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- W564544270 abstract "Our study is focused on the importance that NGF could have in the molecular mechanisms that regulate the muscular system analyzing its effective role on growth, survival and differentiation of myogenic cells. We performed an in vitro protocol utilizing L6C5 rat skeletal myoblasts, already characterized by our group. L6C5 myoblasts and myotubes were exposed to different concentrations of exogenous NGF (20ng/ml) and were then collected at different time during differentiation to be analyzed for cell growth and survival (MTS assay) the expression of specific myogenic markers (Myogenin and MHC), fusion rate (immunocytoquimical analysis), NFkB activation (Elisa assay), and induction of apoptosis (nuclear fragmentation, Tunel). Studying the influence of the neurotrophic factor on cell growth, we verified that NGF is able to sustain myoblast survival in FBS-free medium, but it cannot induce changes in the rate of cell growth. As regards the effective role on differentiation we demonstrated that NGF (20ng/ml) determines an increase in the differentiative potential of this cell line inducing an over-expression of myogenin in the first phases of the process (2 days in DM), with a consequent hypertrophy of fibers due to an increase in the fusion rate. On the contrary the MHC expression, a terminal differentiation marker, seems not be modulated by NGF. At the same differentiation step (2 days from DM), we found that NGF (20ng/ml) modulates the activity of NFkB, possibly leading to decrease of L6C5 cell susceptibility to apoptosis induced by oxidative stress. When L6C5 myoblasts were maintained in the proliferation medium (GM), NGF can improve the L6C5 myoblast survival towards oxidative stress, but without modifying their susceptibility to apoptosis, or the binding activity of NFkB. Since the p75 low affinity receptor resulted the only one expressed in L6C5 myoblasts and myotubes, we blocked its endogenous action trough an anti-p75 mAb, but this inhibition did not cause a significant increase of apoptotic nuclei, suggesting that the transduction pathway activated by p75 is not the main process involved in the susceptibility of L6C5 myoblasts to apoptotic death. Indeed, although L6C5 cells did not express TrkA receptor, our cellular model is sensitive to treatments with a specific TRK receptor’s inhibitor (AG879), which caused a significant decrease in cell survival together with a reduction in Akt phosphorylation and apoptotic proteolysis of poli (ADP ribose) polymerase enzyme (PARP)." @default.
- W564544270 created "2016-06-24" @default.
- W564544270 creator A5049602623 @default.
- W564544270 date "2008-05-28" @default.
- W564544270 modified "2023-09-27" @default.
- W564544270 title "NGF e regolazione del sistema muscolare: effetto sul differenziamento e sulla sopravvivenza delle cellule miogeniche" @default.
- W564544270 hasPublicationYear "2008" @default.
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