Figure 1. 15-20 mins of swimming exercise (Ex) decreases native human
x237A;-synuclein protein species: A. 66-<480 kDa range proteins in worms exercised for 15-20 min. on Day 1 of adulthood are significantly reduced compared to control (un-treated) and 15-20 min food restricted (FR) worms (***p≤0.0001, **p≤0.0088). These proteins are also significantly reduced for food restricted versus control (un-treated) worms (*p≤0.0116.) B. Representative Blue Native (BN) page western blot showing the protein profile of control (un-treated), Ex, and FR worms, demonstrating a reduction in proteins in 66-<480 kDa, 720 kDa, and well proteins. C. 720 kDa proteins in Ex worms are significantly reduced compared to control (un-treated) worms and almost significantly reduced compared to FR worms (***p≤0.0008, p=0.0548.) 720 kDa proteins are not significantly reduced in FR worms (p=0.4301). D. Well proteins in Ex worms are significantly reduced compared to control (un-treated) and FR worms (***p≤0.0002, ****p<0.0001). Well proteins are also significantly reduced in FR worms as compared to control (un-treated) worms (*p≤0.0144). N= 8, n=25 per replicate for A-D. E. Thrashing assay results on Day 2 show that worms previously exercised for 15-20 min on Day 1 have a significantly higher thrashing speed at 3 min compared with control (un-treated) (*p≤0.0250) or FR worms (**p≤0.0027). F. Thrashing assay results on Day 2 at 20 mins show that control (un-treated) worms have attained a higher thrashing rate, matching that of Ex worms and are significantly faster than FR worms (****p<0.0001). Ex worms maintain approximately the same thrashing speed as at 3 mins, which is also significantly higher than for FR worms (***p≤0.0003). FR worms do not see an increase in thrashing speed from 3 mins to 20 mins. G. Thrashing assay results on Day 4 demonstrate that at 3 mins, all three treatment groups display approximately the same lower thrashing speed as compared to Day 2 results. H. Thrashing assay results on Day 4 at 20 mins show that Ex worms have a significantly higher thrashing rate than control (un-treated) worms (*p≤0.0371) and an almost significant trend towards a higher thrashing rate compared with FR worms (p=0.0673). All worms show an improvement in thrashing rate compared to their speed at 3 mins; Ex worms show an average thrashing speed similar to that of control (un-treated) worms on Day 2. N=3, n=~25 worms per replicate for E-H. I-N. Representative confocal images of
x237A;-synuclein puncta (green) after background subtraction was applied for I. control (un-treated) worms; K. Ex worms; and M. FR worms, and with calculated Imaris surfaces (pink) for J. control (un-treated) worms; L. Ex worms; N. FR worms. O. Heat map showing individual surface areas (SA's) for each puncta for each worm across all biological replicates normalized to respective controls (right Y-axis). Total puncta count for all worms combined is shown on left Y-axis. Exercise shows a higher concentration of smaller sized puncta shown in dark blue (smaller than 2 units as seen in legend.)