Estimation of anti-diabetic property of Holarrhena pubescens by In vitro method using l6 rat skeletal muscle cell lines

2020 ◽  
Vol 13 (12) ◽  
pp. 6238-6242
Author(s):  
Bonagiri Sreedevi ◽  
Vijaya Kuchana ◽  
S. Shobharani
Keyword(s):  
Skeletal Muscle ◽  
Cell Lines ◽  
Muscle Cell ◽  
Skeletal Muscle Cell ◽  
Rat Skeletal Muscle ◽  
In Vitro Method

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.

scholarly journals Human and rat skeletal muscle single-nuclei multi-omic integrative analyses nominate causal cell types, regulatory elements, and SNPs for complex traits

2021 ◽  
Author(s):  
Peter Orchard ◽  
Nandini Manickam ◽  
Christa Ventresca ◽  
Swarooparani Vadlamudi ◽  
Arushi Varshney ◽  
...  
Keyword(s):  
Skeletal Muscle ◽  
Muscle Cell ◽  
Muscle Fiber ◽  
Cell Types ◽  
Chromatin Accessibility ◽  
Skeletal Muscle Cell ◽  
Rna Seq ◽  
Rat Skeletal Muscle ◽  
Integrative Analyses ◽  
Single Nucleus

Skeletal muscle accounts for the largest proportion of human body mass, on average, and is a key tissue in complex diseases and mobility. It is composed of several different cell and muscle fiber types. Here, we optimize single-nucleus ATAC-seq (snATAC-seq) to map skeletal muscle cell–specific chromatin accessibility landscapes in frozen human and rat samples, and single-nucleus RNA-seq (snRNA-seq) to map cell-specific transcriptomes in human. We additionally perform multi-omics profiling (gene expression and chromatin accessibility) on human and rat muscle samples. We capture type I and type II muscle fiber signatures, which are generally missed by existing single-cell RNA-seq methods. We perform cross-modality and cross-species integrative analyses on 33,862 nuclei and identify seven cell types ranging in abundance from 59.6% to 1.0% of all nuclei. We introduce a regression-based approach to infer cell types by comparing transcription start site–distal ATAC-seq peaks to reference enhancer maps and show consistency with RNA-based marker gene cell type assignments. We find heterogeneity in enrichment of genetic variants linked to complex phenotypes from the UK Biobank and diabetes genome-wide association studies in cell-specific ATAC-seq peaks, with the most striking enrichment patterns in muscle mesenchymal stem cells (∼3.5% of nuclei). Finally, we overlay these chromatin accessibility maps on GWAS data to nominate causal cell types, SNPs, transcription factor motifs, and target genes for type 2 diabetes signals. These chromatin accessibility profiles for human and rat skeletal muscle cell types are a useful resource for nominating causal GWAS SNPs and cell types.

Specific effect of corticoids on acetylcholine receptor expression in rat skeletal muscle cell cultures

1992 ◽  
Vol 31 (2) ◽  
pp. 285-293 ◽  
Author(s):  
J.-T. Vilquin ◽  
S. Braun ◽  
P. Labouret ◽  
G. Zuber ◽  
C. Tranchant ◽  
...  
Keyword(s):  
Skeletal Muscle ◽  
Acetylcholine Receptor ◽  
Muscle Cell ◽  
Cell Cultures ◽  
Specific Effect ◽  
Skeletal Muscle Cell ◽  
Receptor Expression ◽  
Rat Skeletal Muscle

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 A skeletal muscle-specific enhancer regulated by factors binding to E and CArG boxes is present in the promoter of the mouse myosin light-chain 1A gene.

1995 ◽  
Vol 15 (8) ◽  
pp. 4585-4596 ◽  
Author(s):  
F Catala ◽  
R Wanner ◽  
P Barton ◽  
A Cohen ◽  
W Wright ◽  
...  
Keyword(s):  
Skeletal Muscle ◽  
Cell Lines ◽  
Muscle Cell ◽  
Light Chain ◽  
Myosin Light Chain ◽  
Primary Cultures ◽  
Skeletal Muscle Cell ◽  
Proximal Promoter ◽  
Carg Box ◽  
Specific Regulation

The mouse myosin light-chain 1A (MLC1A) gene, expressed in the atria of the adult heart, is one of the first muscle genes to be activated when skeletal as well as cardiac muscles form in the embryo. It is also transcribed in skeletal muscle cell lines at the onset of differentiation. Transient transfection assays of mouse skeletal muscle cell lines with DNA constructs containing MLC1A promoter fragments fused to the chloramphenicol acetyltransferase (CAT) gene show that the first 630 bp of the promoter is sufficient to direct expression of the reporter gene during myotube formation. Two E boxes located at bp -76 and -519 are necessary for this regulation. MyoD and myogenin proteins bind to them as heterodimers with E12 protein and, moreover, transactivate them in cotransfection experiments with the MLC1A promoter in nonmuscle cells. Interestingly, the effect of mutating each E box is less striking in primary cultures than in the C2 or Sol8 muscle cell line. A DNA fragment from bp -36 to -597 confers tissue- and stage-specific activity to the herpes simplex virus thymidine kinase promoter in both orientations, showing that the skeletal muscle-specific regulation of the MLC1A gene is under the control of a muscle-specific enhancer which extends into the proximal promoter region. At bp -89 is a diverged CArG box, CC(A/T)6AG, which binds the serum response factor (SRF) in myotube nuclear extracts, as does the wild-type sequence, CC(A/T)6GG. Both types of CArG box also bind a novel myotube-enriched complex which has contact points with the AT-rich part of the CArG box and adjacent 3' nucleotides. Mutations within the CArG box distinguish between the binding of this complex and binding of SRF; only SRF binding is directly involved in the specific regulation of the MLC1A gene in skeletal muscle cell lines.

Sign in / Sign up

Export Citation Format

Share Document