Project description:To identify atrophy genes directly targeted by Bcl-3 transactivator at a genome wide level, we performed whole transcript expression array and ChIP-seq for muscles from weight bearing or 5-day hind limb unloaded mice. Genes that showed increased expression with unloading and a Bcl-3 peak in the promoter (from ChIP-seq data) were considered as Bcl-3 direct targets during disuse atrophy. Using ChIP array, we identified 241 direct targets for Bcl-3. Our data describe Bcl-3 as a global regulator both of the proteolysis and the change in energy metabolism that are essential components of muscle atrophy due to disuse. Disuse skeletal muscle atrophy was induced by hind limb unloading. Weight bearing (WB) or 5-day hind limb unloaded (HU) muscles were harvested for total RNA isolation and processed for whole transcript expression profiling. We chose to examine gene expression and Bcl-3 binding from 5-day unloaded muscles because our previous time course study of disuse atrophy suggested that most genes are differentially regulated at this time point, and thus, would best represent the time for Bcl-3 binding to the gene targets of the NF-kB transcriptional network.
Project description:Long non-coding RNAs (lncRNAs) play important roles in diverse biological processes. However, the landscape of lncRNAs is largely unclear in Sus scrofa. Here we performed stranded RNA-seq on total RNA libraries from over 100 samples of Sus scrofa tissues. We identified 10,813 lncRNAs in Sus scrofa, of which 9,075 are novel. 57% of these lncRNAs were conserved in both human and mouse. These conserved lncRNAs tend to be more tissue-specific than pig-specific lncRNAs, and enriched in reproducible organs (i.e. testis and ovary). We characterized a group of lncRNAs potentially involved in the skeletal muscle development. One such lncRNA, a homolog of maternally expressed gene 3 (MEG3), was specifically expressed in the skeletal muscle at early developmental stage. And its expression pattern is conserved in pig and mouse. By over-expressing and knocking down MEG3 in mouse myoblast cell lines, we demonstrated its novel function as a myoblast proliferation suppressor.
Project description:We took advantage of a dystrophic mouse model of transient macrophage-depletion, mdxITGAM-DTR mice, in order to analyze the role of macrophage in skeletal muscle regeneration. We generated the transcriptome of satellite cells (SCs) and alpha7Sca1 cells purified by cell sorting from mdxITGAM-DTR mice. The mice were treated, by intramuscular injection, with PBS, as vehicle, or with Diphtheria toxin (DT) in order to achieve the macrophage depletion form hind-limb muscle We described a shift in identity of muscle stem cells dependent on the crosstalk between macrophages and satellite cells. Indeed macrophage depletion determines an exacerbated dystrophic phenotype associated with adipogenic conversion of SCs and reduction of the SC pool.
Project description:The aim of the study was to determine the protein composition of cornified claws of the western clawed frog (Xenopus tropicalis) in comparison to clawless toe tips and back skin. Cornified claws develop on toes I, II, III of the hind limbs, which we refer to as hind limb inner (HI) toes. Toes IV, V of the hind limbs, here referred to as hind limb outer (HO) toes lack claws. Proteins were prepared from HI toe tips including claws, HO toe tips and back skin (BSK) of frogs each (F1, F2, F3) and subjected to proteomic analysis.
Project description:Comparison of NRA derived from SCA1+, CD45- MACS purified putative muscle derived stem cells vs. RNA derived from preplating of hind limb muscle cells. Keywords: other
Project description:The goal of this study was to identify changes in muscle gene expression that may contribute to loss of adaptability of old muscle. Muscle atrophy was induced in young adult (6-month) and old (32-month) male Brown Norway/F344 rats by two weeks of hind limb suspension (HS) and soleus muscles were analyzed by cDNA microarrays. We conclude that a cold shock response may be part of a compensatory mechanism in muscles undergoing atrophy to preserve remaining muscle mass and that RBM3 may be a therapeutic target to prevent muscle loss.
Project description:Global gene expression patterns were determined from microarray results on day 1, 3, 5, 7, 10 and 14 during plantaris muscle regrowth following two weeks of hind limb suspension in young adult mice (5 months). For each time point, one Affymetrix chip (Mouse Gene 1.0 ST) was used with 100 ng of total RNA derived from a pooled sample of the left plantaris muscle from six animals.
Project description:The current study aimed to address the hypothesis that programmed expression of key miRNAs in skeletal muscle mediates the development of insulin resistance, and consequently long-term health. We thus examined microRNA signatures in skeletal muscle of programmed insulin resistant rats offspring from high fat-fed dams vs control offspring from chow fed dams. Skeletal muscle (soleus) was collected from the hind limb of 1 year old male offspring (6 from control dams, 6 from high fat-fed dams) . Ramaciotti Centre for Genomics (UNSW, sydney, Australia)