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Further characterisation of the molecular signature of quiescent and activated mouse muscle satellite cells.

Gnocchi VF, White RB, Ono Y, Ellis JA, Zammit PS - PLoS ONE (2009)

Bottom Line: To date though, there is a paucity of native markers that can be used to easily identify quiescent satellite cells, with Pax7 probably being the best that is currently available.Conversely, Jagged-1, a Notch ligand, was not expressed in quiescent satellite cells but was induced upon activation.These findings further contribute to defining the molecular signature of muscle satellite cells.

View Article: PubMed Central - PubMed

Affiliation: Randall Division of Cell and Molecular Biophysics, King's College London, Guy's Campus, London, UK.

ABSTRACT
Satellite cells are the resident stem cells of adult skeletal muscle. To date though, there is a paucity of native markers that can be used to easily identify quiescent satellite cells, with Pax7 probably being the best that is currently available. Here we have further characterized a number of recently described satellite cell markers, and also describe novel ones. Caveolin-1, integrin alpha7 and the calcitonin receptor proved reliable markers for quiescent satellite cells, being expressed by all satellite cells identified with Pax7. These three markers remained expressed as satellite cells were activated and underwent proliferation. The nuclear envelope proteins lamin A/C and emerin, mutations in which underlie Emery-Dreifuss muscular dystrophy, were also expressed in both quiescent and proliferating satellite cells. Conversely, Jagged-1, a Notch ligand, was not expressed in quiescent satellite cells but was induced upon activation. These findings further contribute to defining the molecular signature of muscle satellite cells.

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Related in: MedlinePlus

Marker protein levels reflect gene expression in quiescent and activated satellite cells.Total RNA was obtained from quiescent (T0) and activated (T48) satellite cells and transcript-specific primers used to semi-quantitatively amplify the mRNA of markers. Amplification of Gapdh transcript was used as control of the RNA content. Pax7, MyoD and myogenin transcripts levels were included as control of the activation progression. While caveolin-1 and CTR transcripts appeared to be more abundant in satellite cells at T0, mRNA for integrin α7, Emd and Lmna were unaltered. Jagged-1 and MyoD transcript were increased in activated satellite cells, reflecting the protein levels. Myogenin transcript, as expected, was absent in quiescent satellite cells and appeared after 48 hrs in culture. NTC is no-template control.
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pone-0005205-g004: Marker protein levels reflect gene expression in quiescent and activated satellite cells.Total RNA was obtained from quiescent (T0) and activated (T48) satellite cells and transcript-specific primers used to semi-quantitatively amplify the mRNA of markers. Amplification of Gapdh transcript was used as control of the RNA content. Pax7, MyoD and myogenin transcripts levels were included as control of the activation progression. While caveolin-1 and CTR transcripts appeared to be more abundant in satellite cells at T0, mRNA for integrin α7, Emd and Lmna were unaltered. Jagged-1 and MyoD transcript were increased in activated satellite cells, reflecting the protein levels. Myogenin transcript, as expected, was absent in quiescent satellite cells and appeared after 48 hrs in culture. NTC is no-template control.

Mentions: Having established the dynamics of the different marker proteins in quiescent and activated satellite cells using immunocytochemistry, we wished to compare the mRNA expression profiles of these genes, so we performed semi-quantitative RT-PCR of their transcripts. We collected quiescent satellite cells (T0), and activated satellite cells after 48 hrs in culture (T48) from isolated myofibres, and purified the RNA. Amplifying the transcripts of the different markers we found that the mRNA levels were in accordance with the protein levels observed using immunostaining (Figure 4). Caveolin-1, integrin α7, CTR, Emd and Lmna transcripts were present in both quiescent and activated satellite cells, with a slightly higher amount in quiescent satellite cells compared to activated ones for caveolin-1 and CTR. Interestingly, the Jagged-1 transcript was already present at T0 - although we could not detect Jagged-1 protein by immunocytochemistry - but was more abundant in activated satellite cells. As expected, the mRNA levels of MyoD increased upon activation.


Further characterisation of the molecular signature of quiescent and activated mouse muscle satellite cells.

Gnocchi VF, White RB, Ono Y, Ellis JA, Zammit PS - PLoS ONE (2009)

Marker protein levels reflect gene expression in quiescent and activated satellite cells.Total RNA was obtained from quiescent (T0) and activated (T48) satellite cells and transcript-specific primers used to semi-quantitatively amplify the mRNA of markers. Amplification of Gapdh transcript was used as control of the RNA content. Pax7, MyoD and myogenin transcripts levels were included as control of the activation progression. While caveolin-1 and CTR transcripts appeared to be more abundant in satellite cells at T0, mRNA for integrin α7, Emd and Lmna were unaltered. Jagged-1 and MyoD transcript were increased in activated satellite cells, reflecting the protein levels. Myogenin transcript, as expected, was absent in quiescent satellite cells and appeared after 48 hrs in culture. NTC is no-template control.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2666265&req=5

pone-0005205-g004: Marker protein levels reflect gene expression in quiescent and activated satellite cells.Total RNA was obtained from quiescent (T0) and activated (T48) satellite cells and transcript-specific primers used to semi-quantitatively amplify the mRNA of markers. Amplification of Gapdh transcript was used as control of the RNA content. Pax7, MyoD and myogenin transcripts levels were included as control of the activation progression. While caveolin-1 and CTR transcripts appeared to be more abundant in satellite cells at T0, mRNA for integrin α7, Emd and Lmna were unaltered. Jagged-1 and MyoD transcript were increased in activated satellite cells, reflecting the protein levels. Myogenin transcript, as expected, was absent in quiescent satellite cells and appeared after 48 hrs in culture. NTC is no-template control.
Mentions: Having established the dynamics of the different marker proteins in quiescent and activated satellite cells using immunocytochemistry, we wished to compare the mRNA expression profiles of these genes, so we performed semi-quantitative RT-PCR of their transcripts. We collected quiescent satellite cells (T0), and activated satellite cells after 48 hrs in culture (T48) from isolated myofibres, and purified the RNA. Amplifying the transcripts of the different markers we found that the mRNA levels were in accordance with the protein levels observed using immunostaining (Figure 4). Caveolin-1, integrin α7, CTR, Emd and Lmna transcripts were present in both quiescent and activated satellite cells, with a slightly higher amount in quiescent satellite cells compared to activated ones for caveolin-1 and CTR. Interestingly, the Jagged-1 transcript was already present at T0 - although we could not detect Jagged-1 protein by immunocytochemistry - but was more abundant in activated satellite cells. As expected, the mRNA levels of MyoD increased upon activation.

Bottom Line: To date though, there is a paucity of native markers that can be used to easily identify quiescent satellite cells, with Pax7 probably being the best that is currently available.Conversely, Jagged-1, a Notch ligand, was not expressed in quiescent satellite cells but was induced upon activation.These findings further contribute to defining the molecular signature of muscle satellite cells.

View Article: PubMed Central - PubMed

Affiliation: Randall Division of Cell and Molecular Biophysics, King's College London, Guy's Campus, London, UK.

ABSTRACT
Satellite cells are the resident stem cells of adult skeletal muscle. To date though, there is a paucity of native markers that can be used to easily identify quiescent satellite cells, with Pax7 probably being the best that is currently available. Here we have further characterized a number of recently described satellite cell markers, and also describe novel ones. Caveolin-1, integrin alpha7 and the calcitonin receptor proved reliable markers for quiescent satellite cells, being expressed by all satellite cells identified with Pax7. These three markers remained expressed as satellite cells were activated and underwent proliferation. The nuclear envelope proteins lamin A/C and emerin, mutations in which underlie Emery-Dreifuss muscular dystrophy, were also expressed in both quiescent and proliferating satellite cells. Conversely, Jagged-1, a Notch ligand, was not expressed in quiescent satellite cells but was induced upon activation. These findings further contribute to defining the molecular signature of muscle satellite cells.

Show MeSH
Related in: MedlinePlus