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Expressed sequence tags for bovine muscle satellite cells, myotube formed-cells and adipocyte-like cells.

Lee EJ, Kamli MR, Pokharel S, Malik A, Tareq KM, Roouf Bhat A, Park HB, Lee YS, Kim S, Yang B, Chung KY, Choi I - PLoS ONE (2013)

Bottom Line: 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.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.

View Article: PubMed Central - PubMed

Affiliation: School of Biotechnology, Yeungnam University, Gyeongsan, Republic of Korea ; Bovine Genome Resources Bank, Yeungnam University, Gyeongsan, Republic of Korea.

ABSTRACT

Background: Muscle 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.

Results: A 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.

Conclusion: In 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.

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Related in: MedlinePlus

Biochemical pathway analysis of highly expressed gene list.Representative KEGG pathway for A) MSC, B) MFC and C) ALC.
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pone-0079780-g003: Biochemical pathway analysis of highly expressed gene list.Representative KEGG pathway for A) MSC, B) MFC and C) ALC.

Mentions: We also identified the biochemical pathways of MFC258, MSC233 and ALC248 genes annotated in the present study. FASTA formatted amino acid sequences of DAVID annotated genes in these sets were fed into the KAAS for prediction of various pathways. A total of 130 pathways were predicted for MFC258, whereas 114 and 128 pathways were predicted for MSC233 and ALC248, respectively. A representative pathway for each of the three categories is shown in Figure 3 and a complete list of all pathways is provided in Table S4. Proteins involved in various stages of cellular differentiation pathways including proteoglycans in cancer, regulation of actin cytoskeleton, focal adhesion, tight junction, ribosome, oxidative phosphorylation, ECM-receptor interaction, and various important signaling pathways including the MAPK signaling, Wnt signaling, Hippo signaling, TGF-beta signaling, PI3K-Akt signaling and calcium signaling pathways were represented by unigenes derived from our EST dataset. These data provide substantial evidence that the ESTs generated in this study offer an important resource for MSC differentiation related gene discovery and future functional studies.


Expressed sequence tags for bovine muscle satellite cells, myotube formed-cells and adipocyte-like cells.

Lee EJ, Kamli MR, Pokharel S, Malik A, Tareq KM, Roouf Bhat A, Park HB, Lee YS, Kim S, Yang B, Chung KY, Choi I - PLoS ONE (2013)

Biochemical pathway analysis of highly expressed gene list.Representative KEGG pathway for A) MSC, B) MFC and C) ALC.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3818215&req=5

pone-0079780-g003: Biochemical pathway analysis of highly expressed gene list.Representative KEGG pathway for A) MSC, B) MFC and C) ALC.
Mentions: We also identified the biochemical pathways of MFC258, MSC233 and ALC248 genes annotated in the present study. FASTA formatted amino acid sequences of DAVID annotated genes in these sets were fed into the KAAS for prediction of various pathways. A total of 130 pathways were predicted for MFC258, whereas 114 and 128 pathways were predicted for MSC233 and ALC248, respectively. A representative pathway for each of the three categories is shown in Figure 3 and a complete list of all pathways is provided in Table S4. Proteins involved in various stages of cellular differentiation pathways including proteoglycans in cancer, regulation of actin cytoskeleton, focal adhesion, tight junction, ribosome, oxidative phosphorylation, ECM-receptor interaction, and various important signaling pathways including the MAPK signaling, Wnt signaling, Hippo signaling, TGF-beta signaling, PI3K-Akt signaling and calcium signaling pathways were represented by unigenes derived from our EST dataset. These data provide substantial evidence that the ESTs generated in this study offer an important resource for MSC differentiation related gene discovery and future functional studies.

Bottom Line: 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.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.

View Article: PubMed Central - PubMed

Affiliation: School of Biotechnology, Yeungnam University, Gyeongsan, Republic of Korea ; Bovine Genome Resources Bank, Yeungnam University, Gyeongsan, Republic of Korea.

ABSTRACT

Background: Muscle 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.

Results: A 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.

Conclusion: In 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.

Show MeSH
Related in: MedlinePlus