Project description:Using NGS, we report the impact of cardiovascular risk factors (CRF) including hypercholesteolemia, aging and smoking compare to healthy control mice, on the differential-expression profile of miRNAs in proangiogenic cells (PACs) isolated during ischemia-induced neovascularization in a mouse model of hindlimb ischemia. Data of total miRNA expression are presented as normalized copy number: read per millions of read (RPM). Hindlimb ischemia was surgicaly induced by feromal artery ablation and the animal was sacrified 2 day after ischemic surgery for PACs isolation. After 4 days of culture, PACs RNAs were extracted to perform NGS.
Project description:We identified the genes regulated by hindlimb ischemia in leukocytes and hematopoietic stem and progenitor cells isolated from the bone marrow in mice. We selected some of these genes based on their expression and further evaluated their functions using a wise array of techniques.
Project description:Using NGS, we report the differential-expression profiling of miRNAs in hypercholesterolemic/atherosclerotic conditions in hindlinb ischemic muscle compare to healthy control mice. Data of total miRNA expression are presented as normalized copy number: read per millions of read (RPM). Hindlimb ischemia was surgicaly induced by feromal artery ablation and the animal was sacrified 1 day after surgery for RNA collection.
Project description:Purpose: The aim of this study is to compare the plasma miRNA profile between mice subject to myocardial ischemia and reperfusion and mice subject to sham operation. Methods: 8 to 10-week old C57BL/6 mice underwent myocardial ischemia and reperfusion (MIR) or Sham operation. Plasma RNA was isolated using Trizol LS reagent 4h post-surgery. NGS cDNA libraries were prepared using Norgen Biotek Small RNA Library Prep Kit. Library quality was validated prior to sequencing on an Illumina NextSeq 500 platform.
Project description:Transient brain ischemia massively activates global SUMOylation. A large fraction of SUMO targets are nuclear proteins involved in gene expression. However, gene expression profile modulated by ischemia-induced SUMOylation has not been studied. Using a SUMO knockdown (SUMO-KD) mouse model and microarray technique, we investigated how SUMOylation modulated gene expression after brain ischemia. Wild-type and SUMO-KD mice were subjected to transient forebrain ischemia or sham surgery. The hippocampal CA1 subfield samples collected at 3 hours reperfusion were analyzed using Affymetrix microarrays.
Project description:Ischemia/reperfusion injuries is a known complication to hepatic surgery. Ischemic pre- (IPC) and postconditioning (IPO) protects the liver against ischemia/reperfusion-injuries. Expression profiling were performed on liver biopsies seeking to identify molecular mediators of the protective properties. 48 rats were divided into 5 groups; sham (n=8), IRI (n=10), IPC (n=10), IPO (n=10) and IPC+IPO (n=10). All rats except sham rats were subjected to 30 min of total liver ischemia and 30 min of reperfusion before liver biopsies were sampled. In the IPC group, liver ischemia was preceded by 10 min of hepatic ischemia, followed by 10 min of reperfusion. IPO were performed by three cycles of 30 sec of reperfusion and 30 sec of ischemia, applied immediately after the 30 min of total liver ischemia. In the IPC+IPO group the two interventions were combined.
Project description:We aimed to develop a novel chronic and severe hindlimb ischemia mice model to properly evaluate the therapeutic effects of drug candidates in translational research for critical limb ischemia treatments. We used microarray to compare the gene expression patterns of gastrocnemius muscles at 1st week and 9th week after hindlimb ischemia model operation.
Project description:Experimental Design: 1. The goal of the experiment: Age-Related Genome Expression Profiles During Hepatic Ischemia 2. Brief description of the experiment: Hepatic ischemia/reperfusion (I/R) injury is a complication of liver surgery, transplantation and shock and differs with age. In the present study, we sought to determine if the age-dependent response to I/R injury was related to differential gene expression during the ischemic period. Total RNA of young (4-5 weeks) and adult (12-14 months) mice undergoing sham surgery or partial hepatic ischemia for 30, 60, or 90 minutes was analyzed by Affymetrix microarray. Gene expression was filtered based on a change in expression of 1.5-fold relative to respective controls and then analyzed by ANOVA. Significant differences in gene expression were observed between age groups. In young mice, 169 genes were down-regulated, whereas adult mice had 1167 genes down-regulated. Of these, only 28 genes were down-regulated in both young and adult mice. Far fewer genes had increased expression during ischemia. Sixty genes in young mice and 51 genes in adult mice were up-regulated. Of those, none were up-regulated in both young and adult mice. There were no distinct functional patterns of gene regulation in either age group. The data demonstrate significant and widespread changes in hepatic gene expression during the ischemic period that may be important to the age-dependent response to I/R injury. Keywords: treated vs non treated 2 age groups of 12 young and 12 adult mice that underwent either 30, 60 or 90 minutes of partial ischemia (n=3) or sham operation (n=3) We used microarray to uncover the genomic response during different time points of ischemia
Project description:Peripheral Artery Disease is caused by the restriction or occlusion of arteries supplying the leg. Better understanding of the molecular mechanisms underpinning tissue response to acute and chronic ischemia is urgently needed to improve therapeutic options. The aim of this study is understanding miR-210 regulation and role in a mouse model of hindlimb ischemia. To investigate miR-210 function, mice were injected with a miR-210 complementary LNA-oligonucleotide (anti-miR-210). Then, the left femoral artery was dissected in order to induce unilateral hindlimb ischemia. Mice were sacrified 3 days later and gene expression profiles of gastrocnemius muscles were obatained.
Project description:Depending on the severity of a CNS injury, in humans as well as in experimental animal models certain degrees of functional recovery are observed. Rodent studies have provided much evidence for underlying anatomical changes involving neurite outgrowth of both spared and axotomized fiber tracts, also referred to as sprouting. Following a thoracic spinal cord injury (SCI) in adult rats, axotomized fibers of the hindlimb corticospinal tract (CST) were found to sprout into the cervical spinal cord at different rostrocaudal levels. Anterograde tracings showed that these new collaterals of motor cortical hindlimb fibers project into the intermediate laminae of the cervical grey matter, particularly pronounced at the level of cervical segment three (C3). The molecular cues which induce the growth and guide these sprouts of hindlimb CST fibers are unknown. We hypothesize that sectreted factors and guidance and adhesion molecules are specifically expressed in the intermediate laminae of the cervical spinal cord to direct and target the ingrowth of the sprouting hindlimb CST fibers. To test our hypothesis the transcription profile of the target region of sprouts was investigated with RNA sequencing of total RNA extracted from the intermediate laminae (IV-VII) of the spinal gray matter at cervical level C3. Four major comparisons were performed, which were then again compared to each other. The first comparison was between samples of animals with an SCI sacrificed at four different time points after injury (1, 3, 6, 12 weeks after injury). The second and third comparison was between samples of rats with a SCI compared to their sham controls sacrificed at the same time point after surgery (1 week or 12 weeks respectively). The fourth comparison was between samples from animals of the two control groups, rats with a sham surgery which were either sacrificed at one or at twelve weeks after injury. We found that most changes in gene expression ocurred at one week after SCI and least changes occured at twelve weeks after SCI, suggesting that the largest effects on a global level of gene expression occur during early time points after injury. Interestingly, also we also found changes in gene expression when comparing the samples from the two control groups, which suggests that sham surgery consisting of a laminectomy is sufficient to induce transcriptional changes in the spinal cord. This is the first study to provide insight into transcriptional changes in the target region of sprouts investigating the role of the cervical spinal cord (target region) for sprouting of hindlimb CST fibers.