of the core muscle group in the patients with myofascial pain syndromes / 中国骨伤

OBJECTIVE@#To detect the core muscle group in the patients with myofascial pain syndromes(MPS) by using the surface electromyography; to detect the distribution of muscle fiber type by the analysis of the median frequency and the slope of the median frequency.@*METHODS@#From October 2017 to March 2018, there were 100 patients with the MPS, including 45 males and 55 females; the average age was 48.5 years old, ranging from 29 to 76 years old. There were 40 cases of left back pain and 60 cases of right back pain. The course of illness was more than 6 months. Another 40 healthy patients without pain in the waist were included in the control group, 20 males and 20 females; the average age was 47.3 years old, ranging from 29 to 76 years old. All the patients had different degrees of back pain and muscle stiffness, which were diagnosed as lumbar fasciitis by clinical and imaging examination. Surface electromyography was used to measure the characteristics of the lumbar core muscles (multifissions, iliocostal muscles, and longest muscle) of the three groups in the Biering-Sorensen testing, such as median frequency(MF) and absolute slope of median frequency (MFs).@*RESULTS@#The MF values of the multifidus muscle in the three groups were as follows:the left side of the non-pain group was 133.88±26.61, and the right side was 131.39±29.81; left side of lift side pain group 117.29±10.93, right side 133.70±17.81; in the right pain group, the left side was 131.36±17.37, and the right side was 118.28±13.57. The MF values of the iliocostal muscle in the three groups were:106.94±28.01 on the left side of the non-pain group, 114.68±18.96 on the right side; left side of lift side pain group 93.95±11.17, right side 107.60±27.86; in the right pain group, the left side was 105.93±15.52, and the right side was 97.27±19.27. The MF values of the longest muscle in the three groups were:109.24±26.20 on the left side of the non-pain group, 112.58±17.70 on the right side. Left side of left side pain group 95.58±10.83, right side 108.79±26.39; in the right pain group, the left side was 106.50±17.98, and the right side was 98.20±11.16. The MFs values of the multifidus muscle in the three groups were:0.221±0.109 on the left side of the non-pain group, and 0.259±0.169 on the right side; left side of left side pain group 0.318±0.184, right side 0.210±0.159; in the right pain group, the left side was 0.258±0.169, and the right side was 0.386±0.166. The MFs values of the iliocostal muscles in the three groups were:0.241±0.158 for the left side of the non-pain group, and 0.238±0.128 for the right side. Left side of left side pain group 0.330±0.208, right side 0.252±0.171; in the right side pain group, left side 0.249±0.150, right side 0.343± 0.144. The MFs values of the longest muscle of the three groups were:0.244±0.252 on the left side of the non-pain group, and 0.210±0.128 on the right side; left side of left side pain group 0.348±0.255, right side 0.241±0.224; in the right pain group, the left side was 0.239±0.155, and the right side was 0.334±0.233. There were no statistically significant differences in MF and MFs values of the left and right lumbar multifidus muscle, iliocostal muscle and longest muscle in the non-pain group(>0.05). MF values of the pain side multifidus muscle, iliocostal muscle and longest muscle in the lumbago group were lower than those in the non-pain group(<0.05). MFs values of the painful side multifidus muscle, iliocostal muscle and longest muscle in the low back pain group were higher than those in the non-pain group(<0.05).@*CONCLUSIONS@#The muscle fatigue degree of the back muscle in the pain side of patients with MPs is decreased, and the muscle fiber type is dominated by II muscle fiber.

Expression of EGR1 gene and location of EGR1 protein in differentiation of bovine skeletal muscle-derived satellite cells / 中国应用生理学杂志

OBJECTIVE@#To investigate the expression of EGR1 gene and the localization of EGR1 protein in bovine skeletal muscle-derived satellite cells (MDSCs), as well as to investigate the mechanism that EGR1 protein enters the nucleus.@*METHODS@#Bovine MDSCs were cultured in differentiation medium for 1 day, 3 days and 5 days, respectively, and each group was triplicate. The expression of EGR1 gene and the localization of EGR1 protein were studied at different differentiation period in MDSCs by qRT-PC and Western blot. Moreover, the changes on the expression of endogenous EGR1 gene and EGR1 proteins were explored by CRISPRi, site-directed mutagenesis and laser confocal method.@*RESULTS@#The results from the qRT-PCR and Western blot showed that the expressions of EGR1 gene on transcription level and translation level were significantly higher in differentiated cells than those in undifferentiated cells. The highest expression was found on the third day after the differentiation, and then began to decline. Immunofluorescence assays showed that EGR1 proteins were preferentially expressed in differentiated MDSCs, and increased along with the increase of number of myotubes. Confocal observation revealed that some EGR1 proteins were transferred into the nucleus in the differentiation of cells, however, the EGR1 proteins would not be detected in the differentiated MDSCs nuclei if a site directed mutagenesis (serine) on EGR1 protein occurred.@*CONCLUSION@#During the differentiation of bovine skeletal muscle satellite cells, the transcriptional level of EGR1 gene is increased, and some EGR1 proteins are transferred into the nucleus. The serine phosphorylation at position 533 of the C terminal of EGR1 protein is necessary for the nucleus transfer.

Therapeutic effect of massage on denervated skeletal muscle atrophy in rats and its mechanism / 中国应用生理学杂志

OBJECTIVE@#To investigate the therapeutic effects of massage on denervated skeletal muscle atrophy in rats and its mechanism.@*METHODS@#Forty-eight male SD rats were randomly divided into model group (n=24) and massage group (n=24). Gastrocnemius muscle atrophy model was established by transecting the right tibial nerve of rat. On the second day after operation, the gastrocnemius muscle of the rats in the massage group was given manual intervention and the model group was not intervened. Six rats were sacrificed at the four time points of 0 d, 7 d, 14 d and 21 d. The gastrocnemius of the rats were obtained and measured the wet mass ratio after weighing. Cross-sectional area and diameter of the muscle fiber were measured after HE staining. The relative expressions of miR-23a, Akt, MuRF1 and MAFbx mRNA were tested with qPCR.@*RESULTS@#Compared with 0 d, the wet weight ratio, cross-sectional area and diameter of gastrocnemius muscle showed a progressive decline in the model group and massage group. The wet weight ratio, cross-sectional area and diameter of gastrocnemius muscle in the massage group were higher than those in the model group on 7 d, 14 d and 21 d (P＜0.05, P＜0.01). Compared with 0 d, the expressions of MuRF1, MAFbx and Akt mRNA were increased first and then were decreased in the model group and massage group. The expression of MuRF1 mRNA in massage group was lower than that in model group on 7 d and 21 d (P＜0.05, P＜0.01). The expression of MAFbx mRNA in massage group was lower than that in model group on 7 d, 14 d and 21 d (P＜0.01, P＜0.05, P＜0.01). The expression of Akt mRNA in massage group was higher than that in model group on 7 d, 14 d and 21 d (P＜0.05, P＜0.01). Compared with 0 d, the expression of miR-23a mRNA was increased in the model group and massage group on 21 d, and the expression of miR-23a mRNA in massage group was higher than that in model group (P＜ 0.05).@*CONCLUSION@#Massage can delay the atrophy of denervated skeletal muscle. The mechanism may be related to up-regulation of the expression of miR-23a and Akt mRNA, down-regulation of the expressions of MuRF1 and MAFbx mRNA, inhibition of protein degradation rate, and reduction of skeletal muscle protein degradation.

Muscle changes with high-intensity aerobic training in an animal model of renal disease

Abstract Purpose: To analyze the muscle changes with high-intensity aerobic training (HIAT) in an animal model of renal disease (RD). Methods: Twenty one adult Wistar rats were divided into 3 groups: healthy sedentary (HS), RD sedentary (RDS), RD aerobic training (RDAT). RDS and RDAT were subjected to unilateral renal ischemia-reperfusion (10 min) and 21days after that, RDAT was subjected to 6 weeks HIAT (swimming). Serum creatinine (Cr) and muscle morphometry (cross-sectional area = CSA) of gastrocnemius were analyzed. Results: Cr was higher (p = 0.0053) in RDS (0.82 ± 0.04) than in the others (RDAT 0.55 ± 0.04; HS 0.55 ± 0.04). Morphometric analysis (class interval of CSA in μm2/absolute frequency of muscle fibers in each class) indicated that 50th percentile occurred in: HS 7th class (3000.00-3499.00/515), RDS, 8th class (3500.00-3999.00/484), RDAT 5th class (2000.00-2499.00/856). CSA of largest fibers in RDS, RDAT, HS was 9953.00 μm2, 9969.00 μm2,11228.00 μm2, respectively. High frequency of fibers with lower CSA occurred in 4th, 5th, 6th and 7th class in RDA, absence of fibers into 22nd, 23rd classes (RDS and RDAT). Conclusion: HIAT in an animal model of RD resulted in increased the number of muscle fibers with smaller CSA.

Deficiency of Anoctamin 5/TMEM16E causes nuclear positioning defect and impairs Ca²⁺ signaling of differentiated C2C12 myotubes

Anoctamin 5 (ANO5)/TMEM16E belongs to a member of the ANO/TMEM16 family member of anion channels. However, it is a matter of debate whether ANO5 functions as a genuine plasma membrane chloride channel. It has been recognized that mutations in the ANO5 gene cause many skeletal muscle diseases such as limb girdle muscular dystrophy type 2L (LGMD2L) and Miyoshi muscular dystrophy type 3 (MMD3) in human. However, the molecular mechanisms of the skeletal myopathies caused by ANO5 defects are poorly understood. To understand the role of ANO5 in skeletal muscle development and function, we silenced the ANO5 gene in C2C12 myoblasts and evaluated whether it impairs myogenesis and myotube function. ANO5 knockdown (ANO5-KD) by shRNA resulted in clustered or aggregated nuclei at the body of myotubes without affecting differentiation or myotube formation. Nuclear positioning defect of ANO5-KD myotubes was accompanied with reduced expression of Kif5b protein, a kinesin-related motor protein that controls nuclear transport during myogenesis. ANO5-KD impaired depolarization-induced [Ca²⁺]i transient and reduced sarcoplasmic reticulum (SR) Ca²⁺ storage. ANO5-KD resulted in reduced protein expression of the dihydropyridine receptor (DHPR) and SR Ca²⁺-ATPase subtype 1. In addition, ANO5-KD compromised co-localization between DHPR and ryanodine receptor subtype 1. It is concluded that ANO5-KD causes nuclear positioning defect by reduction of Kif5b expression, and compromises Ca²⁺ signaling by downregulating the expression of DHPR and SERCA proteins.

Morphological analysis of articularis genus and involvement in muscle synchronicity with vastus intermedius / Análisis morfológico del músculo articular de la rodilla y su participación en la sincronicidad muscular con el músculo vasto intermedio

The articularis genus, which lies under the vastus intermedius, has been regarded as part of the quadriceps femoris. However, they are well known to have different function because their respective origins and insertions are mutually distinct. These muscles are considered to work almost simultaneously when the knee is extended. The electromyogram has been used to demonstrate muscle co-contraction. However, the articularis genus is deeper than other muscles. Moreover, it is difficult to analyze it by surface electromyogram. The relative proportions of muscle fiber types and the characteristics of these fiber types are important determinants of the surface electromyogram. Furthermore, biomechanical analysis of AG has remained unclear. This study investigated the ratio of muscle fiber types in these muscles. Muscle samples from seven human cadaveric specimens were used with application of immunofluorescence double staining. Results show that in the vastus intermedius and articularis genus, the percentage of Type I fibers was significantly higher than that of Type II fibers. No significant difference was found in the mean percentages of Type I and Type II fiber types. The percentages of Type I and Type II fibers in articularis genus muscle were correlated positively to the percentage in the vastus intermedius. These results suggest that similar muscle fiber compositions of these muscles might reflect their contraction during the same active phase of knee extension, despite their different functions.

El músculo articular de la rodilla, que se encuentra cubierto por el músculo vasto intermedio, se ha considerado como parte del músculo cuádriceps femoral. Sin embargo, es sabido que tienen diferentes funciones debido a que sus respectivos orígenes e inserciones son mutuamente distintas. Se considera que estos músculos trabajan de forma casi simultánea cuando la rodilla está extendida. El electromiograma se ha usado para demostrar la contracción muscular. Sin embargo, el músculo articular de la rodillas es más profundo que otros músculos. Además, es difícil analizarlo por electromiograma de superficie. Las proporciones relativas de los tipos de fibras musculares y las características de estos tipos de fibras son importantes determinantes del electromiograma de superficie. Además, el análisis biomecánico de músculo articular de la rodilla no ha sido claro. Este estudio investigó la proporción de tipos de fibras musculares en estos músculos. Se usaron muestras musculares de siete cadáveres humanos con la aplicación de doble tinción de inmunofluorescencia. Los resultados muestran que en los músculos articular de la rodilla y vasto intermedio, el porcentaje de fibras de Tipo I fue significativamente mayor que el de las fibras de Tipo II. No se encontraron diferencias significativas en los porcentajes medios de los Tipo I y Tipo II. Los porcentajes de fibras Tipo I y Tipo II en el músculo articular de la rodilla se correlacionaron positivamente con el porcentaje en el músculo vasto intermediario. Estos resultados sugieren que las composiciones de las fibras musculares similares de estos músculos podrían reflejar su contracción durante la misma fase activa de la extensión de la rodilla, a pesar de sus diferentes funciones.

Morphological Characterization of Diaphragm in Common Squirrel Monkey (Saimiri sciureus)

ABSTRACT The wall of the diaphragm can be affected by congenital or acquired alterations which allow the passage of viscera between the abdominal and chest cavities, allowing the formation of a diaphragmatic hernia. We characterized morphology and performed biometrics of the diaphragm in the common squirrel monkey Saimiri sciureus. After fixation, muscle fragments were collected and processed for optical microscopy. In this species the diaphragm muscle is attached to the lung by phrenopericardial ligament. It is also connected to the liver via the coronary and falciform ligaments. The muscle is composed of three segments in total: 1) sternal; 2) costal, and 3) a segment consisting of right and left diaphragmatic pillars. The anatomical structures analyzed were similar to those reported for other mammals. Histological analysis revealed stable, organized muscle fibers with alternation of light and dark streaks, indicating transverse striation.

Morphological changes in rat rectus abdominis muscle induced by diabetes and pregnancy

The urethral muscle of diabetic pregnant rats is affected by long-term mild diabetes and short-term severe diabetes, which plays a crucial role in the pathogenesis of pelvic floor disorders. We hypothesized that muscles outside the pelvis are subject to similar changes. The current study aimed at analyzing the effects of long-term mild and short-term severe diabetes on the structure and ultrastructure of fiber muscles and collagen in rats' rectus abdominis (RA) muscle. Therefore, the RA muscle of virgin, pregnant, long-term mild diabetic, short-term severe diabetic, long-term mild diabetic pregnant and short-term severe diabetic pregnant 3-month-old Wistar rats were collected. The structure was analyzed by picrosirius red staining, immunohistochemistry for fast and slow muscle fibers and transmission electron microscopy. We investigated two levels of STZ- induced diabetes: long-term mild diabetes (blood glucose level: 120-200 mg/dL) and short-term severe diabetes (blood glucose level >300 mg/dL). Long-term mild diabetic pregnant and short-term severe diabetic pregnant rats had decreased fast fibers and increased slow fibers, disrupted areas of sarcomere, intermyofibrillar mitochondria and myelin figures in the RA muscle. Both groups enabled us to analyze the specific influence of pregnancy, separately from diabetes. The current study demonstrated that diabetes and pregnancy induced intramuscular transformation and reorganization of RA muscle with a switch of fiber type adjusting their architecture according to intensity and duration of hyperglycemic insult within pregnancy.