Expressed Sequence Tags for Bovine Muscle Satellite Cells, Myotube Formed-Cells and Adipocyte-Like Cells

Abstract

BackgroundMuscle satellite cells (MSCs) represent a devoted stem cell population that is responsible for postnatal muscle growth and skeletal muscle regeneration. An important characteristic of MSCs is that they encompass multi potential mesenchymal stem cell activity and are able to differentiate into myocytes and adipocytes. To achieve a global view of the genes differentially expressed in MSCs, myotube formed-cells (MFCs) and adipocyte-like cells (ALCs), we performed large-scale EST sequencing of normalized cDNA libraries developed from bovine MSCs.ResultsA total of 24,192 clones were assembled into 3,333 clusters, 5,517 singletons and 3,842contigs. Functional annotation of these unigenes revealed that a large portion of the differentially expressed genes are involved in cellular and signaling processes. Database for Annotation, Visualization and Integrated Discovery (DAVID) functional analysis of three subsets of highly expressed gene lists (MSC233, MFC258, and ALC248) highlighted some common and unique biological processes among MSC, MFC and ALC. Additionally, genes that may be specific to MSC, MFC and ALC are reported here, and the role of dimethylarginine dimethylaminohydrolase2 (DDAH2) during myogenesis and hemoglobin subunit alpha2 (HBA2) during transdifferentiation in C2C12 were assayed as a case study. DDAH2 was up-regulated during myognesis and knockdown of DDAH2 by siRNA significantly decreased myogenin (MYOG) expression corresponding with the slight change in cell morphology. In contrast, HBA2 was up-regulated during ALC formation and resulted in decreased intracellular lipid accumulation and CD36 mRNA expression upon knockdown assay.ConclusionIn this study, a large number of EST sequences were generated from the MSC, MFC and ALC. Overall, the collection of ESTs generated in this study provides a starting point for the identification of novel genes involved in MFC and ALC formation, which in turn offers a fundamental resource to enable better understanding of the mechanism of muscle differentiation and transdifferentiation.

Genome-wide DNA Methylation Profiles and Their Relationships with mRNA and the microRNA Transcriptome in Bovine Muscle Tissue (Bos taurine)

“…The methylation of these genes might partially contribute to the bovine growth difference. On the other hand, some well-known genes related to the normal growth and development (EEF1A2, TUBA1B and DUSP1) were observed 70 71 72 . We believed that the methylation of these genes might partially contribute to the bovine growth difference between the fetal and adult stage.…”

Fibromodulin and regulation of the intricate balance between myoblast differentiation to myocytes or adipocyte‐like cells

“…Oil Red O staining was performed as previously described (20). Briefly, cells were fixed with 10% formaldehyde (Sigma‐Aldrich) for 20 min and incubated for 1 h with Oil Red O solution (6:4 dilution of stock: 3.5 mg/ml Oil Red O powder in 100% isopropanol).…”

“…The involvement of FMOD in different cellular processes (14) made us study its regulatory function on MSC fate in the myogenic program. Because MSCs have the ability to differentiate into muscle cells or transdifferentiate into adipocyte‐like cells (20, 21), substantial expression of FMOD in muscle tissue led us to explore the possibility of FMOD involvement in shaping the fate of MSC at the differentiation stage to the myogenic or lipid accumulation paths.…”

Understanding Calcium-Dependent Conformational Changes in S100A1 Protein: A Combination of Molecular Dynamics and Gene Expression Study in Skeletal Muscle

“…In previous studies, our group and other researchers have explored the importance of Ca 2+ signaling and that of the associated proteins in skeletal muscle functions [20][21][22]. The contractile properties of skeletal muscles are largely reliant on the protein expression involved in the signaling of Ca 2+ as this is one of the main signaling and regulatory molecules of myofibers [20].…”