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Altered functional differentiation of mesoangioblasts in a genetic myopathy.

Altomare C, Barile L, Rocchetti M, Sala L, Crippa S, Sampaolesi M, Zaza A - J. Cell. Mol. Med. (2013)

Bottom Line: To this end, cMabs were compared with neonatal cardiomyocytes (CMs) and skeletal muscle C2C12 cells, representative of cardiac and skeletal muscle respectively.Contrary to previous reports, wild-type cMabs failed to show functional differentiation towards either cell type.We conclude that miRNA669a expression, ablated by βSG deletion, may prevent functional differentiation of cMabs towards the skeletal muscle phenotype.

View Article: PubMed Central - PubMed

Affiliation: Department of Biotechnologies and Biosciences, University of Milano-Bicocca, Milan, Italy.

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L-type Ca2+ current (ICaL) analysis and channel expression. (a) Representative ICaL recordings in βSG−/−, C2C12 myotubes and CMs (voltage protocol in top panel); (b) V dependency of ICaL inactivation time constants (τinact); (c) Peak ICaL I/V curves (top) and (d) steady-state activation curves (Boltzman fitting). (e) Expression (RT-PCR) of skeletal (Cav1.1) and cardiac (Cav 1.2) Ca2+ channel isoforms (GAPDH as reference). Open circles: C2C12 (N = 9); filled squares: βSG−/− (N = 6); filled triangles: CMs (N = 5). (f) Confocal immunofluorescence analysis of Cav 1.1 (skeletal) and cardiac Cav 1.2 (cardiac) isoforms in βSG−/− myotubes. Signals for Cav (green) and nuclei (blue-Hoechst) are superimposed on the light transmission image.
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fig05: L-type Ca2+ current (ICaL) analysis and channel expression. (a) Representative ICaL recordings in βSG−/−, C2C12 myotubes and CMs (voltage protocol in top panel); (b) V dependency of ICaL inactivation time constants (τinact); (c) Peak ICaL I/V curves (top) and (d) steady-state activation curves (Boltzman fitting). (e) Expression (RT-PCR) of skeletal (Cav1.1) and cardiac (Cav 1.2) Ca2+ channel isoforms (GAPDH as reference). Open circles: C2C12 (N = 9); filled squares: βSG−/− (N = 6); filled triangles: CMs (N = 5). (f) Confocal immunofluorescence analysis of Cav 1.1 (skeletal) and cardiac Cav 1.2 (cardiac) isoforms in βSG−/− myotubes. Signals for Cav (green) and nuclei (blue-Hoechst) are superimposed on the light transmission image.

Mentions: As already evident by gross examination of sample recordings (Fig. 5a), inactivation kinetics was similar in βSG−/− and C2C12 myotubes and differed from that of CMs (also notice the difference in the timescale). Figure 5b shows that inactivation time constants (τinact) had a shallow dependency on membrane potential in all cell types, but were distinctly shorter in CMs (n = 9) than in βSG−/− or C2C12 myotubes (P < 0.05 at all potentials, n = 7 and n = 12 respectively).


Altered functional differentiation of mesoangioblasts in a genetic myopathy.

Altomare C, Barile L, Rocchetti M, Sala L, Crippa S, Sampaolesi M, Zaza A - J. Cell. Mol. Med. (2013)

L-type Ca2+ current (ICaL) analysis and channel expression. (a) Representative ICaL recordings in βSG−/−, C2C12 myotubes and CMs (voltage protocol in top panel); (b) V dependency of ICaL inactivation time constants (τinact); (c) Peak ICaL I/V curves (top) and (d) steady-state activation curves (Boltzman fitting). (e) Expression (RT-PCR) of skeletal (Cav1.1) and cardiac (Cav 1.2) Ca2+ channel isoforms (GAPDH as reference). Open circles: C2C12 (N = 9); filled squares: βSG−/− (N = 6); filled triangles: CMs (N = 5). (f) Confocal immunofluorescence analysis of Cav 1.1 (skeletal) and cardiac Cav 1.2 (cardiac) isoforms in βSG−/− myotubes. Signals for Cav (green) and nuclei (blue-Hoechst) are superimposed on the light transmission image.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3823023&req=5

fig05: L-type Ca2+ current (ICaL) analysis and channel expression. (a) Representative ICaL recordings in βSG−/−, C2C12 myotubes and CMs (voltage protocol in top panel); (b) V dependency of ICaL inactivation time constants (τinact); (c) Peak ICaL I/V curves (top) and (d) steady-state activation curves (Boltzman fitting). (e) Expression (RT-PCR) of skeletal (Cav1.1) and cardiac (Cav 1.2) Ca2+ channel isoforms (GAPDH as reference). Open circles: C2C12 (N = 9); filled squares: βSG−/− (N = 6); filled triangles: CMs (N = 5). (f) Confocal immunofluorescence analysis of Cav 1.1 (skeletal) and cardiac Cav 1.2 (cardiac) isoforms in βSG−/− myotubes. Signals for Cav (green) and nuclei (blue-Hoechst) are superimposed on the light transmission image.
Mentions: As already evident by gross examination of sample recordings (Fig. 5a), inactivation kinetics was similar in βSG−/− and C2C12 myotubes and differed from that of CMs (also notice the difference in the timescale). Figure 5b shows that inactivation time constants (τinact) had a shallow dependency on membrane potential in all cell types, but were distinctly shorter in CMs (n = 9) than in βSG−/− or C2C12 myotubes (P < 0.05 at all potentials, n = 7 and n = 12 respectively).

Bottom Line: To this end, cMabs were compared with neonatal cardiomyocytes (CMs) and skeletal muscle C2C12 cells, representative of cardiac and skeletal muscle respectively.Contrary to previous reports, wild-type cMabs failed to show functional differentiation towards either cell type.We conclude that miRNA669a expression, ablated by βSG deletion, may prevent functional differentiation of cMabs towards the skeletal muscle phenotype.

View Article: PubMed Central - PubMed

Affiliation: Department of Biotechnologies and Biosciences, University of Milano-Bicocca, Milan, Italy.

Show MeSH
Related in: MedlinePlus