Unknown,Transcriptomics,Genomics,Proteomics

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Transcriptome-wide microRNA expression of white blood cells before and after intense cardiorespiratory exercise

ABSTRACT: Regular physical exercise reduces cardiovascular disease (CVD) morbidity and mortality through several biological mechanisms. We were interested in white blood cell microRNA expression in response to exercise. Ten healthy adult males (19-39 years old) undertook 30 minutes of continuous treadmill running at 80% of maximal oxygen uptake (VO2max). Blood samples were taken before and immediately after exercise. Whole-genome microRNAs were then analysed in the extracted RNA. Microarrays (one per sample, with no pooling) were performed on ten healthy adults (19-39 years old) who undertook 30 minutes of continuous treadmill running at 80% of maximal oxygen uptake (VO2max). Blood samples were taken before and immediately after exercise.Whole blood was collected from arm vein, and white blood cells were separated and preserved in liquid nitrogen and later transferred to a M-bM-^@M-^S80C freezer. After extraction of RNA, cRNA was prepared and arrays performed using the Agilent Human miRNA Microarray Kit Release 16.0 performed at the Ramaciotti Gene Function Analysis facility, University of New South Wales in Sydney, Australia.

ORGANISM(S): Homo sapiens

SUBMITTER: Francine Marques

PROVIDER: E-GEOD-45041 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:Subjects Eleven male volunteers (mean age 42.3 + 7.4 years; weight 73.9 + 7.7 kg, height 176.9 + 4.0 cm, body mass index (BMI) 23.6 + 2.1 kg/m2 and body fat: 17.7 + 4.3%) participated in this experiment. They were recruited among experienced ultra-endurance runners and all of them had run at least a race longer than 24 h or 4100 km. On average, they had 15.3 + 7.1 years of training history in running and 7.1 + 4.4 years of ultra-endurance experience. They reported to run an average of 80.5 + 11.7 km/week. Written informed consent was obtained from the subjects. The study was conducted according to the Declaration of Helsinki. The protocol has been approved by the local ethics committee (Comité de Protection des Personnes Sud-Est 1, France) and registered in http:// clinicaltrial.gov (# NCT 00428779). Procedures The subjects visited the laboratory twice, with each session separated by 3–4 weeks. All subjects had run at least three times on a motorized treadmill before taking part in the experiment. In the first session, body mass, height and percentage of body fat (measurements of skin-fold thickness) were measured. The subjects performed a maximal test on a motorized treadmill (Gymrol S2500, HEF Tecmachine, Andrezieux-Boutheon, France), to determine the anaerobic threshold VO2max and the velocity associated with VO2max. The initial velocity was set at 10 km/h and the first stage lasted 6 min. Then, the test consisted of a maximal discontinuous incremental test (slope 50%), where the speed was progressively increased by 1.5 km/h every 3 min, followed by 1 min of rest for the collection of blood samples from the finger tips. The second session consisted of the 24-h treadmill ultra-endurance running protocol (24TR). Subjects were asked to refrain from strenuous exercise for a week before the 24TR. The day of the experiment, all subjects ate the same lunch at noon. About 2 h before starting, a muscle biopsy was taken. Pre-exercise muscle biopsy samples (120 mg) were collected under local anesthesia from the superficial portion of the left vastus lateralis muscle using a percutaneous technique [Henriksson (1979) Acta Neurol Scand 59: 317–323]. A large well-organized fascicle of fibers was oriented and included in an embedding medium (Cryomount; Histolab, Göteborg, Sweden), frozen in isopentane, cooled to its freezing point in liquid nitrogen and stored in liquid nitrogen until further cryostat sectioning. The remaining pieces of the sample were rapidly frozen and stored in liquid nitrogen until protein analyses were performed. After the muscle biopsy procedure, the subjects rested for approximately 2 h and then they were asked to start the 24TR. Ten minutes after the start of the 24TR, subjects were asked to run 4 min at 8 km/h for measurements of running economy (RE). A slow speed was chosen for RE because the average speed over the 24TR was 7.9 + 1.0 km/h, and 8 km/h is representative of the pace over such an extreme exercise [Millet et al. (2009) Scand J Med Sci Sports 21: 54-61]. The food and water intakes during the 24TR were managed by the subjects as in a normal race. Post-exercise muscle biopsy samples (120 mg) were then collected as described above.

Project description:White blood cells (WBCs) express tens of thousands of genes. Their expression levels are modified by genetic and external factors. The purpose of the present study was to investigate the effects of acute exercise on gene expression profiles (GEPs) of WBCs and to identify suitable genes which may serve as surrogate markers for monitoring exercise and training load. Five male probands performed an exhaustive treadmill test (ET) at 80% of their VO2max, and a moderate treadmill test (MT) at 60% VO2max for exactly the same time one to two week later. White blood cells (WBCs) were isolated by the erythrocyte lysis method. Gene expression profiles were measured using the Affymetrix GeneChipÂ® technology. After scaling, normalisation, and filtering groupwise and pairwise comparisons of gene expression intensities were performed and validated by real-time PCR. We found 450 genes up-regulated and 150 down-regulated (> 1.5-fold change; ANOVA with Benjamini-Hochberg correction for multiple testing, p<0.05) upon exhaustive exercise. Analysis of mean expression levels after MT showed that the extent of up- and down-regulation was workload dependent. The genes for the stress proteins HSPA1A, HSPH1 and the matrix metalloproteinase MMP-9 showed the most prominent increases whereas the mRNA concentrations of the YES1 oncogene (YES1), of CD160 (BY55), and of a member of the mitochondrial electron transport chain (ATP5s) were most strongly reduced. Remarkably, we could largely reproduce the data from a previous report by Connolly et al. (01) even though we used considerably different methodology. After acute exercise the genes with increased expression levels were highly significantly associated with the gene ontology terms heat-shock proteins, apoptosis and inflammation. The results demonstrate that gene expression changes in WBCs can reflect intensity and duration of exercise. Further analysis is needed to confirm the applicability of expression fingerprints as useful tools for monitoring exercise and training loads in order to avoid training associated health risks. Experiment Overall Design: Five healthy male probands executed an exhaustive treadmill test (80% VO2max) until individual exhaustion. One week later they repeated the test at 60% VO2max for the same time (moderate test). Blood samples (9ml) were drawn before and one hour past the tests. Erythrocytes were lysed and RNA was isolated from the white blood cells. RNA was processed and hybridised on U133A 2.0 Affyetrix GeneChips. Samples were grouped and gene expression changes were detected via multiple algorithms included in GeneSpring 7.2 (Agilent). Experiment Overall Design: Three groups were build from the twenty samples. All ten pre exercise samples were grouped together as the pre test" group. The other two groups contained five samples related to the "exhaustive test" or the "moderate test". TTest in cobination with multiple testing corrections, principal component analysis and hierarchical clstering were used to scan for gene expression profiles induced through the different exercise conditions.

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Transcriptome-wide microRNA expression of white blood cells before and after intense cardiorespiratory exercise

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