Role of A1 receptors in renal sympathetic neurotransmission in the mouse kidney

A1 receptors may participate in renal sympathetic neurotransmission by enhancing the postjunctional effects of norepinephrine. The purpose of this study was to test this concept using A1 receptor knockout (A1AR−/−) mice. In isolated kidneys from nontransgenic mice perfused with Tyrode's solution at a constant rate, renal sympathetic nerve stimulation (RSNS) increased ( P < 0.0001) renal venous perfusate levels of inosine (adenosine metabolite) from 23.9 ± 3.7 to 32.7 ± 5.1, 68.2 ± 12.4, and 94.0 ± 14.3 ng/ml at 3, 5, and 7 Hz, respectively ( n = 28), suggesting frequency-dependent production of adenosine. Conversely, RSNS decreased ( P < 0.0001) renal venous perfusate levels of 5′-AMP (adenosine precursor) from 1.4 ± 0.3 to 1.1 ± 0.3, 0.80 ± 0.2, and 0.6 ± 0.2 ng/ml at 3, 5, and 7 Hz, respectively ( n = 28), suggesting frequency-dependent increased metabolism of 5′-AMP. In kidneys from nontransgenic mice, blockade of adenosine receptors with 1,3-dipropyl-8-p-sulfophenylxanthine attenuated ( P = 0.0130) vasoconstrictor responses to RSNS at 3, 5, and 7 Hz [control ( n = 29): 22 ± 4, 34 ± 6, 42 ± 6 mmHg, respectively; 1,3-dipropyl-8-p-sulfophenylxanthine-treated ( n = 11): 6 ± 1, 12 ± 3, 15 ± 3 mmHg, respectively]. In A1AR−/− kidneys ( n = 10), vasoconstrictor responses to RSNS at 3, 5, and 7 Hz were 7 ± 3, 20 ± 5, and 36 ± 9 mmHg, respectively. In kidneys from wild-type littermates ( n = 9), responses were 27 ± 9, 58 ± 14, and 59 ± 11 mmHg, respectively (effect of genotype: P = 0.0363). In kidneys from nontransgenic mice, 2-chloro- N6-cyclopentyladenosine (CCPA; highly selective A1 receptor agonist) increased renal vasoconstriction induced by norepinephrine ( P = 0.0008; n = 28). In kidneys from A1AR−/− the response to norepinephrine was attenuated and the ability of CCPA to enhance responses to norepinephrine was abolished. In conclusion, adenosine formed during RSNS enhances the postjunctional effects of released norepinephrine by activating A1 receptors.

Effects of exercise training on the vascular reactivity of the whole kidney circulation in rabbits

Exercise training is known to improve vasodilating mechanisms mediated by endothelium-dependent relaxing factors in the cardiac and skeletal muscle vascular beds. However, the effects of exercise training on visceral vascular reactivity, including the renal circulation, are still unclear. We used the experimental model of the isolated perfused rabbit kidney, which involves both the renal macro- and microcirculation, to test the hypothesis that exercise training improves vasodilator mechanisms in the entire renal circulation. New Zealand White rabbits were pen confined (Sed; n = 24) or treadmill trained (0% grade) for 5 days/wk at a speed of 18 m/min during 60 min over a 12-wk period (ExT; n = 24). Kidneys isolated from Sed and ExT rabbits were continuously perfused in a nonrecirculating system under conditions of constant flow and precontracted with norepinephrine (NE). We assessed the effects of exercise training on renal vascular reactivity using endothelial-dependent [acetylcholine (ACh) and bradykinin (BK)] and -independent [sodium nitroprusside (SNP)] vasodilators. ACh induced marked and dose-related vasodilator responses in kidneys from Sed rabbits, the reduction in perfusion pressure reaching 41 ± 8% ( n = 6; P < 0.05). In the kidneys from ExT rabbits, vasodilation induced by ACh was significantly enhanced to 54 ± 6% ( n = 6; P < 0.05). In contrast, BK-induced renal vasodilation was not enhanced by training [19 ± 8 and 13 ± 4% reduction in perfusion pressure for Sed and ExT rabbits, respectively ( n = 6; P > 0.05)]. Continuous perfusion of isolated kidneys from ExT animals with Nω-nitro-l-arginine methyl ester (l-NAME; 300 μM), an inhibitor of nitric oxide (NO) biosynthesis, completely blunted the additional vasodilation elicited by ACh [reduction in perfusion pressure of 54 ± 6 and 38 ± 5% for ExT and l-NAME + ExT, respectively ( n = 6; P < 0.05)]. On the other hand, l-NAME infusion did not affect ACh-induced vasodilation in Sed animals. Exercise training also increased renal vasodilation induced by SNP [36 ± 7 and 45 ± 10% reduction in perfusion pressure for Sed and ExT rabbits, respectively ( n = 6; P < 0.05)]. It is concluded that exercise training alters the rabbit kidney vascular reactivity, enhancing endothelium-dependent and -independent renal vasodilation. This effect seems to be related not only to an increased bioavailability of NO but also to the enhanced responsiveness of the renal vascular smooth muscle to NO.

Mechanisms involved in the activation of ischemically sensitive, afferent renal nerve mediated reflex increases in hind-limb vascular resistance in the anesthetized rabbit

Acute occlusion of the renal circulation in the anesthetized rabbit results in a neurally mediated, reflex increase in hind-limb vascular resistance, which is flow rather than pressure dependent. This suggests that the activating stimulus could be ischemia. In the present study vascularly isolated kidneys were perfused with hypoxemic or hypercapnic blood, and the hind-limb vascular response was measured. Renal perfusion with hypoxemic blood resulted in an increase in femoral perfusion pressure (FPP), which was negatively correlated with the oxygen tension of the blood. At a [Formula: see text] of 36.4 ± 0.9 mmHg (1 mmHg = 133.3 Pa), FPP rose by 34.4 ± 5.7 mmHg. Renal denervation abolished this effect. Renal perfusion with hypercapnic blood had no effect on FPP. Prostaglandin E2, bradykinin, and adenosine are released during renal ischemia and have been implicated in the mediation of afferent renal nerve activity; intrarenal administration (prostaglandin E2, 10 μg; bradykinin, 5 μg; adenosine, 20 μg; as a 1-mL bolus) during renal perfusion with normoxemic blood elicited increases in FPP of 32.4 ± 13.2, 19.2 ± 3.7, and 55.6 ± 17.8 mmHg, respectively. Intrarenal indomethacin, aprotonin, and aminophylline all inhibited the increase in FPP observed during renal perfusion with hypoxemic blood. These data indicate that renal hypoxemia stimulates an afferent renal nerve mediated increase in FPP and suggest that prostaglandin E2, bradykinin, and adenosine may all be involved in the activation of ischemically sensitive R1 chemoreceptors.Key words: renal ischemia, chemoreceptor, prostaglandins, bradykinin, adenosine.

Mechanisms for altered endothelium-dependent vasorelaxation in isolated kidneys from experimental hypertensive rats

To study mechanisms for attenuated endothelium-dependent vasorelaxation in hypertension, we examined the effects of acetylcholine (ACh) on renal vascular resistance (RVR) and release rates of endothelium-derived relaxing factor (EDRF) in kidneys isolated from spontaneously hypertensive rats (SHR), deoxycorticosterone acetate (DOCA) salt-hypertensive (DOCA salt) rats, and Dahl salt-sensitive (Dahl S) rats. Decreases in RVR by ACh were smaller in hypertensive rats than in their normotensive controls. The release rate of nitric oxide into the perfusate, which was estimated using nitrite-nitrate as an index, did not differ between SHR and Wistar-Kyoto rats (WKY). However, the release rate of EDRF was markedly decreased in both DOCA salt rats and Dahl S rats compared with their normotensive controls (10(-7) M ACh: DOCA salt 45 +/- 6 vs. control 410 +/- 60 pmol.min-1.g-1 kidney wt, P < 0.001). In SHR, high-K+ perfusion or pretreatment with glibenclamide, inhibitors of endothelium-derived hyperpolarizing factor (EDHF), significantly reduced ACh-induced vasorelaxation only in WKY, resulting in no differences in the RVR reduction between SHR and WKY. Thus attenuated ACh-induced vasorelaxation in the SHR kidney may be attributed to a decrease in EDHF, but to a decrease in EDRF in DOCA salt rats and Dahl S rats.