• Title/Summary/Keyword: C2C12 myoblast

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Effect of trichostatin A on NF-κB DNA binding activity and myogenesis in C2Cl2 skeletal muscle Precursor cell (C2C12 근육아세포에서 trichostatin A에 의한 NF-κB DNA 결합 활성과 근육발생에 미치는 영향)

  • 임운기;김경창;신혜자
    • Journal of Life Science
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    • v.12 no.1
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    • pp.55-60
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    • 2002
  • The differentiation of skeletal muscle precursor cells in culture is marked by the transcriptional activation of muscle-specific genes and the morphological differentiation of myoblast into multinucleate myotube. In this study, we examined the effect of TSA (Trichostatin A) on WF-kB DNA binding activity and muscle cell fusion in the process of myogenesis. Under TSA treatment, C2C12 myoblast could not fuse to myotube and its NF-kB DNA binding activity was also blocked. To investigate whether these phenomenons were affected by TSA in direct or not, differentiation media (DM) used to differentiate cells without TSA was concentrated and added to C2C12 myoblast with TSA simultaneously. C2C12 myoblast was fused to myotube and NF-kB DNA binding activity was recovered. These results suggest that TSA affects on the differentiation of myoblast, perhaps through several factors, by inhibiting myoblst fusion and blocking NF-kB DNA binding activity.

Setdb1 Is Required for Myogenic Differentiation of C2C12 Myoblast Cells via Maintenance of MyoD Expression

  • Song, Young Joon;Choi, Jang Hyun;Lee, Hansol
    • Molecules and Cells
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    • v.38 no.4
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    • pp.362-372
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    • 2015
  • Setdb1, an H3-K9 specific histone methyltransferase, is associated with transcriptional silencing of euchromatic genes through chromatin modification. Functions of Setdb1 during development have been extensively studied in embryonic and mesenchymal stem cells as well as neurogenic progenitor cells. But the role of Sedtdb1 in myogenic differentiation remains unknown. In this study, we report that Setdb1 is required for myogenic potential of C2C12 myoblast cells through maintaining the expressions of MyoD and muscle-specific genes. We find that reduced Setdb1 expression in C2C12 myoblast cells severely delayed differentiation of C2C12 myoblast cells, whereas exogenous Setdb1 expression had little effect on. Gene expression profiling analysis using oligonucleotide microarray and RNA-Seq technologies demonstrated that depletion of Setdb1 results in downregulation of MyoD as well as the components of muscle fiber in proliferating C2C12 cells. In addition, exogenous expression of MyoD reversed transcriptional repression of MyoD promoter-driven luciferase reporter by Setdb1 shRNA and rescued myogenic differentiation of C2C12 myoblast cells depleted of endogenous Setdb1. Taken together, these results provide new insights into how levels of key myogenic regulators are maintained prior to induction of differentiation.

A Possible Role of Kainate Receptors in C2C12 Skeletal Myogenic Cells

  • Park, Jae-Yong;Han, Jae-Hee;Hong, Seong-Geun
    • The Korean Journal of Physiology and Pharmacology
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    • v.7 no.6
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    • pp.375-379
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    • 2003
  • $Ca^{2+}$ influx appears to be important for triggering myoblast fusion. It remains, however, unclear how $Ca^{2+}$ influx rises prior to myoblast fusion. Recently, several studies suggested that NMDA receptors may be involved in $Ca^{2+}$ mobilization of muscle, and that $Ca^{2+}$ influx is mediated by NMDA receptors in C2C12 myoblasts. Here, we report that other types of ionotropic glutamate receptors, non-NMDA receptors (AMPA and KA receptors), are also involved in $Ca^{2+}$ influx in myoblasts. To explore which subtypes of non-NMDA receptors are expressed in C2C12 myogenic cells, RT-PCR was performed, and the results revealed that KA receptor subunits were expressed in both myoblasts and myotubes. However, AMPA receptor was not detected in myoblasts but expressed in myotubes. Using a $Ca^{2+}$ imaging system, $Ca^{2+}$ influx mediated by these receptors was directly measured in a single myoblast cell. Intracellular $Ca^{2+}$ level was increased by KA, but not by AMPA. These results were consistent with RT-PCR data. In addition, KA-induced intracellular $Ca^{2+}$ increase was completely suppressed by treatment of nifedifine, a L-type $Ca^{2+}$ channel blocker. Furthermore, KA stimulated myoblast fusion in a dose-dependent manner. CNQX inhibited not only KA-induced myoblast fusion but also spontaneous myoblast fusion. Therefore, these results suggest that KA receptors are involved in intracellular $Ca^{2+}$ increase in myoblasts and then may play an important role in myoblast fusion.

Effect of Simple Formulas of Muscle Section in Donguibogam on Myogenic Regulatory Factors and IGF-1 Expression in C2C12 Cells

  • Yang, In Jun;Tettey, Clement;Shin, Heung Mook
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.28 no.4
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    • pp.411-416
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    • 2014
  • Simple formulas (單方) of muscle section in Donguibogam (東醫寶鑑) have long been prescribed for strengthening muscle and/or prevention of age-related muscle loss. However, biological activity and mechanisms by which they influence myoblast differentiation have not been studied. Therefore, in this study, we evaluated the effects of 14 simple formulas on myoblast differentiation in C2C12 myoblast cells under non-cytotoxic ($0.5mg/m{\ell}$) conditions. C2C12 cells were treated with water extracts of simple formulas for 72 h, and RT-PCR was performed to determine the gene expression levels of myogenic regulatory factors (MRFs), including myoD, myogenin, MRF4, myf5, and insulin like growth factor-1 (IGF-1). Treatment with Colocasiae Rhizoma (CR), Pini Semen (PS), and Sesami Semen (SS) resulted in a significant increase in expression of myogenin in C2C12 cells. Treatment with Allii Macrostemi Bulbus (AM), Colocasiae Rhizoma (CR), and Pini Semen (PS) also resulted in increased expression of MRF4 in C2C12 cells. In addition, enhanced expression of IGF-1 was observed in treatment with Eucommiae cortex (EC), Dioscoreae Rhizoma (DR), Colocasiae Rhizoma (CR), Pini Semen (PS), and Sesami Semen (SS) in C2C12 cells. These results indicate that simple formulas of muscle section in Donguibogam could potentially enhance myoblast differentiation at least in part via increasing expression of myogenin, and/or MRF4 and/or IGF-1.

Effects of Histone Deacetylase Inhibitor, Trichostatin A, on the Differentiation of C2C12 Myoblasts and the Expression of Cell Cycle Regulators (히스톤 탈아세틸화 효소 억제제 trichostatin A가 C2C12 myoblast 세포 분화와 세포주기 조절인자의 발현에 미치는 영향)

  • Lee, Won-Jun
    • Journal of Life Science
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    • v.17 no.7 s.87
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    • pp.976-982
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    • 2007
  • The purpose of this study was to determine the modulating effects of histone deacetylase inhibitor, trichostatin A, on the differentiation of mouse C2C12 myoblasts. We demonstrated that trichostatin A induced morphological changes of C2C12 myoblasts into smooth muscles and significantly increased the gene expression of smooth muscle markers including smooth muscle ${\alpha}-actin$ and transgelin. These results were due to the change in the expression level of cell cycle regulators in trichostatin A-treated C2C12 cells. Real-time PCR data revealed that cyclin dependent kinase inhibitor, p21, mRNA expression was significantly increased in trichostatin A-treated C2C12 cells. However, trichostaDn A rapidly decreased cyclin Dl mRNA expression necessary for cell cycle progression in 24hr after treatment. In conclusion, the strong inhibitory effects of trichostatin A on histone deacetylation induced transdifferentiation of C2C12 myoblasts into smooth muscle cells and these results are partly due to the changes in the expression of cell cycle regulators such as p21 and cyclin D1.

Effects of dietary polyphenol (-)-epigallocatechin-3-gallate on the differentiation of mouse C2C12 myoblasts (식이성 폴리페놀 (-)-epigallocatechin-3-gallate가 mouse C2C12 myoblast 분화에 미치는 영향)

  • Kim, Hye-Jin;Lee, Won-Jun
    • Journal of Life Science
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    • v.17 no.3 s.83
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    • pp.420-426
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    • 2007
  • In the present investigation, we studied the modulating effects of (-)-epigallocatechin-3-gallate(EGCG) on the differentiation of mouse C2C12 myoblasts. We found that the strong inhibitory effect of EGCG on DNA methyltransferase-mediated DNA methylation induced transdifferentiation of C2C12 myoblasts into smooth muscle cells demonstrated by both morphological changes and immunofluorescent staining. C2C12 myoblasts treated with EGCG for 4 days expressed smooth muscle ${\alpha}-actin$ protein. Real-time PCR data revealed that smooth muscle ${\alpha}-actin$ mRNA was induced by EGCG treated C2C12 myoblasts in a concentration-dependent manner. Smooth muscle ${\alpha}-actin$ mRNA concentration increased 330% and 490% after 2 and 3 days of 50 ${\mu}M$ of EGCG treatment. The expression of another smooth muscle marker, transgelin, mRNA was also increased up to 9-fold by 4 days of EGCG treatment compared with control in a concentration-dependent manner. These results suggested that C2C12 enables to transdifferentiate into smooth muscle when gene expression patterns are changed by the inhibition of DNA methylation induced by EGCG. In conclusion, transdifferentiation of C2C12 myoblasts into smooth muscle is resulted from the modulating effects of EGCG on DNA methylation which subsequently results in changing the expression pattern of several genes playing a critical role in the differentiation of C2C12 myoblasts.

2, 4-Thiazolidindion Induced Plasticity of Myoblast (C2C12) and Satellite Cells (Porcine) - A Comparative Study

  • Singh, N.K.;Chae, H.S.;Hwang, I.H.;Yoo, Y.M.;Ahn, C.N.;Lee, H.J.;Park, H.J.;Chung, H.Y.
    • Asian-Australasian Journal of Animal Sciences
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    • v.20 no.7
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    • pp.1115-1119
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    • 2007
  • This study was conducted to determine the difference between satellite cells (porcine) and myoblasts (C2C12) in their differentiation under the influence of 2, 4-thiazolidindion. C2C12 myoblast cells and porcine satellite cells (isolated from 10 d old $Landrace{\times}Duroc$ piglets) were grown to absolute confluency. Post confluent cells (day 0) were further exposed to adipogenic induction medium along with 2, 4-thiazolidindion ($8{\mu}M$) for 2 d. Thereafter, cells were exposed to 2, 4-thiazolidindion alone every 2 d till day 10 and analysed. The control was cultured in differentiation medium without any treatment. Increased (p<0.05) expression of transcriptional factors i.e. C/EBP-${\alpha}$ and PPAR-${\gamma}$ and transition of cells to adipocyte morphology was noticed from 2 d and 4 d onwards in satellite cells (Porcine) and myoblasts (C2C12) respectively. Myogenesis was observed to be suppressed completely in case of satellite cells compared to myoblasts in response to 2, 4-thiazolidindion. Pax-7 (transcriptional factor) appeared as a sole entity to satellite cells only, as it was not identified in case of myoblasts. Although both the cells were converting to adipoblasts, the degree of their conversion was different in response to 2, 4-thiazolidindion. Therefore, the hypothesis that satellite cells contribute various domains to the growing myoblasts appeared obscured and found to be dependent on the proliferative energy/or degree of fusion. However, it revealed satellite cells as currency to myoblasts/muscle.

Effect of Differentiation for Mouse Myoblast $C_{2}C_{12}$ Cells against Myostatin expression from Dodamtang (도담탕(導痰湯)이 $C_{2}C_{12}$세포주로부터 myostatin발현에 의한 심근에 미치는 영향)

  • Lee, You-Seung;Shin, Yoo-Jeong;Park, Jong-Hyuk;Kim, Seung-Mo;Paek, Kyung-Min;Park, Chi-Sang
    • The Journal of Internal Korean Medicine
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    • v.29 no.1
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    • pp.243-257
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    • 2008
  • Myostatin, a negative regulator of myogenesis, is shown to function by controlling the proliferation of myoblasts. In this study we show that myostatin is an inhibitor of myoblast differentiation and that this inhibition is mediated through Smad 3. To determine MyoD expression by Dodamtang treatment, we compared the expression pattern of $C_{2}C_{12}$ mouse myoblasts that constitutively express myostatin with control cells. In vitro, increasing concentrations of Dodamtang reversibly prevented the myogenic blockage of myoblasts by myostatin expression. ELISA assay, Western and confocal analysis indicated that treatment of Dodamtang to the low serum culture media increased the levels of MyoD leading to the inhibition of myogenic differentiation by myostatin. The stable transfection of $C_{2}C_{12}$ myoblasts with myostatin expressing constructs did rescue MyoD-induced myogenic differentiation. Consistent with this, the treatment of Dodamtang rescued the expression of a MyoD in $C_{2}C_{12}$ myoblasts treated with myostatin. Taken together, these results suggest that induction of MyoD by Dodamtang inhibits myostatin activity and expression via SMAD3 resulting in the rescue of the myoblasts to differentiate into myotubes. Thus we propose that myostatin action by Dodamtang plays a critical role in myogenic differentiation and that the muscular hyperplasia and hypertrophy seen in animals that blockage of functional myostatin is because of deregulated proliferation and differentiation of myoblasts.

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Antioxidant Activities and Protective Effects of Hot Water Extract from Curcuma longa L. on Oxidative Stress-Induced C2C12 Myoblasts (강황 열수 추출물의 항산화 활성 및 C2C12 Myoblasts의 산화적 손상에 대한 보호 효과)

  • Jeong, Hye-Jin;Kim, Shintae;Park, Jeongjin;Kim, Ki Hong;Kim, Kyungmi;Jun, Woojin
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.46 no.11
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    • pp.1408-1413
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    • 2017
  • The aim of this study was to investigate the antioxidant activities and protective effects of hot water extract from Curcuma longa L. (CLW) on oxidative stress-induced C2C12 myoblasts. Antioxidant activities of CLW were evaluated based on 1,1-diphenyl-2-picrylhydrazyl (DPPH) and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical scavenging activities. Protective effects of CLW on oxidative stress-induced C2C12 myoblasts were determined based on cytotoxicity, $H_2O_2$ protective activity, and intracellular reactive oxygen species (ROS) level. DPPH and ABTS radical scavenging activities represented by $SC_{50}$ were $188.5{\pm}3.0{\mu}g/mL$ and $92.0{\pm}0.9{\mu}g/mL$, respectively. Using C2C12 myoblasts, CLW treatment increased cell viability against oxidative stress-induced cell death. Further, CLW treatment reduced the intracellular ROS level in cells treated with $H_2O_2$. These results suggest that CLW might have the capability to protect oxidative stress-induced C2C12 myoblasts.

Studies on the Differentiation of Skeletal Muscle Cells in vitro:Protein Kinase C in the Differentiation of Skeletal Muscle Cells (근세포 분화에 관한 연구 : 근세포 분화에 있어서 Protein Kinase C)

  • 최원철;김한도;김정락
    • The Korean Journal of Zoology
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    • v.34 no.2
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    • pp.131-141
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    • 1991
  • Treating 12-O-tetradecanoyIphorboI 13-acetate -TPA) or platelet~derived growth factor(PDGF), the signal transduction of protein Idnase C (PKC) is occurred by the phosphoryladon. However the targeting proteins phosphorylated by PKC were found to be different proteins in molecular weights when WA or PDGF wa~ treated to the myoblast. In the WA-treated myoblast cells, the protein of Mr. 20 I(d was phosphorylated. In the PDGF-treated cells, the protein of Mr. 40 Kd was phosphrylated, while the protein of Mr. 20 Kd which phosphorylated in the WA-treatment was dephosphorylated. These results indicate that not only WA and PDGF &e different in activating the signal transduction pathways, but also they may involve in the down reguladon of PI(C during the long-term treatment But PDGF gave rise more rapidly down reguladon than in the case of WA. Using immunocytochemical approach, two disdnct PKC isozymes, PKC II and PKC III, have been localized in cytoplasm and both cytoplasm and nuclsolus, respectively. Ther'efore, the expression of two types of PKC in the myoblast suggests that the isozymes of PKC may involve in each different pathway of signal transduction or down-reguladon.

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