Contribution of Orexin to the Neurogenic Hypertension in BPH/2J Mice.

BPH/2J mice are a genetic model of hypertension associated with an overactive sympathetic nervous system. Orexin is a neuropeptide which influences sympathetic activity and blood pressure. Orexin precursor mRNA expression is greater in hypothalamic tissue of BPH/2J compared with normotensive BPN/3J mice. To determine whether enhanced orexinergic signaling contributes to the hypertension, BPH/2J and BPN/3J mice were preimplanted with radiotelemetry probes to compare blood pressure 1 hour before and 5 hours after administration of almorexant, an orexin receptor antagonist. Mid frequency mean arterial pressure power and the depressor response to ganglion blockade were also used as indicators of sympathetic nervous system activity. Administration of almorexant at 100 (IP) and 300 mg/kg (oral) in BPH/2J mice during the dark-active period (2 hours after lights off) markedly reduced blood pressure (-16.1 ± 1.6 and -11.0 ± 1.1 mm Hg, respectively;P<0.001 compared with vehicle). However, when almorexant (100 mg/kg, IP) was administered during the light-inactive period (5 hours before lights off) no reduction from baseline was observed (P=0.64). The same dose of almorexant in BPN/3J mice had no effect on blood pressure during the dark (P=0.79) or light periods (P=0.24). Almorexant attenuated the depressor response to ganglion blockade (P=0.018) and reduced the mid frequency mean arterial pressure power in BPH/2J mice (P<0.001), but not BPN/3J mice (P=0.70). Immunohistochemical labeling revealed that BPH/2J mice have 29% more orexin neurons than BPN/3J mice which are preferentially located in the lateral hypothalamus. The results suggest that enhanced orexinergic signaling contributes to sympathetic overactivity and hypertension during the dark period in BPH/2J mice.

Comparison of sympathetic nerve activity normalization procedures in conscious rabbits.

One of the main constraints associated with recording sympathetic nerve activity (SNA) in both humans and experimental animals is that microvolt values reflect characteristics of the recording conditions and limit comparisons between different experimental groups. The nasopharyngeal response has been validated for normalizing renal SNA (RSNA) in conscious rabbits, and in humans muscle SNA is normalized to the maximum burst in the resting period. We compared these two methods of normalization to determine whether either could detect elevated RSNA in hypertensive rabbits compared with normotensive controls. We also tested whether either method eliminated differences based only on different recording conditions by separating RSNA of control (sham) rabbits into two groups with low or high microvolts. Hypertension was induced by 5 wk of renal clipping (2K1C), 3 wk of high-fat diet (HFD), or 3 mo infusion of a low dose of angiotensin (ANG II). Normalization to the nasopharyngeal response revealed RSNA that was 88, 51, and 34% greater in 2K1C, HFD, and ANG II rabbits, respectively, than shams (P < 0.05), but normalization to the maximum burst showed no differences. The RSNA baroreflex followed a similar pattern whether RSNA was expressed in microvolts or normalized. Both methods abolished the difference between low and high microvolt RSNA. These results suggest that maximum burst amplitude is a useful technique for minimizing differences between recording conditions but is unable to detect real differences between groups. We conclude that the nasopharyngeal reflex is the superior method for normalizing sympathetic recordings in conscious rabbits.

Differential activation of renal sympathetic burst amplitude and frequency during hypoxia, stress and baroreflexes with chronic angiotensin treatment.

NEW FINDINGS: What is the central question of this study? Is the elevated tonic renal nerve activity induced by chronic angiotensin administration mediated by recruitment or increased firing frequency and does this occur via stress, chemoreflex or baroreflex pathways? What is the main finding and its importance? Long-term angiotensin treatment in rabbits elevates renal sympathetic nerve activity by recruitment of previously silent fibres. This was similar to the effect of chemoreflex stimulation, but not to stress or baroreceptor activation, suggesting that presympathetic pathways activated by angiotensin may be common to those activated by chemoreceptors. Modulation of sympathetic nerve activity involves control by the CNS of the amplitude of neural discharges, reflecting recruitment of neurons and their firing frequency. We tested whether elevated tonic renal sympathetic nerve activity (RSNA) induced by chronic angiotensin administration is mediated by recruitment or increased firing frequency and whether this is characteristic of the pattern observed with activation of stress, chemoreflex or baroreflex pathways. Conscious rabbits treated with angiotensin II for 12 weeks to increase blood pressure by 10-30% were subjected to stress (air jet), hypoxia (10% O2 + 3% CO2) and drug-induced changes in blood pressure to produce baroreflexes. Total RSNA and RSNA burst amplitude were scaled to 100 normalized units (n.u.) by the maximal response to smoke. After 12 weeks of treatment, blood pressure was 17% higher than baseline 68 ± 1 mmHg (P = 0.02). Compared with sham treatment, total RSNA and burst amplitude were +82% (P < 0.001) and 39% (P = 0.04) greater, but burst frequency was similar. Total RSNA increased during hypoxia (+38% from 4.9 ± 0.7 n.u.), owing to greater amplitude, but not frequency. Air-jet stress increased total RSNA (+44% from 4.3 ± 0.5 n.u.) and burst frequency (+21% from 5.4 ± 0.7 bursts s(-1) ), but not amplitude. Angiotensin enhanced total RSNA responses to both air jet (+33%) and hypoxia (+58%), but only increased the amplitude response to air jet. The RSNA baroreflexes reset to the higher blood pressure, but amplitude or frequency was not differentially altered. Chronic angiotensin treatment elevated RSNA by recruitment of neurons, which is similar to chemoreflex stimulation, but not to stress or baroreceptor activation, suggesting that presympathetic pathways activated by angiotensin may be common to those activated by chemoreceptors.

Renal sympathetic activation from long-term low-dose angiotensin II infusion in rabbits.

OBJECTIVE: Activation of renal sympathetic nerve activity (RSNA) has not been observed during long-term infusion of angiotensin II (AngII) which results in marked hypertension, despite activation of hypothalamic autonomic regions. We examined whether the function of central pathways influencing sympathetic activity is altered in conscious rabbits given a low dose of AngII that produces a modest hypertension and, therefore, limited secondary complications. METHODS: Rabbits received AngII (20-30 ng/kg per min, subcutaneously) or sham treatment for 3 months at which time they were implanted with a renal sympathetic nerve electrode and the responses to airjet stress, baroreflexes and hypoxia were examined. RESULTS: AngII infusion for 3 months increased mean arterial pressure by 16% and RSNA by 43%. Increases in RSNA during airjet stress and hypoxia (10% O2) were 35 and 65% greater in AngII-treated rabbits than sham controls, respectively. Tachycardic responses were also enhanced. Baroreflexes were shifted to the right and upward in the AngII animals but baroreflex gain was similar in the two groups, indicating near complete resetting. Greater neuronal Fos-related antigen immunoreactivity was found in the vascular organ of the lamina terminalis, paraventricular and supraoptic hypothalamic nuclei in AngII-treated rabbits compared with sham. CONCLUSION: Our results suggest that low-dose AngII-treatment results in marked sympathetic activation at rest and during stress and hypoxia, due to activation of specific hypothalamic pathways. These mechanisms may contribute to sympathetic activation in conditions associated with chronic activation of the renin-angiotensin system such as obesity or renovascular disease.

Enhanced responses to ganglion blockade do not reflect sympathetic nervous system contribution to angiotensin II-induced hypertension.

OBJECTIVE: We examined whether a specific increase in sympathetic nervous system (SNS) activity accounts for the enhanced depressor response to ganglion blockade in angiotensin II (AngII)-induced hypertension in rabbits or whether it reflects a general increased sensitivity of arterial pressure to vasodilatation. METHODS: Rabbits were renal denervated or sham-operated and 2 weeks later AngII (50 ng/kg per min) infusion commenced. Mean arterial pressure (MAP) responses to ganglion blockade (pentolinium) and vasodilators nitroprusside and adenosine were measured 2-4 weeks later. RESULTS: Basal MAP was 74 +/- 2 mmHg and maximum hypotensive responses to pentolinium, nitroprusside and adenosine were -17 +/- 2, -17 +/- 1 and -21 +/- 2 mmHg. AngII increased MAP similarly in intact and renal denervated rabbits (+25 +/- 4 mmHg and +31 +/- 4 mmHg, respectively). In intact rabbits, depressor responses to pentolinium were augmented by 75% during AngII infusion but responses to vasodilators also increased by 73-106% suggesting general augmentation of vascular reactivity rather than a specific increase in SNS neural activity. Consistent with this notion, total noradrenaline spillover was similar in normal and AngII-treated rabbits. In renal denervated rabbits, AngII enhanced depressor responses to vasodilators but not pentolinium, suggesting that sympathetic activity may be reduced by AngII hypertension when renal nerves are absent. In anaesthetized rabbits, methoxamine-induced decreases in hindlimb vascular conductance were greater in hypertensive than normotensive rabbits suggesting the presence of vascular hypertrophy of sufficient magnitude to explain increased responses to ganglion blockade and vasodilators. CONCLUSION: Enhanced depressor responses to ganglion blockade in AngII hypertension do not reflect augmented SNS activity, but rather, augmented sympathetic vasoconstriction mediated by a vascular amplifier effect.

Levels of renal and extrarenal sympathetic drive in angiotensin II-induced hypertension.

We examined the contribution of the renal nerves to mean arterial pressure (MAP) during 5-week chronic infusion of angiotensin II (Ang II; 50 ng/kg per minute SC) in conscious rabbits. Basal MAP was 68+/-1 mm Hg, and the maximum depressor response to ganglion blockade was -20+/-2 mm Hg. MAP increased by 25+/-2 mm Hg after 1 week and remained stable over the next 4 weeks. Depressor responses to pentolinium (6 mg/kg IV) were similar to control during the first week of hypertension but thereafter became increasingly greater in Ang II-treated rabbits but not vehicle-treated rabbits. After 5 weeks, the fall in MAP was 54% greater in Ang II- than in vehicle-treated rabbits (-34+/-2 versus -22+/-2 mm Hg), but renal sympathetic nerve activity was similar in both groups. Renal denervation produced a small fall in MAP in all of the vehicle-treated rabbits after 4 days (-6+/-2 mm Hg; P=0.01), but there was no consistent effect in hypertensive rabbits. The depressor response to ganglion blockade was enhanced in vehicle-treated but not Ang II-treated rabbits. The finding that renal sympathetic nerve activity is not altered by Ang II hypertension nor is the hypertension altered by renal denervation suggests that renal sympathetic nerves do not contribute to the hypertension. The greater depressor effect of acute ganglion blockade in hypertensive rabbits suggests that the sympathetic nervous system exerts increased vasoconstriction in the peripheral vasculature in Ang II-induced hypertension.