Effects of Voluntary Running Wheel Activity and Hypertension on the Brain of Female Spontaneously Hypertensive Rats (SHRs)

Figures

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  Figure 1. Effect of free voluntary wheel activity on heart rate and blood pressure. (A) Heart rate (HR) is given for normotensive Wistar rats (WIS; n = 4), spontaneously hypertensive rats held under standard (sedentary) conditions (SHR-S; n = 9), and spontaneously hypertensive rats with free access to running wheels (SHR-R; n = 9). (B) Systolic blood pressure (Psyst) and (C) diastolic blood pressure (Pdiast) is given for the same rats. Data are given as means ± S.D. with original data points as dots. Statistical comparison was done by one-way ANOVA with p = 0.000225 for HR, p = 0.000004 for Psyst, and p = 0.000002 for Pdiast with Student–Newman–Keul’s post hoc analysis. *; p < 0.05 vs. WIS; #, p < 0.05 vs. SHR-S.

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  Figure 2. Venn diagram indicating the number of differentially regulated genes (DRGs) in the three brain areas that show a correlation with blood pressure. Of note, one gene (Ucp2) is differentially expressed in all three parts of the brain.

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  Figure 3. Correlation between blood pressure and Ucp2 expression. (A) Black circles indicate data from WIS (n = 4), blue circles indicate data from SHR-S (n = 9), and red circles indicate data from SHR-R (n = 9). p values are 0.015 (cortex; r2 = 0.406), 0.001 (medulla oblongata; r2 = 0.440), and p < 0.000 for olfactory bulb (r2 = 0.669). (B) Western blots indicating the UCP2 protein expression in the different areas from the brain. (C) Quantification of the Western blots. Data are given as means ± S.D. with original data points as dots. WIS (n = 3); SHR-S (n = 4); SHR-R (n = 4) *, p < 0.05 vs. WIS.

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  Figure 4. Group comparison of the gene expression of (A) UCP2, (B) CAT, and (C) REN in the three different parts of the brain. Data are means ± S.D. and the expression in WIS (n = 10) was set as 1.0 and compared with SHR-S (n = 21) and SHR-R (n = 15). Statistical comparison was done by one-way ANOVA with p values of p < 0.000 (Ucp2, cortex), p < 0.000 (Ucp2, med. obl.), p < 0.000 (Ucp2, olf. bulb), p = 0.001 (Cat, med. obl.), p < 0.000 (Cat, olf. bulb); and p < 0.000 (Ren, med. obl.). All other comparisons with p > 0.005. Student–Newman–Keul’s post hoc analysis was performed. Data are given as means ± S.D. with original data points as dots. *, p < 0.05 vs. WIS.

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  Figure 5. Venn diagram indicating the number of differentially regulated genes (DRGs) in the three brain areas that show a correlation with running activity (km/w). Of note, none of the genes were differentially expressed in all three parts of the brain.

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  Figure 6. Correlation between running activity and gene expression in the cortex (n = 15). p values are 0.009 (A, Ucp3; r2 = 0.412), 0.016 (B, Ace1; r2 = 0.369), 0.036 (C, Vegfa; r2 = 0.295), 0.020 (D, App; r2 = 0.350), and 0.043 (E, Nse; r2 = 0.280).

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  Figure 7. Group comparison of endothelin-1 expression in the three different parts of the brain. (A) Data are means ± S.D. and the expression in WIS (n = 10) was set as 1.0 and compared with SHR-S (n = 21) and SHR-R (n = 15). Statistical comparison was done by one-way ANOVA with p values of p < 0.05. (B) Western blots indicating the expression of endothelin-1. (C) Quantification of the Western blots (WIS, n = 3; SHR-S; n = 4; SHR-R, n = 4). Statistical comparison was done using a Kruskal–Wallis test with p values of 0.056 (cortex) and 0.069 (olf. bulb). t test differences with p < 0.05 are indicated by asterisks. Data are given as means ± S.D. with original data points as dots.

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  Figure 8. Visualization of oxidative stress by DHE staining. Cortical slices were stained. (A) Representative original images of four rats. (B) Means ± S.D. of these slices. Statistical comparison was done by one-way ANOVA with p values of p < 0.000. Student–Newman–Keul’s post hoc analysis was performed. *; p < 0.05 vs. WIS; #; p < 0.05 vs. SHR-S. Data are given as means ± S.D. with original data points as dots.

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  Figure 9. Group comparison of BACE1 expression in the three different parts of the brain. (A) Data are means ± S.D. and the expression in WIS (n = 10) was set as 1.0 and compared with SHR-S (n = 21) and SHR-R (n = 15). Statistical comparison was done by one-way ANOVA with p values of p > 0.05 for all groups. (B) Western blots indicating the expression of BACE1. (C) Quantification of the Western blots. Statistical comparison was done using one-way ANOVA with Student–Newman–Keuls post hoc analysis. *; p < 0.05 vs. WIS. Data are given as means ± S.D. with original data points as dots.

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  Figure 10. Brain preparation from rats showing the position of the tissues used in this study.