Genome-wide profiling of muscle Treg cells from IL-33 treated mice
ABSTRACT: We report gene expression of Treg cells isolated from injured muscle in IL-33 vs PBS treated mice. Male Foxp3-GFP C57BL/6 reporter (2 months old) mice were injured intramuscularly with cardiotoxin/rIL-33 (0.3 ug/muscle). Tregs were sorted directly into Trizol from injured muscle 4 days post-injury. Gene expression profiling of muscle Tregs from IL-33 vs PBS injured mice.
Project description:We report age-related gene expression of Treg cells isolated from injured muscle and spleen. Male C57BL/6 Foxp3-GFP reporter mice were injured intramuscularly with cardiotoxin. Tregs were sorted directly into Trizol from injured muscle and spleen 4 days post-injury. Gene expression profiling of muscle and splenic Tregs from 2- vs >6-month old mice (biological duplicate for each).
Project description:We compare the effect of IL-33 vs PBS on muscle regneration from 22-month old mice 8 days post-injury. These analyses from whole muscle provide insight into the role of IL-33 on regeneration in the context of aging. The tibialis anterior muscle from male C57BL/6 mice (22 months old) were cryo-injured using a liquid nitrogen chilled metal probe for 8 seconds. Mice received 2 ug of rIL-33 via intraparitoneal injection the day prior and the day following cryoinjury. 8 days post-injury whole muscle was harvested and snap frozen in liquid nitrogen. Whole muscle was then homogenized directly in TriZol and gene expression profiling was performed on Affymetrix Mouse Gene 1.0 ST Arrays.
Project description:Utilizing glycerol and cardiotoxin (CTX) injections in the tibialis anterior muscles of M. musculus provides models of skeletal muscle damages followed by skeletal muscle regeneration. In particular, glycerol-induced muscle regeneration is known to be associated with ectopic adipogenesis. We characterized genome-wide expression profiles of tibialis anterior muscles from wild-type mice injured by either glycerol or CTX injection. Our goal was to detect gene expression changes during the time course of glycerol-induced and CTX-induced muscle regeneration models, that can lead to ectopic adipocyte accumulation. Tibialis anterior muscle of 12 week old wildtype C57BL/6J male mice injected i.m with either glycerol or CTX were collected 3, 7, 14 or 21 days after injection. 6 biological replicates were used for each time points. Please note that a few replicate samples did not pass QC test, thus, were removed from the submission. Unchallenged tibialis anterior muscles from a group of 5 wildtype C57BL/6J mice were also as a control. Total RNA was extracted, QCed and hybridized to Affymetrix microarrays.
Project description:Global gene expression analysis of injured skeletal muscle showed that amphiregulin (Areg), a growth factor over-expressed by muscle Treg cells, enhances muscle regeneration both in the presence and in the absence of Tregs. Foxp3-DTR+ and Foxp3-DTR- mice were injured with cardiotoxin in TA muscle at day 0 and treated with diphtheria toxin every other day from day 0 until dissection. Amphiregulin or PBS were administered im on day 0 and ip every other day until dissection. TA muscle was flash-frozen and RNA was extracted using Trizol. RNA from whole tissue samples was amplified, labeled, and hybridized to Affymetrix Mouse Gene 1.0 ST Arrays.
Project description:Temporal expression profiling was utilized to define transcriptional regulatory pathways in vivo in a mouse muscle regeneration model. Potential downstream targets of MyoD were identified by temporal expression, promoter data base mining, and gel shift assays; Slug and calpain 6 were identified as novel MyoD targets. Slug, a member of the snail/slug family of zinc finger transcriptional repressors critical for mesoderm/ectoderm development, was further shown to be a downstream target by using promoter/reporter constructs and demonstration of defective muscle regeneration in Slug null mice.
Project description:A phenotypically and functionally distinct population of CD4+ Foxp3+ T cells (Tregs) rapidly accumulates in acutely injured skeletal muscle of mice, just as invading myeloid-lineage cells switch from a pro-inflammatory to a pro-regenerative state. Analysis of gene expression of Tregs and CD4+Foxp3- T cells (Tconvs) from injured muscle and spleen revealed that the transcriptome of muscle Treg cells is distinct from that of splenic Tregs. A set of genes is uniquely expressed by muscle Tregs, while another set is over-expressed by the two muscle populations vis-à-vis their two spleen counterparts. 6 wk-old Foxp3-ires-GFP mice were injured in skeletal muscles with cardiotoxin. Four and fourteen days later, Tregs and Tconvs from spleen and muscle were double-sorted into Trizol. To reduce variability, cells from multiple mice were pooled for sorting, and three replicates were generated for all groups. RNA from 1.5-2.5 x 104 cells was amplified, labeled, and hybridized to Affymetrix Mouse Gene 1.0 ST Arrays.
Project description:We have employed genome-wide transcriptome analysis to characterise the changes in expression that occur between resting and regenerating muscle, and the influence p38-alpha has on these expression profiles.
Project description:Skeletal muscle has great regenerative capacity, which is dependent on muscle stem cells, also known as satellite cells. We established the purification system of myogenic cells. We used microarrays to identify specific genes during the muscle regenerative process. Myogenic cells were purified at regenerative stages for RNA extraction and hybridization on Affymetrix microarrays. CTX-2d and CTX-5d are the samples derived from injured muscles 2 days and 5 days after cardiotoxin-injection, respectively.
Project description:The aim of the experiment is to compare the effect of two different calcineurin A isoforms on skeletal muscle in uninjured mice and during skeletal muscle regeneration (after cardiotoxin injection). The transgenic mice express CnAbeta1 or CnAalpha under the control of the myosin light chain promoter and enhancer.
Project description:Muscle injury was elicited by cardiotoxin injection into the tibialis anterior muscle. Macrophages were isolated 2 days post-injury from the regenerating muscle. We used microarray to obtain global gene expression data of muscle-derived tissue macrophage subsets. Tissue macrophages were collected from regenerating muscle samples of three animals, Ly6C+ F4/80low and Ly6C- F4/80high macrophage subsets were sorted. The global gene expression patterns of distinct macrophage subsets were analyzed on Affymetrix microarrays.