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Physical exercise induces rapid release of small extracellular vesicles into the circulation.

Frühbeis C, Helmig S, Tug S, Simon P, Krämer-Albers EM - J Extracell Vesicles (2015)

Bottom Line: Here, we show that physical activity is associated with the release of nano-sized EVs into the circulation.In response to treadmill running, elevation of small EVs was moderate but appeared more sustained.We hypothesize that EVs released during physical activity may participate in cell communication during exercise-mediated adaptation processes that involve signalling across tissues and organs.

View Article: PubMed Central - PubMed

Affiliation: Molecular Cell Biology, Johannes Gutenberg-University Mainz, Mainz, Germany.

ABSTRACT
Cells secrete extracellular vesicles (EVs) by default and in response to diverse stimuli for the purpose of cell communication and tissue homeostasis. EVs are present in all body fluids including peripheral blood, and their appearance correlates with specific physiological and pathological conditions. Here, we show that physical activity is associated with the release of nano-sized EVs into the circulation. Healthy individuals were subjected to an incremental exercise protocol of cycling or running until exhaustion, and EVs were isolated from blood plasma samples taken before, immediately after and 90 min after exercise. Small EVs with the size of 100-130 nm, that carried proteins characteristic of exosomes, were significantly increased immediately after cycling exercise and declined again within 90 min at rest. In response to treadmill running, elevation of small EVs was moderate but appeared more sustained. To delineate EV release kinetics, plasma samples were additionally taken at the end of each increment of the cycling exercise protocol. Release of small EVs into the circulation was initiated in an early phase of exercise, before the individual anaerobic threshold, which is marked by the rise of lactate. Taken together, our study revealed that exercise triggers a rapid release of EVs with the characteristic size of exosomes into the circulation, initiated in the aerobic phase of exercise. We hypothesize that EVs released during physical activity may participate in cell communication during exercise-mediated adaptation processes that involve signalling across tissues and organs.

No MeSH data available.


Related in: MedlinePlus

Effect of cycling exercise on plasma small EVs. Plasma samples were collected pre, post and 90 min post (90+) ergometer exercise, and 100,000×g pellets were prepared. Particle size and concentration were measured by NTA [(a) subjects a and b, age above 30] and protein content was analyzed by Western blotting [(b) subjects 1–6]. Western blot signals of Flot1 [(c) pre/post: n=6, 90+: n=5], Hsp/Hsc70 [(d) pre/post: n=6, 90+: n=5], Tsg101 [(e) pre/post: n=3, 90+: n=2; not detectable in all experiments] and IntαIIb [(f) pre/post: n=6, 90+: n=5] were quantified (**p<0.01, Wilcoxon–Kruskal–Wallis test). Subjects older than 30 years are marked in red (subject 2, 38 years; subject 5, 58 years).
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Figure 0001: Effect of cycling exercise on plasma small EVs. Plasma samples were collected pre, post and 90 min post (90+) ergometer exercise, and 100,000×g pellets were prepared. Particle size and concentration were measured by NTA [(a) subjects a and b, age above 30] and protein content was analyzed by Western blotting [(b) subjects 1–6]. Western blot signals of Flot1 [(c) pre/post: n=6, 90+: n=5], Hsp/Hsc70 [(d) pre/post: n=6, 90+: n=5], Tsg101 [(e) pre/post: n=3, 90+: n=2; not detectable in all experiments] and IntαIIb [(f) pre/post: n=6, 90+: n=5] were quantified (**p<0.01, Wilcoxon–Kruskal–Wallis test). Subjects older than 30 years are marked in red (subject 2, 38 years; subject 5, 58 years).

Mentions: To investigate the influence of a single bout of physical activity on the levels of circulating EVs in plasma, we recruited 8 healthy, physically active men [mean (SD) age, 41.1 (±14.9) years; IAT, 181.1 (±77.8) W] performing an incremental cycling test until exhaustion [mean (SD) time 16.6 (±5) min]. We collected venous blood samples (EDTA-anticoagulated blood) before (pre), immediately after (post) and 90 min after cessation of exercise (90+) and prepared plasma. To analyse the levels of EVs in the plasma samples, we performed differential centrifugation at 10,000×g pelleting larger vesicles including platelet remnants and apoptotic bodies (here defined as crude MVs), followed by filtration of the supernatant and ultracentrifugation at 100,000×g to collect small vesicles of a size below 200 nm, which include exosomes (here defined as small EVs). Pellets of 100,000×g derived from 2 study participants (age >30 years) were analyzed by NTA revealing particles with a mean size of 120 (SEM±3.6) nm in diameter (Fig. 1a). The total amount of particles increased in average 2.7 times (SEM±0.2) directly after exercise and returned to baseline levels after 90 min. Next, we investigated 100,000×g pellets of 6 subjects by Western blotting with antibodies against the EV marker proteins Flot1, Hsp/Hsc70 and Tsg101 (Fig. 1b) and quantified signal intensities (Fig. 1c–f). The numbers of these 3 universal EV marker proteins increased on average 5.2 times (SEM±1.3) after exercise. Moreover, we detected integrin αIIb found on platelets indicating the presence of platelet-derived small EVs (Fig. 1b and f). The level of this protein rose 2.9 times (SEM±1.2) upon exercise. Samples taken in the early recovery phase after exercise (90+) revealed a decrease in the amounts of EV proteins, but the levels were not fully reaching the baseline as determined by NTA. A statistical analysis using the Wilcoxon–Kruskal–Wallis test revealed that the detected associations between release of small EVs and exercise were significant for Flot1 (p=0.0125) and Hsp70 (p=0.002) on a global level across all 3 points in time (pre, post and 90 min after exercise) with post hoc testing revealing significant increases pre to post (Flot1, p=0.0021 and Hsp/Hsc70, p=0.0021), respectively (Fig. 1c–f). Variances in the amount of small EVs released during exercise between individual subjects were noticeable and may depend on the condition of the test person and age. Individuals above the age of 30 appear to respond less intensely to exercise (n=2, highlighted by red symbols in Fig. 1b–f), although this observation certainly requires further in-depth analysis.


Physical exercise induces rapid release of small extracellular vesicles into the circulation.

Frühbeis C, Helmig S, Tug S, Simon P, Krämer-Albers EM - J Extracell Vesicles (2015)

Effect of cycling exercise on plasma small EVs. Plasma samples were collected pre, post and 90 min post (90+) ergometer exercise, and 100,000×g pellets were prepared. Particle size and concentration were measured by NTA [(a) subjects a and b, age above 30] and protein content was analyzed by Western blotting [(b) subjects 1–6]. Western blot signals of Flot1 [(c) pre/post: n=6, 90+: n=5], Hsp/Hsc70 [(d) pre/post: n=6, 90+: n=5], Tsg101 [(e) pre/post: n=3, 90+: n=2; not detectable in all experiments] and IntαIIb [(f) pre/post: n=6, 90+: n=5] were quantified (**p<0.01, Wilcoxon–Kruskal–Wallis test). Subjects older than 30 years are marked in red (subject 2, 38 years; subject 5, 58 years).
© Copyright Policy - open-access
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4491306&req=5

Figure 0001: Effect of cycling exercise on plasma small EVs. Plasma samples were collected pre, post and 90 min post (90+) ergometer exercise, and 100,000×g pellets were prepared. Particle size and concentration were measured by NTA [(a) subjects a and b, age above 30] and protein content was analyzed by Western blotting [(b) subjects 1–6]. Western blot signals of Flot1 [(c) pre/post: n=6, 90+: n=5], Hsp/Hsc70 [(d) pre/post: n=6, 90+: n=5], Tsg101 [(e) pre/post: n=3, 90+: n=2; not detectable in all experiments] and IntαIIb [(f) pre/post: n=6, 90+: n=5] were quantified (**p<0.01, Wilcoxon–Kruskal–Wallis test). Subjects older than 30 years are marked in red (subject 2, 38 years; subject 5, 58 years).
Mentions: To investigate the influence of a single bout of physical activity on the levels of circulating EVs in plasma, we recruited 8 healthy, physically active men [mean (SD) age, 41.1 (±14.9) years; IAT, 181.1 (±77.8) W] performing an incremental cycling test until exhaustion [mean (SD) time 16.6 (±5) min]. We collected venous blood samples (EDTA-anticoagulated blood) before (pre), immediately after (post) and 90 min after cessation of exercise (90+) and prepared plasma. To analyse the levels of EVs in the plasma samples, we performed differential centrifugation at 10,000×g pelleting larger vesicles including platelet remnants and apoptotic bodies (here defined as crude MVs), followed by filtration of the supernatant and ultracentrifugation at 100,000×g to collect small vesicles of a size below 200 nm, which include exosomes (here defined as small EVs). Pellets of 100,000×g derived from 2 study participants (age >30 years) were analyzed by NTA revealing particles with a mean size of 120 (SEM±3.6) nm in diameter (Fig. 1a). The total amount of particles increased in average 2.7 times (SEM±0.2) directly after exercise and returned to baseline levels after 90 min. Next, we investigated 100,000×g pellets of 6 subjects by Western blotting with antibodies against the EV marker proteins Flot1, Hsp/Hsc70 and Tsg101 (Fig. 1b) and quantified signal intensities (Fig. 1c–f). The numbers of these 3 universal EV marker proteins increased on average 5.2 times (SEM±1.3) after exercise. Moreover, we detected integrin αIIb found on platelets indicating the presence of platelet-derived small EVs (Fig. 1b and f). The level of this protein rose 2.9 times (SEM±1.2) upon exercise. Samples taken in the early recovery phase after exercise (90+) revealed a decrease in the amounts of EV proteins, but the levels were not fully reaching the baseline as determined by NTA. A statistical analysis using the Wilcoxon–Kruskal–Wallis test revealed that the detected associations between release of small EVs and exercise were significant for Flot1 (p=0.0125) and Hsp70 (p=0.002) on a global level across all 3 points in time (pre, post and 90 min after exercise) with post hoc testing revealing significant increases pre to post (Flot1, p=0.0021 and Hsp/Hsc70, p=0.0021), respectively (Fig. 1c–f). Variances in the amount of small EVs released during exercise between individual subjects were noticeable and may depend on the condition of the test person and age. Individuals above the age of 30 appear to respond less intensely to exercise (n=2, highlighted by red symbols in Fig. 1b–f), although this observation certainly requires further in-depth analysis.

Bottom Line: Here, we show that physical activity is associated with the release of nano-sized EVs into the circulation.In response to treadmill running, elevation of small EVs was moderate but appeared more sustained.We hypothesize that EVs released during physical activity may participate in cell communication during exercise-mediated adaptation processes that involve signalling across tissues and organs.

View Article: PubMed Central - PubMed

Affiliation: Molecular Cell Biology, Johannes Gutenberg-University Mainz, Mainz, Germany.

ABSTRACT
Cells secrete extracellular vesicles (EVs) by default and in response to diverse stimuli for the purpose of cell communication and tissue homeostasis. EVs are present in all body fluids including peripheral blood, and their appearance correlates with specific physiological and pathological conditions. Here, we show that physical activity is associated with the release of nano-sized EVs into the circulation. Healthy individuals were subjected to an incremental exercise protocol of cycling or running until exhaustion, and EVs were isolated from blood plasma samples taken before, immediately after and 90 min after exercise. Small EVs with the size of 100-130 nm, that carried proteins characteristic of exosomes, were significantly increased immediately after cycling exercise and declined again within 90 min at rest. In response to treadmill running, elevation of small EVs was moderate but appeared more sustained. To delineate EV release kinetics, plasma samples were additionally taken at the end of each increment of the cycling exercise protocol. Release of small EVs into the circulation was initiated in an early phase of exercise, before the individual anaerobic threshold, which is marked by the rise of lactate. Taken together, our study revealed that exercise triggers a rapid release of EVs with the characteristic size of exosomes into the circulation, initiated in the aerobic phase of exercise. We hypothesize that EVs released during physical activity may participate in cell communication during exercise-mediated adaptation processes that involve signalling across tissues and organs.

No MeSH data available.


Related in: MedlinePlus