Disruption of the with no lysine kinase–STE20-proline alanine-rich kinase pathway reduces the hypertension induced by angiotensin II

Figures

• 

  FIGURE 1. The arterial hypertension induced by angiotensin II infusion is blunted in Ste20-related proline–alanine-rich kinase-knock-in mice. (a) SBP and (b) mean blood pressure. Graphs represent SBP and mean blood pressure average of 3-day 1-h measures before angiotensin II infusion (basal) and the average of 12th, 13th and 14th-day 1-h after angiotensin II infusion (14-day) of wild-type (white bars) and Ste20-related proline–alanine-rich kinase-knock-in (black bars) mice. Blood pressure was assessed using radiotelemetry every day from 100 to 1100 h, during 17 days (before and after angiotensin II infusion). Each bar represents the mean ± standard error of approximately 550 measures of each mouse. n = 5 mice for each group. ∗P less than 0.05 wild-type vs. Ste20-related proline–alanine-rich kinase-knock-in.

• 

  FIGURE 2. Angiotensin II infusion induced an increase in Na–Cl cotransporter expression and phosphorylation in wild-type, but not in Ste20-related proline–alanine-rich kinase-knock-in mice. (a) Representative western blot for total Na–Cl cotransporter, phosphor-Na–Cl cotransporter and β-Actin from proteins extracted from the kidney of wild-type or Ste20-related proline–alanine-rich kinase-knock-in mice infused with vehicle or angiotensin II, as stated. (b) Densitometric analysis of two independent western blot for total Na–Cl cotransporter and phosphor-Na–Cl cotransporter expressed as the pNa–Cl cotransporter/total Na–Cl cotransporter ratio after β-actin normalization. Open bars indicate vehicle infusion and closed bars indicate angiotensin II infusion. ∗P less than 0.0001 vs. all other groups.

• 

  FIGURE 3. The effect in blood pressure induced by aldosterone infusion was higher in Ste20-related proline–alanine-rich kinase-knock-in mice. (a) SBP and (b) mean blood pressure. Graphs represent SBP and mean blood pressure average of 3-day 1-h measures before aldosterone infusion (basal) and the average of 12th, 13th and 14th-day 1-h after aldosterone infusion (14-day) of wild type (white bars) and Ste20-related proline–alanine-rich kinase-knock-in (black bars) mice. Blood pressure was assessed using radiotelemetry every day from 1000 to 1100 h, during 17 days (before and after aldosterone infusion). Each bar represents the mean ± standard error of approximately 550 measures of each mouse of each day. n = 5 mice for each group. ∗P less than 0.05 vs. Ste20-related proline–alanine-rich kinase-knock-in.

• 

  FIGURE 4. Aldosterone infusion increased the pNa–Cl cotransporter/Na–Cl cotransporter ratio in wild-type mice, but not in Ste20-related proline–alanine-rich kinase-knock-in mice. (a) Representative western blot for total Na–Cl cotransporter, phosphor-Na–Cl cotransporter and β-actin of proteins extracted from the kidney of wild-type or Ste20-related proline–alanine-rich kinase-knock-in mice infused with vehicle or Aldosterone, as stated. (b) Densitometric analysis of two independent western blot for total Na–Cl cotransporter and phosphor-Na–Cl cotransporter, expressed as the pNa–Cl cotransporter/total Na–Cl cotransporter ratio after β-actin normalization. Open bars indicate vehicle infusion, and closed bars indicate aldosterone infusion. (c) Plasma potassium levels after 13-day treatment with aldosterone or angiotensin II in STE20-proline alanine-rich kinase-WT and Ste20-related proline–alanine-rich kinase-knock-in mice. n = 5. Open bars show STE20-proline alanine-rich kinase-WT mice and close bars show Ste20-related proline–alanine-rich kinase-knock-in mice. ∗P less than 0.05 vs. Ste20-related proline–alanine-rich kinase-knock-in basal.

• 

  FIGURE 5. Diuretic challenge with hydrochlorothiazide or amiloride in STE20-proline alanine-rich kinase-WT and Ste20-related proline–alanine-rich kinase-knock-in mice infused with aldosterone and 1% NaCl in the drinking water. Mice were kept in a metabolic cage to determine the basal urinary sodium excretion (adjusted per volume as mEq sodium over mg of creatinine in urine) and the urinary sodium excretion after a single i.p. injection of hydrochlorothiazide (50 mg/kg) or amiloride (5 mg/kg). Bars represent mean ± standard error in wild-type (white) and Ste20-related proline–alanine-rich kinase-knock-in (black), before (R0), 4 and 20 h after amiloride administration, and 6 and 18 h after hydrochlorothiazide administration. n = 5. ∗P less than 0.05 vs. Ste20-related proline–alanine-rich kinase-knock-in at 4, 6 and 18 h.

• 

  FIGURE 6. Effect of amiloride injection in blood pressure in aldosterone-infused and 1% Na–Cl Ste20-related proline–alanine-rich kinase-knock-in and STE20-proline alanine-rich kinase-WT mice. Closed squares indicate wild-type mice, and open circles indicate Ste20-related proline–alanine-rich kinase-knock-in mice. By telemetric measured, the response to a single doses of amiloride was tested on 1% Na–Cl-aldosterone-infused Ste20-related proline–alanine-rich kinase-knock-in and STE20-proline alanine-rich kinase. The blood pressure was recorded continuously 2 h before and 12 h after amiloride administration. Each point represents the mean ± standard error of approximately 550 measurements equivalent to 1 h of recording for each mouse. n = 5 mice from each group. ∗P less than 0.05 vs. Ste20-related proline–alanine-rich kinase-knock-in.