Integrated mRNA and miRNA expression profiling in blood reveals candidate biomarkers associated with endurance exercise in horse (mRNA)
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ABSTRACT: The adaptive response to extreme endurance exercise might involve transcriptional and translational regulation by microRNAs (miRNAs). Therefore, the aim of this study was to define an integrative analysis of blood transcriptome and miRNome in horses before and after a long endurance ride (160 km) using equine microarrays. A total of 2,453 genes and 162 miRNAs were found to be differentially expressed (DEG) between animals at rest and after the endurance ride. To gain understanding of the biological functions regulated by the differentially expressed miRNA, we used a hypergeometric test analysis. Notably, we detected 42 differentially expressed miRNAs that putatively regulate a total of 350 depleted DEGs, involved in glucose metabolism, fatty acid oxidation, mitochondrion biogenesis, and immune response pathways. Graphical Gaussian models in an independent validation set of animals confirmed that 4 miRNAs could be strong candidate regulatory molecules for endurance exercise adaptation. This study represents, to the best of our knowledge, the first integrated comprehensive overview of the miRNA-mRNA co-regulation networks that may play a central role in controlling post-transcriptomic regulations during endurance exercise in horses.
Project description:The adaptive response to extreme endurance exercise might involve transcriptional and translational regulation by microRNAs (miRNAs). Therefore, the aim of this study was to define an integrative analysis of blood transcriptome and miRNome in horses before and after a long endurance ride (160 km) using equine microarrays. A total of 2,453 genes and 162 miRNAs were found to be differentially expressed (DEG) between animals at rest and after the endurance ride. To gain understanding of the biological functions regulated by the differentially expressed miRNA, we used a hypergeometric test analysis. Notably, we detected 42 differentially expressed miRNAs that putatively regulate a total of 350 depleted DEGs, involved in glucose metabolism, fatty acid oxidation, mitochondrion biogenesis, and immune response pathways. Graphical Gaussian models in an independent validation set of animals confirmed that 4 miRNAs could be strong candidate regulatory molecules for endurance exercise adaptation. This study represents, to the best of our knowledge, the first integrated comprehensive overview of the miRNA-mRNA co-regulation networks that may play a central role in controlling post-transcriptomic regulations during endurance exercise in horses. Sixty-one Arabian or half-breed Arabian horses (20 females and 41 geldings) aged 10 ± 2 years (±SEM) were recruited on voluntary basis of the owner on three 160 km endurance rides.
Project description:The adaptive response to extreme endurance exercise might involve transcriptional and translational regulation by microRNAs (miRNAs). Therefore, the aim of this study was to define an integrative analysis of blood transcriptome and miRNome in horses before and after a long endurance ride (160 km) using equine microarrays. A total of 2,453 genes and 162 miRNAs were found to be differentially expressed (DEG) between animals at rest and after the endurance ride. To gain understanding of the biological functions regulated by the differentially expressed miRNA, we used a hypergeometric test analysis. Notably, we detected 42 differentially expressed miRNAs that putatively regulate a total of 350 depleted DEGs, involved in glucose metabolism, fatty acid oxidation, mitochondrion biogenesis, and immune response pathways. Graphical Gaussian models in an independent validation set of animals confirmed that 4 miRNAs could be strong candidate regulatory molecules for endurance exercise adaptation. This study represents, to the best of our knowledge, the first integrated comprehensive overview of the miRNA-mRNA co-regulation networks that may play a central role in controlling post-transcriptomic regulations during endurance exercise in horses. Sixty-one Arabian or half-breed Arabian horses (20 females and 41 geldings) aged 10 ± 2 years (±SEM) were recruited on voluntary basis of the owner on three 160 km endurance rides.
Project description:The adaptive response to extreme endurance exercise might involve transcriptional and translational regulation by microRNAs (miRNAs). Therefore, the aim of this study was to define an integrative analysis of blood transcriptome and miRNome in horses before and after a long endurance ride (160 km) using equine microarrays. A total of 2,453 genes and 162 miRNAs were found to be differentially expressed (DEG) between animals at rest and after the endurance ride. To gain understanding of the biological functions regulated by the differentially expressed miRNA, we used a hypergeometric test analysis. Notably, we detected 42 differentially expressed miRNAs that putatively regulate a total of 350 depleted DEGs, involved in glucose metabolism, fatty acid oxidation, mitochondrion biogenesis, and immune response pathways. Graphical Gaussian models in an independent validation set of animals confirmed that 4 miRNAs could be strong candidate regulatory molecules for endurance exercise adaptation. This study represents, to the best of our knowledge, the first integrated comprehensive overview of the miRNA-mRNA co-regulation networks that may play a central role in controlling post-transcriptomic regulations during endurance exercise in horses.
Project description:Endurance exercise in horses implies adaptive processes involving affective, physiological, biochemical, and cognitive-behavioral response in an attempt to regain homeostasis. We hypothesized that the identification of the relationships between blood metabolome, transcriptome and miRNome during endurance exercise could provide significant insights into the molecular response to intense exercise or prediction of this response at basal status. In this perspective, the serum metabolome and whole-blood transcriptome and miRNome data were obtained from 10 horses before and after a 160 km endurance competition. Results: We obtained a global regulatory network based on 11 unique metabolites, 263 metabolic genes and 5 miRNAs whose expression was significantly altered at T1 (post- endurance competition) relative to T0 (baseline, pre- endurance competition). This network provided new insights into the cross talk between the distinct molecular pathways (e.g. energy and oxygen sensing, oxidative stress, and inflammation) that were not detectable when analyzing single metabolites or transcripts alone. This suggested that single metabolites and transcripts were carrying out multiple roles and thus sharing several biochemical pathways. Using a regulatory impact factor metric analysis, this regulatory network was further confirmed at the transcription factor and miRNA levels. In an extended cohort of 39 animals, multiple factor analysis confirmed the strong associations between lactate, methylene derivatives, miR-21-5p, miR-16-5p, and genes that coded proteins involved in metabolic reactions primarily related to energy, ubiquitin proteasome and lipopolysaccharide immune responses at T1. Multiple factorial analyses also identified potential biomarkers at T0 for an increased possibility of failure to finish an endurance competition.
Project description:A single bout of exercise induces changes in gene expression in skeletal muscle. Regular exercise results in an adaptive response involving changes in muscle architecture and biochemistry, and is an effective way to manage and prevent common human diseases such as obesity, cardiovascular disorders and type II diabetes. Our study is a transcriptome-wide analysis of skeletal muscle tissue in a large cohort of untrained Thoroughbred horses before and after a bout of high-intensity exercise and again after an extended period of training. We hypothesized that regular high-intensity exercise training primes the transcriptome for the demands of high-intensity exercise.
Project description:The molecular mechanisms of exercise-induced cardiovascular protection are poorly understood. There is growing evidence that reactive oxygen species (ROS) are necessary for some of these adaptations and antioxidants may be used to investigate this effect. This study aimed to determine the effects of exercise and/or antioxidant supplementation on myocardial and vascular endothelium gene expression. Male Wistar rats were divided into four groups: i) endurance exercise (90min of treadmill running 4d/week, 14 weeks); ii) antioxidant-treated; iii) antioxidant and endurance exercise and iv) control. The supplemented animals received Vitamin E (1000 IU/kg diet) and ï¡-lipoic acid (1.6 g/kg diet) mixed with rat chow. cDNA microarray analysis was performed using purified endothelial RNA from myocardial and coronary artery endothelial cells and showed that the expression levels of 35, 40 and 40 genes were altered for groups i, ii, and iii respectively compared to control. Differentially expressed genes were analysed using the KEGG pathway database, hierarchical cluster and DAVID analysis. These analyses revealed that a gene involved in cardiovascular disease progression, Ras homolog gene family member A (RhoA) was down-regulated by exercise, upregulated by antioxidant supplementation and the combination of exercise and antioxidant blunted both effects. These findings were confirmed by real-time PCR. In summary, exercise and antioxidant supplementation affect endothelial cell gene expression and ROS appear necessary for some of these adaptations. 48 Male Wistar rats were divided into four groups: i) endurance exercise (90min of treadmill running 4d/week, 14 weeks); ii) antioxidant-treated; iii) antioxidant and endurance exercise and iv) control. The supplemented animals received Vitamin E (1000 IU/kg diet) and ï¡-lipoic acid (1.6 g/kg diet) mixed with rat chow. cDNA microarray analysis was performed using purified endothelial RNA from myocardial and coronary artery endothelial cells. Based on the limited material available, it was necessary to combine the RNA into three pools per treatment group for CAEC (approx 400 ng of total RNA), and four pools for LVEC (approx 1 μg of total RNA) with an additional reference pool from each control group.
Project description:Capacity of exercise and performance is the most valuable in the horses. They have been selected for strength, speed, and indurance trait. Athletic pheno types are influenced markedly by environment, management, and training. However, it has long been accepted that there are underlying genetic factors. To determine altered mRNA expression in circulating leukocytes of horses induced by exercise. Healthy neutered male warmblood horses were subjected to indoor exercise (trotting with alternative cantering for 6o minutes). Peripheral blood was collected from the jugular vein before and after the exercise, and subsequently buffy coat leukocytes were isolated by centrifugation. Total RNAs was isolated. Cyanine 3-labeled cRNA (complementary RNA) was generated from Agilentâs Low RNA Input Linear Amplification kit with 500 ng total RNA. Labeled cRNA was applied microarray (Agilent technologies, 8x60K) using Agilentâs Gene Expression Hybridization Kit. The present study revealed a subset of mRNAs in equine peripheral blood leukocytes affected by exercise, providing background information for genes associated with exercise in warm-blood horses. Three healthy, gelding warmblood horses between 9 and 17 yr were selected. 6 samples were collected containing 3 samples before exercise and 3 samples after exercise
Project description:Endurance exercise has a dramatic impact on the functionality of mitochondria and on the composition of intestinal microbiome, but the mechanisms regulating the crosstalk between these two components are still largely unknown. Here, we sampled 20 elite horses before and after an endurance race and used blood transcriptome, blood metabolome and fecal microbiome to describe the microbiota-mitochondria inter-talk. A subset of mitochondria-related differentially expressed genes involved in pathways such as energy metabolism, oxidative stress and inflammation was discovered and then shown to be associated with butyrate-producing bacteria of the Lachnospiraceae family, especially Eubacterium. The mechanisms involved were not fully understood, but through the action of their metabolites likely acted on PPARδ, the FRX-CREB axis and their downstream targets to delay the onset of hypoglycemia, inflammation and extend running time. Our results also suggested that circulating free fatty acids may act not merely as fuel but drive mitochondrial inflammatory responses triggered by the translocation of gut bacterial polysaccharides following endurance. Targeting the gut-mitochondria axis appears therefore as a potential strategy to enhance athletic performance.
Project description:Objective: The objective of this study was to characterize extracellular vesicles (EVs) in plasma and synovial fluid obtained from horses with and without naturally occurring post-traumatic osteoarthritis (PTOA). Animals (Samples): EVs were isolated from plasma and synovial fluid from horses with (n = 6) and without (n = 6) PTOA. Methods: Plasma and synovial fluid EVs were characterized with respect to quantity, size, and surface markers. Small RNA sequencing was performed and differentially expressed miRNAs underwent bioinformatic analysis to identify putative targets and to explore potential associations with specific biological processes. Results: Plasma and synovial fluid samples from horses with PTOA had a significantly higher proportion of exosomes and a lower proportion of microvesicles compared to horses without PTOA. Small RNA sequencing revealed several differentially expressed miRNAs including miR-144, miR-219-3p, and miR-199a-3p in plasma and miR-199a-3p, miR-214, and miR-9094 in synovial fluid EVs. Bioinformatics analysis of the differentially expressed miRNAs highlighted their potential role in fibrosis, differentiation of chondrocytes, apoptosis, and inflammation pathways in PTOA. Clinical Relevance: We have identified dynamic molecular changes in small non-coding signatures of plasma and synovial fluid EVs in horses with naturally occurring PTOA. These findings could serve to identify promising biomarkers in the pathogenesis of PTOA, to facilitate the development of targeted therapies, and to aid in establishing appropriate translational models of PTOA.
Project description:Capacity of exercise and performance is the most valuable in the horses. They have been selected for strength, speed, and indurance trait. Athletic pheno types are influenced markedly by environment, management, and training. However, it has long been accepted that there are underlying genetic factors. To determine altered mRNA expression in circulating leukocytes of horses induced by exercise. Healthy neutered male warmblood horses were subjected to indoor exercise (trotting with alternative cantering for 6o minutes). Peripheral blood was collected from the jugular vein before and after the exercise, and subsequently buffy coat leukocytes were isolated by centrifugation. Total RNAs was isolated. Cyanine 3-labeled cRNA (complementary RNA) was generated from Agilent’s Low RNA Input Linear Amplification kit with 500 ng total RNA. Labeled cRNA was applied microarray (Agilent technologies, 8x60K) using Agilent’s Gene Expression Hybridization Kit. The present study revealed a subset of mRNAs in equine peripheral blood leukocytes affected by exercise, providing background information for genes associated with exercise in warm-blood horses.