Combination of high-dimensional approaches for skeletal muscle tissue cartography
Ontology highlight
ABSTRACT:
ORGANISM(S): Mus musculus
PROVIDER: GSE110878 | GEO | 2019/03/30
REPOSITORIES: GEO
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Project description:Satellite cells are responsible for the long-term regenerative capacity of adult skeletal muscle. The diminished muscle performance and regenerative capacity of aged muscle is thought to reflect progressive fibrosis and atrophy. Whether this reduction in muscle competency also involves a diminishment in the intrinsic regulation of satellite cell self-renewal remains unknown. We used microarray to identify gene expression changes underlying the marked reduction in the capacity of satellite cells to self-renew, contribute to regeneration and repopulate the niche as they age. Skeletal muscles from heterozygous Pax7-ZsGreen mice were isolated at defined stages: E17.5 (fetal - whole forelimb and hindlimb), postnatal day 21 (adolescent - hindlimb), 2-3 month old (young adult - hindlimb) and >1 year old (older adult - hindlimb) mice. ZsGreen-positive skeletal muscle satellite cells were isolated by FACS and pooled (fetal n=4, adolescent n=6, young adult n=8 and older adult n=8 mice).
2014-05-26 | E-GEOD-47401 | biostudies-arrayexpress
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2016-07-21 | E-GEOD-83555 | biostudies-arrayexpress
Project description:Skeletal muscle tissues are comprised of muscle fibers, muscle stem cells (MuSCs), immune cells, endothelial cells, motor/sensory nerves with Schwann cells, and interstitial mesenchymal cells. Each cell population plays a unique role in muscle physiology and homeostasis. The mesenchymal progenitors in the skeletal muscle have been identified as a subpopulation that reside between muscle fibers, whose role is to coordinate muscle regeneration in a finely tuned manner by supporting the expansion and differentiation of MuSCs. These cells have been referred to as fibro/adipogenic progenitor cells (FAPs), termed according to their differentiation potentials in specific in vitro culture conditions and known to cause fibrous tissue and fatty degeneration under pathological environments and result in compromised muscle function. To gain insight into how Bap1 plays a role in skeletal muscle homeostasis, we performed RNA sequencing analysis using freshly isolated Sca1 positive cells from 1.5-week-old WT or cKO hindlimb muscles
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Project description:Skeletal muscle is a post-mitotic tissue that exhibits an extremely low turnover in the absence of disease or injury. At the same time, muscle possesses remarkable regenerative capacity mediated by satellite cells (SCs) that reside in close association with individual myofibers, underneath the fiber’s basal lamina. Consistent with the low turnover of the muscle, SCs in adult animals are mitotically quiescent and therefore provide an excellent model to study stem cell quiescence. As an organism grows older, the resident stem cells are exposed to a deteriorating environment and experience chronological aging. In stem cells with high turnover, the effects of chronological aging are superimposed upon the effects of the replicative aging that results from DNA replication and cell division. On the contrary, SCs experience minimal replicative aging due to their low turnover. They are thus a good model to study the consequence of chronological aging of quiescent stem cells. We performed microarray analysis of quiescent and activated SCs from both young and aged mice to understand the global gene expression profile underlying stem cell properties such as quiecence and self-renewal, and to understand how the transcriptome of a quiescent stem cell pouplation changes with age. VCAM+/CD31-/CD45-/Sca1- quiescent satellite cells (QSCs) were isolated by FACS from hindlimb muscle of uninjured 2-3- or 22-24-month old mice. Activated satellite cells (ASCs) were isolated from hindlimb muscles of BaCl2-injured mice of the same age 36, 60 and 84 hours after injury using the same cell surface marker combination. YFP-expressing cells were isolated from 2-3-month old Pax7CreER/+; ROSA26eYFP/+ mice in which satellite cells are labeled geneticall by YFP expression. Total RNA was extracted from cells with the Trizol reagent according to manufacturer's instructions. RNA was then processed and assayed with Affymetrix Mouse Gene 1.0 ST arrays.
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Project description:Following skeletal muscle injury, muscle stem cells (satellite cells) are activated, proliferate, and differentiate to form myofibers. We show that mRNA decay protein AUF1 regulates satellite cell function through targeted degradation of specific mRNAs. AUF1 targets certain mRNAs containing 3 AU-rich elements (AREs) for rapid decay. Auf1-/- (KO) mice undergo accelerated skeletal muscle wasting with age and impaired muscle repair following injury. Satellite cell mRNA analysis and regeneration studies demonstrate that auf1-/- satellite cell self-renewal is impaired due to increased stability and overexpression of ARE-mRNAs. Control of ARE-mRNA decay by AUF1 and potentially other ARE-binding proteins represents a mechanism for adult stem cell regulation and is implicated in human muscle wasting diseases. We report the RNA transcript expression profiles from sorted satellite cells isolated from wild type (WT) and AUF1-null (KO) mice hindlimb muscles
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Project description:Experimental cell therapies for skeletal muscle conditions have shown little success, primarily because they used committed myogenic progenitors rather than true muscle stem cells, known as satellite cells. Here we present a method to generate in vitro–derived satellite cells (idSCs) from skeletal muscle tissue. When transplanted in small numbers into mouse muscle, mouse idSCs fuse into myofibers, repopulate the satellite cell niche, self-renew, support multiple rounds of muscle regeneration, and improve force production on par with freshly isolated satellite cells. in damaged skeletal muscle. We profiled the epigenomic and transcriptional differences between idSCs and satellite cells, and used these differences to identify core signaling pathways and genes that confer idSC functionality. Finally, fromhuman muscle biopsies we successfully generated satellite cell–like cells in vitro. After further development, idSCs may provide a scalable source of cells for the treatment of genetic muscle disorders, trauma-induced muscle damage and age-related muscle weakness.
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Project description:High regenerative capacity of adult skeletal muscle relies on a self-renewing depot of adult stem cells, termed muscle satellite cells (MSCs). A novel MSC line was isolated from the rat levator ani muscle and termed Levator Ani Satellite Cells (LASCs). Androgen, a known mediator of overall body composition and specifically skeletal muscle mass, has been shown to regulate MSCs. The use of non-steroidal androgen receptor agonists (NARA) aims to retain the beneficial influence of androgen on skeletal muscle, while circumventing undesirable cardiovascular and prostate-related side-effects. Primary objectives: 1) Identify biomarkers of satellite cell growth and differentiation, 2) Characterize a novel muscle satellite cell line, 3) Understand the effects of androgen (NARA) on rat satellite cell activation and recruitment. LASCs will be treated with one of the following conditions (N=4 for all groups): 1) 0.2% DMSO, 4 hours; 2) 10 nM NARA, 4 hours; 3) 0.2% DMSO, 48 hours; 4) 10nM NARA, 48 hours
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Project description:Purpose: To study Nf1 regulated pathways in controlling juvenile satellite cells quiescence induction. Methods: We sequenced total mRNAs isolated from juvenile satellite cells FACS sorted from postnatal day 7 Control (Nf1flox/+ Myf5Cre+) and KO (Nf1flox/flox Myf5Cre+) mouse hindlimb skeletal muscle. Results: Our study suggests that Nf1 is required for maintaining the balance between quiescence induction and amplification / differentiation in the postnatal MP pool. Conclusions: The tumor suppressor Nf1 maintains the myogenic progenitor (MP) pool during postnatal muscle development.
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