transcriptomic analysis of proliferative or differentiated myoblasts

Adult skeletal muscle is a plastic tissue that can adapt its size to workload and that can regenerate after demage. Here, we show that RhoA within primary myoblasts is needed for a correct muscle regeneration by controlling satellite cell fusion without affecting their proliferation or their differentiation. Moreover primary cultured myoblast loosing RhoA have no defects in cytoskeleton reorganization however their movement is altered compared to control cells. At the molecular level, we found that RhoA controls the expressions of different genes depending on step of differentiation process, not necessarly already described as implicated in fusion program. These findings unravel the implication of RhoA within satellite cells regulating myoblast fusion in response to demage in order to allow a correct regeneration process. The aim of the analysis is to compare proliferative myoblasts and differentiated myotubes with and wthout RhoA. RhoAfloxed mice were injected or not with tamoxifen in order to delete in vivo RhoA only in satellite cells. Primary myoblast were isolated by RhoAfloxed mice by FACS. The cells isolated by FACS, proliferate and differentiate in culture. RNA was isolated from primary proliferative myoblast and differentiated myotubes isolated by control or RhoA deleted mice. Microarray analysis will help to understand the molecular pathways involved in fusion defect observed in vivo and in vitro. Isolated RNA from 3 independent control and RhoA-lacking cell cultures were used to perform trasncriptomic arrays. Total Rna was isolated from cells at day0, day1 and day3 of differentiation.