Unknown,Transcriptomics,Genomics,Proteomics

Dataset Information

Gene expression profiling of bovine muscle samples upon experimental treatment with dexamethasone and dexamethasone/Clenbuterol combination

ABSTRACT: In the present study, Bovine transcriptome was characterized in order to identify a set of candidate genes potentially useful for rapid detection of illegal use of some growth promoters (GPs) as dexamethasone (DEX) alone or in combination with the B2-agonist clenbuterol (CLEN). A bovine oligo microarray platform (GPL7053) was used to profile gene expression from twelve (12) bovine biceps brachii samples. Isolated and purified total RNAs were individually hybridized to the Agilent bovine V1 4x44k DNA microarray. The comparison of untreated and treated bovine transcriptomes revealed a set of differentially expressed genes. After functional analysis and qPCR validation, 16 genes were found to be differentially expressed among groups (15 downregulated genes & 1 upregulated gene between CTR and DEX and CTR and DEX CLEN groups, respectively). A total of 24 animals was used in this study. Animals were randomly divided into three groups of 8 animals each. The first group was used as a control (CTRL), the second was treated with dexamethasone (DEX) administered via feed 0.75 mg per capita for 42 days (group DEX), and the third one was treated with an increasing dose of Clenbuterol (CLEN) via feed 2 mg per capita during the first week, 4 mg per capita during the second week, and 6 mg per capita during the third and the fourth weeks (28 days in total), in combination with DXM 0.66 mg per capita for 21 days (group DEX+ CLEN).Twelve samples (4/group) out of the 24 were chosen for the microarray analysis based on the highest isolated RNA quality.

ORGANISM(S): Bos taurus

SUBMITTER: Ramy Elgendy

PROVIDER: E-GEOD-61934 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

Transcriptomic analysis of skeletal muscle from beef cattle exposed to illicit schedules containing dexamethasone: identification of new candidate biomarkers and their validation using samples from a field monitoring trial.

Elgendy Ramy R Giantin Mery M Montesissa Clara C Dacasto Mauro M

Food additives & contaminants. Part A, Chemistry, analysis, control, exposure & risk assessment 20150806 9

Growth promoters (GPs) such as the glucocorticoid dexamethasone (DEX) and the β-adrenergic agonist clenbuterol (CLEN) are still used abusively in beef cattle production. Transcriptomic markers for indirect detection of such GPs have been discussed in either experimentally treated animals or commercial samples separately. In the present study we examine the transcriptomic signature of DEX alone or in combination with CLEN in skeletal muscle of experimentally treated beef cattle, and, furthermore, ...[more]

PMID: 26161592

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Project description:In the present study, a oligonucleotide microarray platform is used to compare expression profiles of beef cattle muscle in animals treated with either Dexamethazone (Dex) or Dexamethazone plus 17?-estradiol (Estr) administered at sub-therapeutic dosage, against untreated controls. Seventeen male beef cattle 15-18 months old, around 450 kg mean body weight were randomly allocated in three groups: 6 were untreated (group Ctrl), 5 were administered with dexamethazone via feed 0.75mg/head for 43 days (group D); the last 6 animals were administered via feed for 43 days with Dex (0.75mg/head) and intramuscularly (i.m.) for three times with 17?-oestradiol, 20mg/head, (group DE). Three additional control animals, matching in sex and age, collected in a previous experiment were included in the Ctrl group to increase sample size and to control for biological variation. For each sample, total RNA was extracted from a specific anterior limb muscle (biceps branchii). Data analysis demonstrates that the expression profiles were strongly affected by Dex treatment with hundreds of genes up-regulated with relevant fold-change, whereas the myostatin gene was significantly down-regulated. On the contrary, the administration of Dex-Estr reveals a weaker effect on gene expression. In this study, we analyzed the gene expression profiles of 20 anterior limb muscle samples, 9 collected from untreated controls, 5 collected from cattle treated with Dex and 6 from cattle treated with Dex+Estr using Agilent-015354 Bovine oligo microarray platform (20 arrays, no replicate) based on single-colour detection (Cyanine-3 only). Microarrays are scanned with Agilent scanner G2565BA (barcode on the left, DNA on the back surface, scanned through the glass) at a resolution of 5 microns; all slides are scanned twice at two different sensitivity settings (XDRHi 100% and XDRLo 10%); the scanner software creates a unique ID for each pair of XDR scans and saves it to both scan image files. Feature Extraction 9.5 uses XDR ID to link the pairs of scans together automatically when extracting data. The signal left after all the FE processing steps have been completed is ProcessedSignal that contains the Multiplicatively Detrended, Background-Subtracted Signal

Project description:Purpose: We applied the next-generation sequencing to explore the differential gene expression in CAD cells differentiated with dexamethasone (Dex), compared to the undifferentiated cells using Illumna -based RNA-sequencing. Methods: Total RNAs were extracted from undifferentiated CAD cells and Dex-differentiated cells (5 days). Three independent samples were collected from both experimental groups. Samples then underwent to the sequencing step performed on NextSeq with a 75 base-pair end read length. Quality filtered reads were mapped to the mouse reference genome (GRCm38.p6). Feature Counts software was implemented to obtain raw counts for all mouse genes. The gene counts were used for differential expression analysis with the DESeq2 package to identify differentially expressed genes (DEGs) in the Dex-differentiated CAD cells, as compared to the undifferentiated CAD cells. Results: The sequence reads (expression level) per sample was obtained in a range of 9.8-11.2 million. The high quality of sequence reads was mapped to the mouse reference transcriptome (GRCm38.p6). The gene counts were used for differential expression analysis. The sample-to-sample distance demonstrated that the clustering of the undifferentiated groups was completely separated from the clustering of Dex-differentiated groups. A PCA plot showed that there was a huge variation (PC 1 =99%) between the Dex-differentiated group and the undifferentiated group but less variation was observed within the same group (PC2 = 0%). These statistic profiles indicate that the transcriptomic profile between Dex-differentiated CAD cells and undifferentiated CAD cells had the different pattern. A total of 1,183 differentially expressed genes (DEGs) were identified in the Dex-differentiated CAD cells compared to the undifferentiated CAD cells (false discovery rate < 0.01 and |log2 [fold change] | > 2). Of these, 803 and 380 genes were significantly upregulated and downregulated in the Dex-differentiated group, respectively. Conclusion: Dexamethasone causes extensive changes in gene expression of CAD cells.

Project description:Introduction: Renal ischemia-reperfusion (IR) causes acute kidney injury (AKI) with high mortality and morbidity. The objective of this study was to ameliorate kidney IR injury and identify novel biomarkers for kidney injury and repair. Methods: Left renal ischemia was induced in rats by clamping renal artery for 45 minutes, followed by reperfusion and right nephrectomy. Thirty minutes prior to ischemia, rats (n=8/group) received Valproic Acid (150 mg/kg; VPA), Dexamethasone (3 mg/kg; Dex) or Vehicle (Saline) intraperitoneally. Animals were sacrificed at 3h, 24h or 120h post- IR and blood, urine and kidney were collected. Results: Serum creatinine (mg/dL) at 24 h IR in VPA (2.7±1.8) and Dex (2.3±1.2) was reduced (P<0.05) compared to Vehicle (3.8±0.5). At 3h post-IR, urine albumin (mg/ml) was higher in Vehicle (1.47±0.10), VPA (0.84±0.62) and Dex (1.04±0.73) compared to uninjured/untreated control (0.14±0.26) group. At 24h post-IR urine Lipocalin-2 (µg/ml) was significantly higher (P<0.05) in VPA, Dex and Vehicle groups (9.61-11.36) compared to uninjured/untreated control (0.67±o.29); also, Kidney Injury Molecule-1 (KIM-1; ng/ml) was significantly higher in VPA, Dex and Vehicle groups (13.7-18.7) compared uninjured/untreated control (1.7±1.9). KIM-1 levels were significantly (P<0.05) higher in all groups compared to uninjured/untreated control levels. Histopathology at 3h post IR demonstrated (P<0.05) reduction in ischemic injury in the renal cortex in VPA (Grade 1.6± 1.5) compared to Vehicle (Grade 2.9±1.1) group. Inflammatory cytokines IL1β and IL6 were down-regulated in VPA and Dex groups. BCL2 was higher in VPA group. DNA microarray analysis demonstrated reduced stress response and injury, and improved recovery related gene expression in the kidneys of VPA treated animals. Conclusions: VPA administration reduced kidney IR injury and improved regeneration. KIM-1 and Lipocalin-2 appear to be promising early urine biomarkers of acute ischemic kidney injury. We had three experimental groups. Group A, VPA treatment; Group B, Dexamethasone treatment; and Group C, No treatment (vehicle saline control). Treatments were administered prior to the induction of left renal ischemia. Animals underwent 45 minutes of left renal ischemia, followed by reperfusion, and right nephrectomy as described above. Following reperfusion, animals were sacrificed at 3, 24 or 120 hours (n=8/group). In the 3 hour group, rats were maintained under anesthesia after surgery until sacrifice. In the 24 and 120 hour groups, the rats were recovered and returned to the cages for normal housing. Analgesic buprenorphine (0.05mg/kg) was administered every 12 h for three days post-operatively. After animal sacrifice, urine, blood, and kidney were collected for kidney functional biomarker assays, histology and / or molecular analyses. Urine was obtained via cystocentesis and blood was obtained via the left renal vein. Tissue and urine samples collected from normal (naïve) animals (n=5) were used for baseline measurements. A subset of 46 animals (n = 4-5 per group) were selected for microarray analysis.

Dataset Information

Gene expression profiling of bovine muscle samples upon experimental treatment with dexamethasone and dexamethasone/Clenbuterol combination

Publications

Transcriptomic analysis of skeletal muscle from beef cattle exposed to illicit schedules containing dexamethasone: identification of new candidate biomarkers and their validation using samples from a field monitoring trial.

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