scholarly journals Cardiac actin is the major actin gene product in skeletal muscle cell differentiation in vitro.

1984 ◽  
Vol 4 (8) ◽  
pp. 1449-1453 ◽  
Author(s):  
W Bains ◽  
P Ponte ◽  
H Blau ◽  
L Kedes
Keyword(s):  
Skeletal Muscle ◽  
Muscle Cell ◽  
Skeletal Muscle Cell ◽  
C2c12 Cells ◽  
Adult Skeletal Muscle ◽  
Mature Adult ◽  
Actin Genes ◽  
Cardiac Actin ◽  
Actin Mrna

We examined the expression of alpha-skeletal, alpha-cardiac, and beta- and gamma-cytoskeletal actin genes in a mouse skeletal muscle cell line (C2C12) during differentiation in vitro. Using isotype-specific cDNA probes, we showed that the alpha-skeletal actin mRNA pool reached only 15% of the level reached in adult skeletal muscle and required several days to attain this peak, which was then stably maintained. However, these cells accumulated a pool of alpha-cardiac actin six times higher than the alpha-skeletal actin mRNA peak within 24 h of the initiation of differentiation. After cells had been cultured for an additional 3 days, this pool declined to 10% of its peak level. In contrast, over 95% of the actin mRNA in adult skeletal muscle coded for alpha-actin. This suggests that C2C12 cells express a pattern of sarcomeric actin genes typical of either muscle development or regeneration and distinct from that seen in mature, adult tissue. Concurrently in the course of differentiation the beta- and gamma-cytoskeletal actin mRNA pools decreased to less than 10% of their levels in proliferating cells. The decreases in beta- and gamma-cytoskeletal actin mRNAs are apparently not coordinately regulated.

Cardiac actin is the major actin gene product in skeletal muscle cell differentiation in vitro

1984 ◽  
Vol 4 (8) ◽  
pp. 1449-1453
Author(s):  
W Bains ◽  
P Ponte ◽  
H Blau ◽  
L Kedes
Keyword(s):  
Skeletal Muscle ◽  
Muscle Cell ◽  
Skeletal Muscle Cell ◽  
C2c12 Cells ◽  
Adult Skeletal Muscle ◽  
Mature Adult ◽  
Actin Genes ◽  
Cardiac Actin ◽  
Actin Mrna

We examined the expression of alpha-skeletal, alpha-cardiac, and beta- and gamma-cytoskeletal actin genes in a mouse skeletal muscle cell line (C2C12) during differentiation in vitro. Using isotype-specific cDNA probes, we showed that the alpha-skeletal actin mRNA pool reached only 15% of the level reached in adult skeletal muscle and required several days to attain this peak, which was then stably maintained. However, these cells accumulated a pool of alpha-cardiac actin six times higher than the alpha-skeletal actin mRNA peak within 24 h of the initiation of differentiation. After cells had been cultured for an additional 3 days, this pool declined to 10% of its peak level. In contrast, over 95% of the actin mRNA in adult skeletal muscle coded for alpha-actin. This suggests that C2C12 cells express a pattern of sarcomeric actin genes typical of either muscle development or regeneration and distinct from that seen in mature, adult tissue. Concurrently in the course of differentiation the beta- and gamma-cytoskeletal actin mRNA pools decreased to less than 10% of their levels in proliferating cells. The decreases in beta- and gamma-cytoskeletal actin mRNAs are apparently not coordinately regulated.

Arachidonic acid supplementation enhances in vitro skeletal muscle cell growth via a COX-2-dependent pathway

2013 ◽  
Vol 304 (1) ◽  
pp. C56-C67 ◽  
Author(s):  
James F. Markworth ◽  
David Cameron-Smith
Keyword(s):  
Skeletal Muscle ◽  
Arachidonic Acid ◽  
Cell Growth ◽  
Muscle Cell ◽  
Myogenic Differentiation ◽  
Skeletal Muscle Cell ◽  
Diverse Range ◽  
Cox 2 ◽  
Dependent Pathway

Arachidonic acid (AA) is the metabolic precursor to a diverse range of downstream bioactive lipid mediators. A positive or negative influence of individual eicosanoid species [e.g., prostaglandins (PGs), leukotrienes, and hydroxyeicosatetraenoic acids] has been implicated in skeletal muscle cell growth and development. The collective role of AA-derived metabolites in physiological states of skeletal muscle growth/atrophy remains unclear. The present study aimed to determine the direct effect of free AA supplementation and subsequent eicosanoid biosynthesis on skeletal myocyte growth in vitro . C2C12 (mouse) skeletal myocytes induced to differentiate with supplemental AA exhibited dose-dependent increases in the size, myonuclear content, and protein accretion of developing myotubes, independent of changes in cell density or the rate/extent of myogenic differentiation. Nonselective (indomethacin) or cyclooxygenase 2 (COX-2)-selective (NS-398) nonsteroidal anti-inflammatory drugs blunted basal myogenesis, an effect that was amplified in the presence of supplemental free AA substrate. The stimulatory effects of AA persisted in preexisting myotubes via a COX-2-dependent (NS-389-sensitive) pathway, specifically implying dependency on downstream PG biosynthesis. AA-stimulated growth was associated with markedly increased secretion of PGF2α and PGE2; however, incubation of myocytes with PG-rich conditioned medium failed to mimic the effects of direct AA supplementation. In vitro AA supplementation stimulates PG release and skeletal muscle cell hypertrophy via a COX-2-dependent pathway.

scholarly journals An in vitro injury model to examine pericyte‐skeletal muscle cell cross talk

2013 ◽  
Vol 27 (S1) ◽  
Author(s):  
Katherine E LaBarbera ◽  
Kevin S O'Fallon ◽  
Priscilla M Clarkson ◽  
Sarah Witkowski
Keyword(s):  
Skeletal Muscle ◽  
Muscle Cell ◽  
Cross Talk ◽  
Skeletal Muscle Cell ◽  
Injury Model

scholarly journals alpha-skeletal and alpha-cardiac actin genes are coexpressed in adult human skeletal muscle and heart.

1983 ◽  
Vol 3 (11) ◽  
pp. 1985-1995 ◽  
Author(s):  
P Gunning ◽  
P Ponte ◽  
H Blau ◽  
L Kedes
Keyword(s):  
Skeletal Muscle ◽  
Smooth Muscle ◽  
Human Skeletal Muscle ◽  
Cdna Clones ◽  
Actin Gene ◽  
Adult Heart ◽  
Adult Human ◽  
Actin Genes ◽  
Cardiac Actin ◽  
Actin Mrna

We determined the actin isotypes encoded by 30 actin cDNA clones previously isolated from an adult human muscle cDNA library. Using 3' untranslated region probes derived from alpha-skeletal, beta- and gamma-actin cDNAs and from an alpha-cardiac actin genomic clone, we showed that 28 of the cDNAs correspond to alpha-skeletal actin transcripts. Unexpectedly, however, the remaining two cDNA clones proved to derive from alpha-cardiac actin mRNA. Sequence analysis confirmed that the two skeletal muscle alpha-cardiac actin cDNAs are derived from transcripts of the cloned alpha-cardiac actin gene. Direct measurements of actin isotype mRNA expression in human skeletal muscle showed that alpha-cardiac actin mRNA is expressed at 5% the level of alpha-skeletal actin. Furthermore, the alpha-cardiac actin gene expressed in skeletal muscle is the same gene which produces alpha-cardiac actin mRNA in the human heart. Of equal surprise, we found that alpha-skeletal actin mRNA accounts for about half of the total actin mRNA in adult heart. Comparison of total actin mRNA levels in adult skeletal muscle and adult heart revealed that the steady-state levels in skeletal muscle are about twofold greater, per microgram of total cellular RNA, than those in heart. Thus, in skeletal muscle and in heart, both of the sarcomeric actin mRNA isotypes are quite abundant transcripts. We conclude that alpha-skeletal and alpha-cardiac actin genes are coexpressed as an actin pair in human adult striated muscles. Since the smooth-muscle actins (aortic and stomach) and the cytoplasmic actins (beta and gamma) are known to be coexpressed in smooth muscle and nonmuscle cells, respectively, we postulate that coexpression of actin pairs may be a common feature of mammalian actin gene expression in all tissues.

scholarly journals Further considerations on in vitro skeletal muscle cell death

2019 ◽  
Vol 03 (04) ◽  
pp. 267 ◽  
Author(s):  
M. Battistelli ◽  
S. Salucci ◽  
S. Burattini ◽  
E. Falcieri
Keyword(s):  
Skeletal Muscle ◽  
Cell Death ◽  
Muscle Cell ◽  
Skeletal Muscle Cell

scholarly journals Designed Functional Dispersion for Insulin Protection from Pepsin Degradation and Skeletal Muscle Cell Proliferation: In Silico and In Vitro Study

Nanomaterials ◽  
2018 ◽  
Vol 8 (10) ◽  
pp. 852 ◽  
Author(s):  
Veera Chittepu ◽  
Poonam Kalhotra ◽  
Tzayhri Gallardo-Velázquez ◽  
Raúl Robles-de la Torre ◽  
Guillermo Osorio-Revilla
Keyword(s):  
Skeletal Muscle ◽  
Cell Proliferation ◽  
Insulin Therapy ◽  
Muscle Cell ◽  
Carbon Nanomaterials ◽  
Pharmaceutical Preparations ◽  
In Vitro Study ◽  
Skeletal Muscle Cell ◽  
Functional Dispersion

Functionalized single-walled carbon nanotubes with polyethylene glycol (PEGylated SWCNTs) are a promising nanomaterial that recently has emerged as the most attractive “cargo” to deliver chemicals, peptides, DNA and RNAs into cells. Insulin therapy is a recommended therapy to treat diabetes mellitus despite its side effects. Recently, functional dispersion made up of bioactive peptides, bioactive compounds and functionalized carbon nanomaterials such as PEGylated SWCNTs have proved to possess promising applications in nanomedicine. In the present study, molecular modeling simulations are utilized to assist in designing insulin hormone-PEGylated SWCNT composites, also called functional dispersion; to achieve this experimentally, an ultrasonication tool was utilized. Enzymatic degradation assay revealed that the designed functional dispersion protects about 70% of free insulin from pepsin. In addition, sulforhodamine B (SRB) assay, the quantification of insulin and glucose levels in differentiated skeletal muscle cell supernatants, reveals that functional dispersion regulates glucose and insulin levels to promote skeletal muscle cell proliferation. These findings offer new perspectives for designed functional dispersion, as potential pharmaceutical preparations to improve insulin therapy and promote skeletal muscle cell health.

scholarly journals Phytochemicals in Chinese chive (Allium tuberosum) Induce the Skeletal Muscle Cell Proliferation via PI3K/Akt/mTOR and Smad Pathways in C2C12 Cells

2021 ◽  
Vol 22 (5) ◽  
pp. 2296
Author(s):  
Mira Oh ◽  
Seo-Young Kim ◽  
SeonJu Park ◽  
Kil-Nam Kim ◽  
Seung Hyun Kim
Keyword(s):  
Skeletal Muscle ◽  
Cell Proliferation ◽  
Muscle Cell ◽  
Muscle Growth ◽  
Skeletal Muscle Cell ◽  
Water Soluble ◽  
C2c12 Cells ◽  
Allium Tuberosum ◽  
Skeletal Muscle Growth ◽  
Chinese Chive

Chinese chive (Allium tuberosum) is a medicinal food that is cultivated and consumed mainly in Asian countries. Its various phytochemicals and physiological effects have been reported, but only a few phytochemicals are available for skeletal muscle cell proliferation. Herein, we isolated a new compound, kaempferol-3-O-(6″-feruloyl)-sophoroside (1), along with one known flavonoid glycoside (2) and six amino acid (3–8) compounds from the water-soluble fraction of the shoot of the Chinese chive. The isolated compounds were identified using extensive spectroscopic methods, including 1D and 2D NMR, and evaluated for their proliferation activity on skeletal muscle cells. Among the tested compounds, newly isolated flavonoid (1) and 5-aminouridine (7) up-regulated PI3K/Akt/mTOR pathways, which implies a positive effect on skeletal muscle growth and differentiation. In particular, compound 1 down-regulated the Smad pathways, which are negative regulators of skeletal muscle growth. Collectively, we suggest that major constituents of Chinese chive, flavonoids and amino acids, might be used in dietary supplements that aid skeletal muscle growth.

scholarly journals Conditionally Immortalised Equine Skeletal Muscle Cell Lines for in Vitro Analysis

2021 ◽  
Author(s):  
Mary Francis Rooney ◽  
Nuno Neto ◽  
Michael Monaghan ◽  
Emmeline Hill ◽  
Richard Porter
Keyword(s):  
Skeletal Muscle ◽  
Cell Lines ◽  
Muscle Cell ◽  
Mitochondrial Function ◽  
Ex Vivo ◽  
Skeletal Muscle Cell ◽  
Cell Phenotype ◽  
Metabolic Function ◽  
Thoroughbred Horse

Abstract BackgroundThoroughbred racehorse performance is largely influenced by a major quantitative trait locus at the myostatin (MSTN) gene which determines aptitude for certain race distances due to a promoter region insertion mutation influencing functional phenotypes in skeletal muscle. To develop an in vitro system for functional experiments we established three novel equine skeletal muscle cell lines reflecting the variation in phenotype associated with MSTN genotype (CC/II, CT/IN and TT/NN for SNP g.66493737C>T/SINE insertion 227 bp polymorphism). Primary equine skeletal muscle myoblasts, isolated from Thoroughbred horse gluteus medius, were conditionally immortalised and evaluated to determine whether cell phenotype and metabolic function were comparable to functional characteristics previously reported for ex vivo skeletal muscle isolated from Thoroughbred horses with each genotype.ResultsPrimary myoblasts conditionally immortalized with the temperature sensitive SV40TtsA58 lentivirus vector successfully proliferated and could revert to their primary cell phenotype and differentiate into multinucleated myotubes. Skeletal muscle fibre type, MSTN gene expression, mitochondrial abundance, and mitochondrial function of the three MSTN genotype cell lines, were consistent with equivalent characterisation of ex vivo skeletal muscle samples with these genotypes. Furthermore, addition of coenzyme Q10 (CoQ10) to the cell lines improved mitochondrial function, an observation consistent with ex vivo skeletal muscle samples with these genotypes following supplementation with CoQ10 in the diet. ConclusionsThe observation that the phenotypic characteristics and metabolic function of the cells lines are equivalent to ex vivo skeletal muscle indicates that this in vitro system will enable efficient and cost-effective analyses of equine skeletal muscle for a range of different applications including understanding metabolic function, testing of nutritional supplements, drug test development and gene doping test development. In the multi-billion-euro international Thoroughbred horse industry research advances in the biological function of skeletal muscle are likely to have considerable impact. Furthermore, this novel genotype-specific system may be adapted and applied to human biomedicine to improve understanding of the effects of myostatin in human physiology and medicine.

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