Project description:Muscle stem cell quiescence is a complex and dynamic state. In this study we examined the role of the Ubiquitin Protease System in muscle stem cell function and quiescence. Using a genetic mouse model where Nedd4L is selectively deleted in myogenic cells, we characterized the effect of the loss of Nedd4L on muscle stem cells. With RNA-Seq we determined that the genetic deletion of Nedd4L resulted in the transition of the muscle stem cells out of deep quiescence and into Galert. Characterized by the upregulation of cell cycle genes and differentiation genes and loss of expression in numerous known quiescence genes. Nedd4L-cKO muscle stem cells are more prone to differentiation and fail to expand properly in vitro.
Project description:Muscle stem cell quiescence is a complex and dynamic state. In this study we examined the role of the Ubiquitin Protease System in muscle stem cell function and quiescence. Using a genetic mouse model where Nedd4L is selectively deleted in myogenic cells, we characterized the effect of the loss of Nedd4L on muscle stem cells. With RNA-Seq we determined that the genetic deletion of Nedd4L resulted in the transition of the muscle stem cells out of deep quiescence and into Galert. Characterized by the upregulation of cell cycle genes and differentiation genes and loss of expression in numerous known quiescence genes. Nedd4L-cKO muscle stem cells are more prone to differentiation and fail to expand properly in vitro.
Project description:TNFα is a cytokine with multiple biological effects. It activates cascading signal pathways by binding to receptors on the cell membrane, regulating processes such as cell growth, differentiation, apoptosis, and inflammatory responses. The formation of skeletal muscle primarily relies on processes such as myoblasts proliferation, migration, differentiation, cytoskeletal rearrangement, and myotube fusion, which constitute a complex and coordinated process regulated by multiple genes. While research on TNFα has primarily focused on immunology, some studies suggest that TNFα also plays a physiological role in myogenesis, muscle repair, diseases, and atrophy.We isolated the nuclei (N) and cytoplasm (C) of flox and TNFα-CKO primary myoblasts in the early stages of differentiation and attempted to identify new targets in the TNFα signaling pathway.
Project description:Comparison of muscle stem cell preplates and myoblasts. Experiment Overall Design: this experiment include 2 samples and 2 replicates
Project description:Gene expression in satellite cell-derived primary myoblasts islolated from Gata4-loxP mice. Myoblasts were infected with nLacZ for control (Wt), Cre recombinase to knockout GATA4 (KO), or GATA4 expression vector to overexpress GATA4 (OE). Infected myoblasts were cultured in growth medium (day 0) then differentiated into myotubes in differentiation medium for 3 days (day 3).
Project description:Gene expression in satellite cell-derived primary myoblasts islolated from Gata4-loxP mice. Myoblasts were infected with nLacZ for control (Wt), Cre recombinase to knockout GATA4 (KO), or GATA4 expression vector to overexpress GATA4 (OE). Infected myoblasts were cultured in growth medium (day 0) then differentiated into myotubes in differentiation medium for 3 days (day 3). Total 18 samples. Three replicates in each myoblast; GATA4-Wt, -KO, and -OE myoblasts at day 0 and day 3.
Project description:To get insight into the transcriptomic changes caused by HDAC11 loss at early myogenic differentiation, we performed RNA sequencing (RNA-Seq) in 24 h differentiated primary myoblasts from WT and HDAC11-deficient mice. Satellite cell-derived primary myoblasts were obtained after FACS isolation of skeletal muscle cells by growing them at low confluence in proliferation medium. To induce myoblast differentiation, cells were plated at high confluence and changed to differentiation media. Gene Ontology (GO) analysis of the genes upregulated in HDAC11 KO differentiating myoblasts revealed a statistically significant enrichment of cell cycle-related processes while the downregulated genes in HDAC11 KO differentiating myoblasts were enriched in “muscle system process” and “muscle contraction” GO categories.