Project description:Unconditioned thoroughbred geldings were exercised to maximal heart rate or fatigue on an equine high-speed treadmill. Skeletal muscle biopsies were taken from the middle gluteal muscle before, immediately after and four hours after exercise. Three-condition experiment, Pre exercise (T0), Immediately post exercise (T1), 4 hours post exercise (T2). Hybridisations: T0 vs T1, T0 vs T2 Biological replicates: 8 Technical replication Dye swap
Project description:Unconditioned thoroughbred geldings were exercised to maximal heart rate or fatigue on an equine high-speed treadmill. Skeletal muscle biopsies were taken from the middle gluteal muscle before, immediately after and four hours after exercise.
Project description:Growing interest in equine regenerative biology and oxygen-dependent muscle physiology has highlighted the need for reliable in vitro models that accurately reflect the behaviour of equine skeletal muscle cells. In this study, we established an equine skeletal muscle cell line derived from post-mortem tissue. We confirmed its myogenic identity using qPCR analysis of key myogenic regulators, including MYOD1, MYF5, MYOG, and PAX7. This cell system was then used to characterise transcriptional responses to differential oxygen conditions via whole-transcriptome profiling using next-generation sequencing (NGS). By exposing cultures to normoxia and controlled hypoxia (1% and 3% O₂) for defined time periods, we aimed to investigate how oxygen availability shapes the transcriptional landscape of equine skeletal muscle cells and to provide a robust platform for future studies on muscle adaptation, metabolism, and environmental stress responses.
Project description:Digital gene expression profiling was used to characterize the assembly of genes expressed in equine skeletal muscle and to identify the subset of genes that were differentially expressed following a ten month period of exercise training. The study cohort comprised 7 thoroughbred racehorses from a single training yard. Skeletal muscle biopsies were collected at rest from the gluteus medius at two time points: T1 (unconditioned), (9 +/- 0.5 months old) and T2 (conditioned) (20 +/- 0.7 months old). The most highly abundant genes in the muscle transcriptome were those involved in muscle contraction, aerobic respiration and mitochondrial function. A previously unreported over-representation of genes relating to RNA processing, the stress response and proteolysis was observed. Following training 92 tags were differentially expressed of which 74 were annotated. Sixteen genes showed increased expression, including the mitochondrial genes, ACADVL, MRPS21 and SLC25A29. Among the 58 genes with deceased expression MSTN, a negative regulator of muscle growth had the greatest decrease. Functional analysis of all expressed genes using FatiScan revealed an asymmetric distribution of 482 Gene Ontology groups and 18 KEGG pathways. Functional groups with highly significantly (P < 0.0001) increased expression included mitochondrion, oxidative phosphorylation and fatty acid metabolism while functional groups with decreased expression were mainly associated with structural genes and included the sarcoplasm, laminin complex and cytoskeleton. Examination of muscle expression changes in 7 thoroughbred horses following 10 months of exercise training using digital gene expression with NlaIII.
Project description:The participants performed 8 weeks of superised aerobic endurance exercise. Skeletal muscle biopsise were taken at rest before and after intervention and matched analysis was performed.
Project description:Digital gene expression profiling was used to characterize the assembly of genes expressed in equine skeletal muscle and to identify the subset of genes that were differentially expressed following a ten month period of exercise training. The study cohort comprised 7 thoroughbred racehorses from a single training yard. Skeletal muscle biopsies were collected at rest from the gluteus medius at two time points: T1 (unconditioned), (9 +/- 0.5 months old) and T2 (conditioned) (20 +/- 0.7 months old). The most highly abundant genes in the muscle transcriptome were those involved in muscle contraction, aerobic respiration and mitochondrial function. A previously unreported over-representation of genes relating to RNA processing, the stress response and proteolysis was observed. Following training 92 tags were differentially expressed of which 74 were annotated. Sixteen genes showed increased expression, including the mitochondrial genes, ACADVL, MRPS21 and SLC25A29. Among the 58 genes with deceased expression MSTN, a negative regulator of muscle growth had the greatest decrease. Functional analysis of all expressed genes using FatiScan revealed an asymmetric distribution of 482 Gene Ontology groups and 18 KEGG pathways. Functional groups with highly significantly (P < 0.0001) increased expression included mitochondrion, oxidative phosphorylation and fatty acid metabolism while functional groups with decreased expression were mainly associated with structural genes and included the sarcoplasm, laminin complex and cytoskeleton.
Project description:Myofibrillar myopathy (MFM) in horses is a late onset disease that affects performance and athleticism. It is characterized by myofibrillar disarray and protein aggregation with no known cause. The objective of this study was to elucidate the molecular drivers of MFM in Warmblood (WB) horses by proteomic profiling (5 MFM WB, 4 non-MFM WB) of gluteal muscle. MFM horses used in this study had a chronic history of poor performance and exercise intolerance as well as accumulation of desmin aggregates in > 4 myofibers per muscle sample. The Equine Neuromuscular Diagnostic Laboratory database at Michigan State University was queried to identify WB horses with snap frozen gluteus medius biopsies available for analysis. Non-MFM control horses were defined as horses with no history of exercise intolerance and no evidence of desmin accumulation or other histopathology in muscle biopsies. Muscle biopsy samples were obtained at rest from horses that had not undertaken strenuous exercise in the preceding 48 hours.
Project description:Purpose: Recent research suggests that Ketone monoester (KME) supplementation may enhance skeletal muscle adaptation to exercise, possibly modulated be an increased secretion of erythropoietin (EPO) and increased muscle glycogen resynthesis, but the precise molecular pathways driving the response remain uncertain. Therefore, we aimed at characterising the modulation of skeletal muscle by KME through a genome-wide characterisation of the transcriptome early post-exercise. Methods: Following a randomized, double-blind, crossover design, recreationally active men [n= 9; age: 26 ± 5 (means ± SD); height: 1.80 ± 0.07 m; body mass: 80 ± 9 kg; V̇O2max: 47 ± 4 mL·kg-1·min-1]) completed two experimental trials where they ingested either 1.25 g/kg of KME (>96% (R)-3-hydroxybutyl (R)-3-hydroxybutyrate) or a taste-matched placebo (PLA) drink around exercise (90-min cycling at 60% of VO2max) in standardised conditions. . Venous blood samples were taken throughout baseline, exercise and recovery, and skeletal muscle biopsies were taken at baseline and 3-h post exercise. Results: KME intake elevated serum ßHB concentrations post-exercise to 3.3 ± 0.5 mM and remained elevated levels during recovery (range: ~4.0–4.7 mM). Remarkably, there was no marked upregulation or downregulation of differentially expressed genes at 3-h post-exercise with KME supplementation compared to PLA. Although, baseline serum EPO levels were higher in PLA (12.8 ± 3.9 IU/L) compared to KME (10.1 ± 5.7 IU/L) and remained slightly higher in PLA throughout recovery, there was not significant difference between conditions. Pre- and post- exercise muscle glycogen concentrate were similar between conditions (KME PRE: 501 ± 118 mmol/kg, KME POST, 284 ± 117 mmol/kg, PLA PRE: 492 ± 102 mmol/kg, PLA POST: 282 ± 184 mmol/kg). Conclusions: Our findings indicate that despite a sustained post-exercise increase in serum ßHB concentrations, KME supplementation did not induce transcriptional changes in skeletal muscle. Additionally, in contrast to previous studies, KME did not affect glycogen concentrations or serum EPO levels during the first 3-hours post-exercise.
Project description:Exercise is an important strategy in the prevention and treatment of metabolic diseases, like diabetes and obesity. Alterations in the skeletal muscle proteome, including post-translational modifications, especially acetylation, regulate its metabolic adaptations to exercise. Here, we examined the effect of 6-week aerobic exercise and Scriptaid, a HDAC4/5 inhibitor, on the proteome and acetylome of skeletal muscle in mice. We find Scriptaid and exercise both induce acetylation modification changes of some proteins involved in metabolism, suggest that exercise improves metabolic health by regulating protein acetylation level.
Project description:We aimed to investigate the human skeletal muscle (SkM) DNA methylome after exercise in low carbohydrate (CHO) energy balance (with high fat) compared with exercise in low-CHO energy deficit (with low fat) conditions. The objective to identify novel epigenetically regulated genes and pathways associated with ‘train-low sleep-low’ paradigms. The sleep-low conditions included 9 males that cycled to deplete muscle glycogen while reaching a set energy expenditure. Post-exercise, low-CHO meals (protein-matched) completely replaced (using high-fat) or only partially replaced (low-fat) the energy expended. The following morning resting baseline biopsies were taken and the participants then undertook 75 minutes of cycling exercise, with skeletal muscle biopsies collected 30 minutes and 3.5 hours post exercise. Discovery of genome-wide DNA methylation was undertaken using Illumina EPIC arrays and targeted gene expression analysis was conducted by RT-qPCR. At baseline participants under energy balance (high fat) demonstrated a predominantly hypermethylated (60%) profile across the genome compared to energy deficit-low fat conditions. However, post exercise performed in energy balance (with high fat) elicited a more prominent hypomethylation signature 30 minutes post-exercise in gene regulatory regions important for transcription (CpG islands within promoter regions) compared with exercise in energy deficit (with low fat) conditions. Such hypomethylation was enriched within pathways related to: IL6-JAK-STAT signalling, metabolic processes, p53 / cell cycle and oxidative / fatty acid metabolism. Hypomethylation within the promoter regions of genes: HDAC2, MECR, IGF2 and c13orf16 were associated with significant increases in gene expression in the post-exercise period in energy balance compared with energy deficit. Furthermore, histone deacetylase, HDAC11 was oppositely regulated at the gene expression level compared with HDAC2, where HDAC11 was hypomethylated yet increased in energy deficit compared with energy balance conditions. Overall, we identify some novel epigenetically regulated genes associated with train-low sleep-low paradigms.