scholarly journals WNT/β-Catenin Signaling Regulates Multiple Steps of Myogenesis by Regulating Step-Specific Targets

2015 ◽  
Vol 35 (10) ◽  
pp. 1763-1776 ◽  
Author(s):  
Akiko Suzuki ◽  
Richard C. Pelikan ◽  
Junichi Iwata
Keyword(s):  
Muscle Development ◽  
Myoblast Fusion ◽  
C2c12 Cells ◽  
Biological Processes ◽  
Proliferating Cells ◽  
Spatiotemporal Expression ◽  
Primary Mouse ◽  
Proliferation Activity ◽  
Well Differentiated

Molecules involved in WNT/β-catenin signaling show specific spatiotemporal expression and play vital roles in myogenesis; however, it is still largely unknown how WNT/β-catenin signaling regulates each step of myogenesis. Here, we show that WNT/β-catenin signaling can control diverse biological processes of myogenesis by regulating step-specific molecules. In order to identify the temporally specific roles of WNT/β-catenin signaling molecules in muscle development and homeostasis, we usedin vitroculture systems for both primary mouse myoblasts and C2C12 cells, which can differentiate into myofibers. We found that a blockade of WNT/β-catenin signaling in the proliferating cells decreases proliferation activity, but does not induce cell death, through the regulation of genes cyclin A2 (Ccna2) and cell division cycle 25C (Cdc25c). During muscle differentiation, the inhibition of WNT/β-catenin signaling blocks myoblast fusion through the inhibition of the Fermitin family homolog 2 (Fermt2) gene. Blocking WNT/β-catenin signaling in the well-differentiated myofibers results in the failure of maintenance of their structure by disruption of cadherin/β-catenin/actin complex formation, which plays a crucial role in connecting a myofiber's cytoskeleton to the surrounding extracellular matrix. Thus, our results indicate that WNT/β-catenin signaling can regulate multiple steps of myogenesis, including cell proliferation, myoblast fusion, and homeostasis, by targeting step-specific molecules.

scholarly journals Aint/Tacc3 Is Highly Expressed in Proliferating Mouse Tissues During Development, Spermatogenesis, and Oogenesis

2003 ◽  
Vol 51 (4) ◽  
pp. 455-469 ◽  
Author(s):  
Marjo Aitola ◽  
Christine M. Sadek ◽  
Jan-Åke Gustafsson ◽  
Markku Pelto-Huikko
Keyword(s):  
Adult Mouse ◽  
Coiled Coil ◽  
Proliferating Cells ◽  
Helix Loop Helix ◽  
Aryl Hydrocarbon ◽  
Spatiotemporal Expression ◽  
Mouse Tissues ◽  
Murine Homologue

Aint was originally identified on the basis of its interaction in vitro with the aryl hydrocarbon nuclear receptor translocator (Arnt). Arnt is a common heterodimerization partner in the basic helix-loop–helix (bHLH)-PER-ARNT-SIM (PAS) protein family and is involved in diverse biological functions. These include xenobiotic metabolism, hypoxic response, and circadian rhythm. In addition, Arnt has a crucial role during development. Aint is a member of a growing family of transforming acidic coiled-coil (TACC) proteins and is the murine homologue of human TACC3. Here we report the spatiotemporal expression of Tacc3 mRNA and protein in embryonic, postnatally developing, and adult mouse tissues using in situ hybridization and immunocytochemistry. Tacc3 mRNA was highly expressed in proliferating cells of several organs during murine development. However, the only adult tissues expressing high levels were testis and ovary. Immunocytochemistry revealed that Tacc3 is a nuclear protein. Our results suggest that Tacc3 has an important role in murine development, spermatogenesis, and oogenesis.

scholarly journals Multinucleation of Skeletal Muscle in vitro

1960 ◽  
Vol 7 (3) ◽  
pp. 559-565 ◽  
Author(s):  
Charles R. Capers
Keyword(s):  
Muscle Development ◽  
Horse Serum ◽  
Muscle Fibers ◽  
Time Lapse ◽  
Nuclear Migration ◽  
Myoblast Fusion ◽  
Mitochondrial Activity ◽  
Sequence Of Events ◽  
Membrane Wrinkling

Healthy, mature, spontaneously contracting muscle was cultivated from explants of 13-day chick embryos for periods up to 4 months in the multipurpose chamber (Rose, 1954) using cellophane-strip technique (Rose et al., 1958) with silicone gaskets, Eagle's medium including 10 per cent horse serum reinforced with 300 mg-per cent of glucose, and the teased type of explant. This method provided optically ideal conditions for the study of muscle fibers with oil immersion, phase contrast time-lapse cinematography at 1 frame per minute without apparent damage for periods as long as 10 days. In no case was mitosis, amitosis, or nuclear "budding" observed in the course of muscle development. Multinuclear muscle fibers have been shown with cine technique to result from both myoblast fusion and polar extension of preformed (explanted) muscle tissue. Myoblast fusion was the only demonstrable way of giving rise to multinucleation. Nuclear membrane "wrinkling" was shown to be merely a temporary distortion that occurred during nuclear migration and rotation. It is suggested that this phenomenon may be responsible for numerous reports of amitosis in the genesis of muscle fibers. The histological development of new straps resulted from an orderly sequence of events. Included in these were polar extension, nuclear migration, rotation, and fixation. Following these events there was increased mitochondrial activity, myofibril formation, and cross-banding. Spontaneous contractions were seen throughout the entire course of differentiation in vitro but became more regular and stronger in the later stages.

Cytochemical localization of calcium in myoblasts of the chick embryo myotome

1995 ◽  
Vol 53 ◽  
pp. 1062-1063
Author(s):  
R. González Santander ◽  
M.V. Toledo Lobo ◽  
F.J. Martínez Alonso ◽  
G. Martínez Cuadrado ◽  
M. Gánzalez-Santander Martinez ◽  
...  
Keyword(s):  
Membrane Fusion ◽  
Muscle Development ◽  
Myoblast Fusion ◽  
Cytochemical Localization ◽  
Intracellular Ca ◽  
First Time ◽  
And Storage ◽  
Multinucleated Myotubes

Muscle fibers are derived from multinucleated myotubes which are themselves formed during embryonic development by the fusion of mononucleated myoblasts. Myoblast fusion results from a sequence of different and highly orchestrated stages demonstrated previously in vitro: recognitionalignment, adhesion and membrane fusion. Like many other fusion systems, myoblast fusion is Ca2+ - dependent. The role of Ca2+ is multiple since it is needed for muscle cell differentiation, for the alignment stage and it has also been demonstrated that Ca2+ influx precedes fusion increasing free intracellular Ca2+. It has been proposed that this increase in free intracellular Ca2+ may activate an enzimatic cascade which leads to membrane fusion.The present study, using the K-pyroantimonate method, describes Ca2+ localization and storage in myoblasts before fusion for the first time, since this method had not been applied to skeletal muscle development studies before. Chick embryos from 51 to 108 h. of incubation (Hamburger and Hamilton stages 16 to 25) were used.

scholarly journals Creatine kinase B is necessary to limit myoblast fusion during myogenesis

2015 ◽  
Vol 308 (11) ◽  
pp. C919-C931 ◽  
Author(s):  
Adriana Simionescu-Bankston ◽  
Christophe Pichavant ◽  
James P. Canner ◽  
Luciano H. Apponi ◽  
Yanru Wang ◽  
...  
Keyword(s):  
Creatine Kinase ◽  
Protein Trafficking ◽  
Actin Polymerization ◽  
Muscle Growth ◽  
Cell Types ◽  
Myoblast Fusion ◽  
Biochemical Interaction ◽  
Creatine Kinase B ◽  
Primary Mouse

Myoblast fusion is critical for proper muscle growth and regeneration. During myoblast fusion, the localization of some molecules is spatially restricted; however, the exact reason for such localization is unknown. Creatine kinase B (CKB), which replenishes local ATP pools, localizes near the ends of cultured primary mouse myotubes. To gain insights into the function of CKB, we performed a yeast two-hybrid screen to identify CKB-interacting proteins. We identified molecules with a broad diversity of roles, including actin polymerization, intracellular protein trafficking, and alternative splicing, as well as sarcomeric components. In-depth studies of α-skeletal actin and α-cardiac actin, two predominant muscle actin isoforms, demonstrated their biochemical interaction and partial colocalization with CKB near the ends of myotubes in vitro. In contrast to other cell types, specific knockdown of CKB did not grossly affect actin polymerization in myotubes, suggesting other muscle-specific roles for CKB. Interestingly, knockdown of CKB resulted in significantly increased myoblast fusion and myotube size in vitro, whereas knockdown of creatine kinase M had no effect on these myogenic parameters. Our results suggest that localized CKB plays a key role in myotube formation by limiting myoblast fusion during myogenesis.

scholarly journals In Vitro Effect of CCL11 on Myogenic Differentiation and Its Relevance With Sarcopenia Parameters in Older Adults

2021 ◽  
Vol 5 (Supplement_1) ◽  
pp. A234-A235
Author(s):  
So Jeong Park ◽  
Da Ae Kim ◽  
Jin Young Lee ◽  
Beom-Jun Kim
Keyword(s):  
Older Adults ◽  
Physical Performance ◽  
Myogenic Differentiation ◽  
Muscle Metabolism ◽  
C2c12 Cells ◽  
Differentiation Markers ◽  
Skeletal Muscle Index ◽  
Primary Mouse ◽  
Vitro Effect

Abstract Background: The C-C motif chemokine ligand 11 (CCL11) has been receiving attention as a potential pro-aging factor and thus may be also involved in muscle metabolism and sarcopenia, a key component of aging phenotypes. To clarify this possibility, we investigated the effects of CCL11 on in vitro muscle biology and clinical relevance with sarcopenia parameters in older adults. Methods: Primary mouse myoblasts and C2C12 cells were used for experimental research. Blood samples were collected from 79 participants who underwent a functional assessment, and CCL11 level was measured using a quantikine ELISA kit. Sarcopenia was diagnosed using the Asian-specific cut-off points. Results: CCL11 treatment stimulated the differentiation of both primary myoblasts and C2C12 cells into mature myotubes, and consistently increased the expression of myogenic differentiation markers, such as myosin heavy chain and myogenin. Among C-C chemokine receptors (CCRs), CCR5, not CCR2 and CCR3, was predominantly expressed in muscle cells, and CCR5 inhibitor blocked CCL11-stimulated myogenesis. In a clinical study, after adjustment for sex, age, and body mass index, serum CCL11 level was not significantly different according to the status of sarcopenia, low muscle mass, low muscle strength, and low physical performance, and was not associated with any of skeletal muscle index, grip strength, gait speed, time to complete 5 chair stands, and short physical performance battery score. Conclusions: Contrary to expectations, CCL11 showed the beneficial effects on muscle metabolism at least in vitro system. However, the role of CCL11 on muscle health in humans was not evident, suggesting that circulating CCL11 level may not be a useful biomarker for sarcopenia risk assessment in older adults.

scholarly journals NOTCH inhibition promotes myoblast fusion by releasing HEYL repression on TMEM8C regulatory regions in foetal skeletal muscles

2020 ◽  
Author(s):  
Joana Esteves de Lima ◽  
Cédrine Blavet ◽  
Marie-Ange Bonnin ◽  
Estelle Hirsinger ◽  
Emmanuelle Havis ◽  
...  
Keyword(s):  
Muscle Development ◽  
Close Association ◽  
Myoblast Fusion ◽  
Promoting Effect ◽  
Notch Signalling Pathway ◽  
Regulatory Regions ◽  
Central Regions ◽  
Notch Inhibition

AbstractDifferentiation and fusion are two intricate processes involved in skeletal muscle development. The close association of differentiation and fusion makes it difficult to address the process of fusion independently of differentiation. Using the fusion marker myomaker, named TMEM8C in chicken, we found that both TMEM8C transcripts and the differentiated and fusion-competent MYOG+ cells are preferentially regionalized in the central regions of limb foetal muscles in chicken embryos. Because the NOTCH signalling pathway is a potent inhibitor of muscle differentiation during developmental myogenesis, NOTCH function in myoblast fusion was not addressed so far. We analysed the consequences of NOTCH inhibition for myoblast fusion and TMEM8C expression during foetal myogenesis using in vitro and in vivo chicken systems. NOTCH inhibition following chicken embryo immobilisation or in myoblast cultures increased TMEM8C expression and myoblast fusion. Moreover, we showed that NOTCH inhibition induced the un-binding of the HEYL transcriptional repressor from the TMEM8C regulatory regions in limb muscles and myoblast cultures. These results identify a molecular mechanism underlying the fusion-promoting effect of NOTCH-inhibition during foetal myogenesis.

scholarly journals Trans-regulation of myogenin promoter/enhancer activity by c-ski during skeletal-muscle differentiation: the C-terminus of the c-Ski protein is essential for transcriptional regulatory activity in myotubes

1997 ◽  
Vol 328 (2) ◽  
pp. 607-613 ◽  
Author(s):  
Koji ICHIKAWA ◽  
Takahiro NAGASE ◽  
Shunsuke ISHII ◽  
Akira ASANO ◽  
Naotoshi MIMURA
Keyword(s):  
Terminal Region ◽  
C2c12 Cells ◽  
Upstream Region ◽  
Proliferating Cells ◽  
Mobility Shift ◽  
Enhancer Activity ◽  
C Terminus ◽  
Trans Regulation ◽  
Ski Protein

c-ski gene product is a nuclear protein with myogenesis-promoting and transforming activities. We have analysed the effects of c-ski transfection on the promoter/enhancer activity of the upstream region of the myogenin gene during in vitro myogenesis using CAT reporter assay. When co-transfected with c-ski into myogenic C2C12 cells, promoter/enhancer activity was efficiently suppressed in proliferating cells, but the myogenesis-induced increase in activity was potentiated approximately ten times more (150-fold in the ski-transfected cells) than the ordinary increase (12-fold in the mock) 48 h after induction of differentiation. In non-myogenic 10T1/2 cells, c-ski transfection caused persistent suppression of promoter/enhancer activity in both proliferating and growth-arrested (i.e. myogenesis-inducing) conditions. Thus the ski-dependent potentiation of myogenin gene transcriptional activity appears to be specific for myogenesis. The C-terminal region (amino acids 595-663) of the c-Ski protein was essential for the potentiating activity in myotubes. Other members of the ski-gene family, snoN and snoA, were ineffective in transactivation, possibly because of the defect in the corresponding C-terminal region. c-Ski protein underwent a mobility shift on SDS/PAGE after in vitro myogenesis which may explain the conversion of the activity from suppressive in myoblasts to potentiating in myotubes. Deletion analysis of the upstream region of the myogenin gene revealed that a responsive element to c-ski in myotubes is located at a distinct site upstream of the basal promoter/enhancer region.

scholarly journals Death and Proliferation of Lymphocytes in Immune Response and Tumor Cells Are Controlled by pH Balance

2021 ◽  
Author(s):  
Wei-ping Zeng
Keyword(s):  
Cell Proliferation ◽  
Cell Death ◽  
Unified Theory ◽  
Biological Processes ◽  
Proliferating Cells ◽  
Proliferation Of Lymphocytes ◽  
The Warburg Effect ◽  
Acid Treatments

Many biological processes are controlled by cell death and proliferation. Previous evidence suggests that cell proliferation and death by apoptosis are regulated by separate pathways. The present study found that cellular pH was positively correlated with proliferation but negatively with cell death. Alkaline treatments enhanced lymphocyte proliferation in response to antigen challenge in vivo and in in vitro cultures, whereas acid treatments induced cell death. Low pH was incompatible with the survival of highly proliferating cells, and the susceptibility to the acid-induced death was determined in part by the proliferative status of the lymphocytes. Likewise, alkaline treatments maintained tumor cell proliferation whereas acid treatments induced death. These data support a unified theory for the regulation of cell death and proliferation where a cellular pH balance controls both events, and the mitochondria as proton generators act as pH-stats. Thus, the Warburg effect is viewed as necessary for proliferating cells to have a high cellular pH environment to both survive and accelerate proliferation.

scholarly journals Loss of FilaminC (FLNc) Results in Severe Defects in Myogenesis and Myotube Structure

2006 ◽  
Vol 26 (17) ◽  
pp. 6522-6534 ◽  
Author(s):  
I. Dalkilic ◽  
J. Schienda ◽  
T. G. Thompson ◽  
L. M. Kunkel
Keyword(s):  
Muscle Development ◽  
Structural Integrity ◽  
C2c12 Cells ◽  
Actin Binding ◽  
Unique Role ◽  
Fiber Size ◽  
Fusion Ability ◽  
Dependent Pathway ◽  
Deficient Mice

ABSTRACT FilaminC (FLNc) is the muscle-specific member of a family of actin binding proteins. Although it interacts with many proteins involved in muscular dystrophies, its unique role in muscle is poorly understood. To address this, two models were developed. First, FLNc expression was stably reduced in C2C12 myoblasts by RNA interference. While these cells start differentiation normally, they display defects in differentiation and fusion ability and ultimately form multinucleated “myoballs” rather than maintain elongated morphology. Second, a mouse model carrying a deletion of last 8 exons of Flnc was developed. FLNc-deficient mice die shortly after birth, due to respiratory failure, and have severely reduced birth weights, with fewer muscle fibers and primary myotubes, indicating defects in primary myogenesis. They exhibit variation in fiber size, fibers with centrally located nuclei, and some rounded fibers resembling the in vitro phenotype. The similarity of the phenotype of FLNc-deficient mice to the filamin-interacting TRIO null mice was further confirmed by comparing FLNc-deficient C2C12 cells to TRIO-deficient cells. These data provide the first evidence that FLNc has a crucial role in muscle development and maintenance of muscle structural integrity and suggest the presence of a TRIO-FLNc-dependent pathway in maintaining proper myotube structure.

Regional differences of proliferation activity in the spinal cord ependyma of adult rats

2012 ◽  
Vol 7 (3) ◽  
pp. 397-403 ◽  
Author(s):  
Juraj Blasko ◽  
Marcela Martoncikova ◽  
Kamila Lievajova ◽  
Kamila Saganova ◽  
Andrea Korimova ◽  
...  
Keyword(s):  
Spinal Cord ◽  
Regional Differences ◽  
Central Canal ◽  
Internal Source ◽  
Proliferating Cells ◽  
Adult Rats ◽  
Ki 67 ◽  
Proliferation Activity ◽  
Spinal Cord Regions

AbstractIncreased proliferation activity in the central canal ependyma of adult rodent spinal cord was described after injury and is thought to participate in recovery processes. Proliferation activity is scarce under physiological conditions, but still could be of importance, as in vitro studies showed that the spinal cord ependyma is an internal source of neural stem cells. Data from these studies indicate that there are regional differences in the distribution of proliferation activity along the rostro-caudal axis. We analyzed the proliferation activities in the ependyma within the entire extent of intact adult rat spinal cord. To identify proliferating cells we performed immunohistochemistry either for cell cycle S-phase marker BrdU or for the nuclear protein Ki-67. BrdU and Ki-67 positive cells were counted on sections selected from four spinal cord regions — cervical, thoracic, lumbar and sacral/coccygeal. Analysis showed that the number of BrdU positive cells within the ependyma was very low in all subdivisions of the spinal cord. Both BrdU and Ki-67 labeling revealed a significantly higher number of proliferating cells in the ependyma of sacrococcygeal part in comparison to all other spinal cord regions, suggesting that the caudal spinal cord might have potentially higher regeneration capacity compared to more rostral parts.

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