Single-cell transcriptomic analyses unveil exercise-induced changes in murine skin vascular endothelial cells
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ABSTRACT:
PROVIDER: PRJNA1256714 | ENA |
REPOSITORIES: ENA
Dataset's files
| Action | DRS | |||
|---|---|---|---|---|
| SRR33366271_1.fastq.gz | Fastqsanger.gz | |||
| SRR33366271_2.fastq.gz | Fastqsanger.gz | |||
| SRR33366272_1.fastq.gz | Fastqsanger.gz | |||
| SRR33366272_2.fastq.gz | Fastqsanger.gz | |||
| SRR33366273_1.fastq.gz | Fastqsanger.gz |
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Project description:Physical activity (PA) is a fundamental aspect of preventive medicine, offering profound benefits for cardiovascular health and overall well-being. Despite its widespread benefits, the molecular mechanisms underlying PA-induced improvements in microvascular functions remain poorly understood. The skin microvasculature is uniquely affected by exercise-induced shear stress, especially during thermoregulation. Using single-cell RNA sequencing, we investigated how voluntary exercise influences the transcription profile of endothelial cells in the skin microvasculature of mice. We assigned 20 mice to either a sedentary group or a 1-month voluntary exercise program involving running wheels. Post-intervention, we collected skin biopsies from twelve mice for transcriptomic analyses. The differential expression analysis showed a significant increase in the expression of the Zbtb16 gene in vascular endothelial cells (vECs). The expression of Zbtb16 in vECs was correlated with genes involved in angiogenesis. Additionally, Gene Set Enrichment Analysis (GSEA) with nominally differentially expressed genes in vECs highlighted the suppression of pathways related to oxidative stress, cell proliferation and metabolism in the exercise group. This suggests an exercise-triggered transition towards improved metabolic efficiency and enhanced homeostasis in vECs. These results begin to reveal how exercise induces molecular changes in vECs of the skin microvasculature, highlighting PA's role in influencing endothelial function.
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Project description:Exercise improves cardiometabolic and vascular function, though mechanisms remain unclear. Our objective was to demonstrate the diversity of circulating extracellular RNA (ex-RNA) release during acute exercise in humans and its relevance to exercise-mediated benefits on vascular inflammation. We performed plasma small RNA sequencing (RNA-seq) in 26 individuals undergoing symptom-limited maximal treadmill exercise, with replication of our top candidate miRNA in a separate cohort of 59 individuals undergoing bicycle ergometry. We found changes in miRNAs and other ex-RNAs with exercise (e.g., y-RNAs, t-RNAs) implicated in cardiovascular disease. In two independent cohorts of acute maximal exercise, we identified miR-181b-5p as a key ex-RNA increased in plasma after exercise, with validation in a separate cohort. In a mouse model of acute exercise, we found significant increases in miR-181b-5p expression in skeletal muscle after acute exercise in young (but not older) mice. Previous work revealed a strong role for miR-181b-5p in vascular inflammation in obesity, insulin resistance, sepsis, and cardiovascular disease. Circulating ex-RNAs altered in plasma after acute exercise target pathways involved in inflammation, including miR-181b-5p. Further investigation into the role of known (e.g., miRNA) and novel (e.g., y-RNAs) is warranted to uncover new mechanisms of vascular inflammation on exercise-mediated benefits on health.
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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.
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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 alpha-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.
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Project description:Exercise is a powerful driver of physiological angiogenesis during adulthood, but the mechanisms of exercise-induced vascular expansion are poorly understood. We explored endothelial heterogeneity in skeletal muscle and identified two capillary muscle endothelial cells (mEC) populations which are characterized by differential expression of ATF3/4. Spatial mapping showed that ATF3/4 + mECs are enriched in red oxidative muscle areas while ATF3/4 low ECs lie adjacent to white glycolytic fibers. In vitro and in vivo experiments revealed that red ATF3/4 + mECs are more angiogenic when compared to white ATF3/4 low mECs. Mechanistically, ATF3/4 in mECs control genes involved in amino acid uptake and metabolism and metabolically prime red (ATF3/4 + ) mECs for angiogenesis. As a consequence, supplementation of non-essential amino acids and overexpression of ATF4 increased proliferation of white mECs. Finally, deleting Atf4 in ECs impaired exercise-induced angiogenesis. Our findings illustrate that spatial metabolic angiodiversity determines the angiogenic potential of muscle ECs.
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Project description:The prevalence and mortality rates of type 2 diabetes mellitus (T2DM) continue to increase, imposing a significant burden on individuals and nations worldwide. The pancreas plays an important role in T2DM development, while exercise is acrucial nonpharmacological treatment. Although the pancreas comprises various cell types, current studies on the effects of exercise on diabetes have mainly focused on the beta cells. In this study, we aimed to enhance our understanding of the effects of exercise on other pancreatic cell types. This was aimed at facilitating the comprehensive analysis of the processes and principles by which exercise protects and enhances pancreatic function. Six-week-old male db/db mice were trained on a treadmill at a speed of 9 m/min for 10 weeks (50 min/day, 5 days/week). Single-cell RNA sequencing (scRNA-seq) was performed to analyze cell types in the pancreas. The results showed that exercise improved the body weight and pancreas profile of db/db mice.The scRNA-seq demonstrated that the pancreas was composed of 12 cell types, with obvious changes in endothelial cells (ECs) among all groups. Further subtype analysis suggested that ECs could be annotated into five subtypes, with capillary ECs presenting remarkable differences among the groups. Additionally, Gene Ontology (GO) enrichment analysis showed the close role of capillary ECs, which are a cell group involved in vascular development and regulation of EC proliferation and migration in blood vessels. The newly identified capillary ECs may be involved in the protective effect of exercise against pancreatic injury in db/db mice.
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