Gene profiling of the early healing rat medial collateral ligament
Ontology highlight
ABSTRACT: The objective of the study was to better understand the mechanism behind scar formation by identifying ECM factors and other unique genes differentially expressed during rat ligament healing via microarray. Rat medial collateral ligaments (MCL) were surgically transected or left intact. MCLs were collected at day 3 or 7 post-injury and used for microarray analysis. Results were compared to the normal intact ligaments. A total of 27 rats were used in the study. Animals were randomly placed in 1 of 3 groups (day 3, day 7, intact CX) with nine animals included with each time (9 rats/group). At the time of collection, MCLs from each time were placed into 3 pools (3 MCLs/pool), resulting in 3 reps per time, and usd for microarray analysis. A total of 9 gene chips were used for the experiment (3 chips/day).
Project description:Rgg-dependent transcriptional regulation in Streptococcus pyogenes strains MGAS5005 and CS101 was analyzed during post-exponential phase of growth Keywords: strain comparion, post-exponential growth, rgg mutant Microarray analysis was performed using RNA samples isolated from wild-type MGAS5005 and CS101 strains as well as their rgg mutant strains during post-exponential phase of growth
Project description:Rgg-dependent transcriptional regulation in SF370 Streptococcus pyogenes strain was analyzed during post-exponential phase of growth Keywords: rgg mutant Microarray analysis was performed using RNA samples isolated from both wild-type SF370 and SF370 rgg mutant strains during post-exponential phase of growth
Project description:The goal was to screen for the expressed genes in Semi-Circular Canal Duct (SCCD) that are related to ion transport and its regulation. The objective was to discover which genes changed expression levels in response to glucocorticoids. Keywords: drug response Primary cultures were incubated for 24 hours in the presence or absence of 100 nM dexamethasone. Four independent cultures from each group were processed for total RNA and submitted for gene array analysis. Although two Samples out of four in each group were done at different time points, we found that there was not much variation between the datasets (by cluster analysis).
Project description:Goal of experiment: Identification of differentially expressed immune genes from male and female BWF1 lupus-prone mice. (Female incidence is higher than male--attempting to find sex hormone regulated genes that may contribute to this difference).
Project description:Staphylococcus aureus is a major human pathogen and resistant to numerous clinically used antibiotics. The first antibiotic developed for S. aureus infections was the nonribosomal petide secondary metabolite penicillin. We discovered cryptic nonribosomal peptide secondary metabolites, the aureusimines, made by S. aureus itself that are not antibiotics, but function as small molecule regulators of virulence factor expression. Using established rules and codes for nonribosomal peptide assembly we predicted these nonribosomal peptides, and used these predictions to identify them from S. aureus culture broths. Functional studies using global microarray and mouse bacteremia models established that the aureusimines control virulence factor expression and are necessary for productive infections. This is the first report of the aureusimines and has important implications for the treatment of drug resistant S. aureus. Targeting aureusimine synthesis may provide novel anti-infectives. Commerically available S. aureus GeneChips (Affymetrix) were used to compare biological replicates of early and late exponential phase wild type (Newman) and aureusimine defective (ausA) organisms.
Project description:Haemophilus parasuis(HPS) is a prominent swine pathogen that causes GlM-CM-$sser's disease characterized by fibrinous polyserositis, meningitis and arthritis; however, the molecular mechanisms underlying disease pathogenesis remains poorly understood, particularly the host counteraction to HPS invasion by the immune system. Here, we investigated the global expression changes in spleen following HPS infection using the Affymetrix Porcine Genechip.Our findings indicate previously unrecognized gene transcription changes in case of HPS infection in vivo and many presumed cascades in the study are clearly merit further investigation. Our data should provide new clues for immune response in mammals and identification of candidate genes related to HPS resistance. All animal tissue collection procedures were performed according to protocols approved by The Hubei Province, PR China for Biological Studies Animal Care and Use Committee. Piglets at 30 days old were determined to be HPS-free by serum indirect haemagglutination test before artificial bacterial challenges. Each piglet was intratracheally challenged with 5M-CM-^W108 colony-forming units (CFU) per millilitre of HPS strain 0165 (serotype 5) . A total of 5 piglets were challenged, and 5 pigs were used as controls. These two groups were raised in isolated facilities. Six microarrays were used in the experiment, corresponding to the RNAs from spleen tissues of three most severe piglets with fibrinous polyserositis, meningitis and arthritis after HPS infection and three controls.
Project description:To test the efficacy of TNFR-Fc and anti-TWEAK mAb treatment alone and in combination Tumor necrosis factor (TNF)-alpha is a major effector in various inflammatory conditions. TNF-like weak inducer of apoptosis (TWEAK) is a member of the TNF superfamily that promotes inflammatory tissue damage through its receptor, FGF-inducible molecule 14 (Fn14). Since both TWEAK and TNF-alpha have been shown to mediate pathological responses through inter-dependent or independent pathways by in vitro, the potential interplay of these pathways was investigated in a mouse colitis model. Acute colitis was induced by rectal injection of trinitrobenzene sulfonic acid (TNBS), with administration of control IgG, TNF receptor (TNFR)-Ig chimeric protein, anti-TWEAK monoclonal antibody, or the combination of TNFR-Ig and anti-TWEAK antibody. On day 4, disease severity was evaluated and gene expression profiling was analyzed using whole colon tissue. Levels of transcript of TWEAK, Fn14 and NF-kB-related molecules were measured in purified colon epithelial cells (ECs). NF-kB activation was investigated with Western blot and immunohistochemical analysis. As a result, activation of the canonical, but not noncanonical NF-kB pathway was the hallmark of inflammatory responses in this model. Inflammation induced upregulation of Fn14 only in ECs but not in other cell types. Combination treatment of TNFR-Ig and anti-TWEAK antibody synergistically reduced disease severity in comparison with the control antibody or single agent treatment. Gene expression profile of the colon indicated downregulation of canonical NF-kB pathway with combination treatment. In conclusion, synergistic activation of canonical NF-kB by TWEAK and TNF-alpha is critical for the induction of inflammatory tissue damage in acute inflammation. TNBS colitis was induced by intrarectal administration of a 2 % solution of TNBS in phosphate-buffered saline (PBS): ethanol (1:1). For acute inflammatory responses, 70 M-NM-<g/g body weight of TNBS was given on day 0 and animals sacrificed on day 4. One hour prior to administration of TNBS, groups of mice were injected i.p. with the control IgG2a mAb (anti-human CD20) (10 mg/kg), TNFR-Ig (0.3 mg/kg), anti-TWEAK (mP2D10, 10 mg/kg), the combination of TNFR-Fc (0.3 mg/kg) and anti-TWEAK mP2D10 (10 mg/kg), or were untreated. For single agent and combination treatments groups, 0.3 mg/kg TNFR-Ig was employed, since 1 mg/kg of TNFR-Ig markedly ameliorated TNBS colitis but 0.3 mg/kg was much less effective as monitored by the effect on colon length and body weight. Colon tissue was rolled and snap-frozen in liquid nitrogen. Specimens for RNA extraction were cut from the frozen rolled colon. A set of experiments using 5-8 mice for each experimental group was performed twice, labelled 7 and 10 in the CEL files. Total number of mice for each experimental condition was as follows; untreated TNBS colitis group, 8; control antibody, 15; TNFR-Ig, 12; anti-TWEAK mAb, 14; combination of TNFR-Ig and anti-TWEAK mAb, 15.
Project description:Gene expression profiling in soybean under aluminum stress: Transcriptome response to Al stress in roots of Al-tolerant genotype (PI 416937). Aluminum (Al) toxicity is a major constraint of crop production on acid soils. Many commercial soybean cultivars and advanced breeding lines have been evaluated for Al tolerance. Aluminum tolerance is quantitatively inherited trait in soybean making it difficult for genetic improvement. Understanding the molecular and genetic mechanisms of tolerance is crucial for developing efficient and effective programs aimed at improving Al tolerance trait The molecular mechanisms of Al tolerance is poorly understood in soybean. The objective of the research was to identify candidate aluminum tolerance genes in soybean Al-tolerant soybean genotype PI 416937 seedlings were exposed to zero or 10 µM Al in growth chamber under hydroponic conditions for four time span of 2, 12, 48 and 72 hrs in a randomized complete block design with three replications. Microarray analysis was made on mRNA isolated from 1 cm log tap root tips using Affymetrix soybean array with over 68,000 probe sets Glycine max L and wild soybean combined. Both novel and known genes were discovered in response to Al treatment. They include Al tolerance relevant proteins, families of transcription factors, zinc finger, bZIP, WRKY, MYB, ADR6, and NAC domain proteins were induced likely regulating Al tolerance downstream genes. Stress related proteins, cytochrome P450, glutathione-s transferase, glutaredoxin family and ascorbic acid biosynthesis protein were induced as signatures of cellular detoxification mechanisms. An ABC type multidrug resistance protein that could act as citrate transporter or Al exporter was up-regulated, a key Al tolerance mechanisms in several species. A cell wall loosening enzyme endoxylglucan hydrolases were also up-regulated probably reversing the wall rigidification caused by Al and promoting root growth under Al stress. Phytosulfokines growth factor involved in cell division and proliferation was up-regulated likely as a direct counter action to Al toxicity which inhibits root growth by limiting cell division and elongation. In conclusion, the Al tolerance candidate genes identified herein are potential targets for future genetic engineering and molecular breeding work on Al tolerance trait in soybean which in turn would contribute to gain in soybean productivity on acid soils. One genotype PI 416937 (p); four time points: (2, 12, 48, and 72 hrs), with replicates (2 or 3)
Project description:By carrying out a systematic structure/function study of the RANK cytoplasmic domain, we previously identified a specific 4-a.a. RANK motif (IVVY535-538) which plays a critical role in osteoclastogenesis by mediating commitment of macrophages to the osteoclast lineage. We have recently validated the role of this IVVY motif in osteoclastogenesis in vivo by generating knockin (KI) mice bearing inactivating mutations in the RANK IVVY motif. This microarray experiment was performed to determine whether the IVVY motif is involved in regulating gene expression in osteoclastogenesis. We used microarrays to detail the global programme of gene expression underlying cellularisation and identified distinct classes of up-regulated genes during this process. Bone marrow macrophages isolated from wild-type (WT) or knockin (KI) mice were plated in 60-mm tissue culture dishes and treated with M-CSF (44ng/ml) and RANKL (100ng/ml) for 24 hours. Each genotype has three triplicates. Total RNA was isolated for microarray analysis using mouse chips (type 430.2.0) at the Microarray Shared Facility at the University of Alabama at Birmingham.
Project description:Gene expression profiling in soybean under aluminum stress: genes differentially expressed between Al-tolerant and Al-sensitive genotypes. Aluminum toxicity is the most important constraint of crop production on acid soils. Understanding the molecular and genetic mechanisms of tolerance is crucial for developing efficient breeding programs to improve Al tolerance. This research was undertaken to identify candidate Al-tolerance genes in soybean. Two soybean genotypes PI 416937 (Al-tolerant) and Young (Al-sensitive) seedlings were exposed to zero or 10 µM Al in a growth chamber under hydroponic conditions for four time spans of 2, 12, 48 or 72 hrs. Microarray analysis was made on mRNA isolated from 1 cm long tap root tips using an Affymetrix soybean genome array. Both novel and previously reported aluminum-responsive genes were identified. The differentially expressed genes were enriched for metabolism, stress response and transporters. Multiple putative Al-tolerance genes uniquely induced in the tolerant genotype includes the up-regulation of previously identified transcription factors auxin down regulated-like protein (ADR6-like) and basic leucine zipper (bZIP 94), sulfur transmembrane transport protein and lipid transfer protein (Sec 14 ) and novel genes that include rare cold inducible protein (RCI2B ), GPI-transamidase, malonyl-COA: Isoflavone 7-O-glucoside-6˝-O-malontransferase, a cell proliferation protein (WPP2), Oleosin protein, pectinestrease inhibitor, and impaired sucrose induction1. The genes identified in this study will be utilized as important genetic resources for future improvement of Al tolerance in soybean. Key words: Soybean, Al tolerance, gene expression, microarray Two genotypes: PI 416937 (p) and Young (y); two treatments: aluminum or untreated; four time points: 2, 12, 48, and 72 hrs; 2 or 3 replicates.