Impaired myogenic autoregulation in kidneys of Brown Norway rats.

The Brown Norway (BN) rat is normotensive and has an extended lifespan but is extremely sensitive to hypertension-induced renal injury. Relative impairment of autoregulation has been implicated in the progression of renal failure whereas absence of myogenic autoregulation is associated with early renal failure. Therefore, we tested the hypothesis that there is conditional failure of renal autoregulation in BN rats. In isoflurane-anesthetized BN rats, the pressure-flow transfer function was normal when pressure fluctuated spontaneously. External forcing increased pressure fluctuation and exposed weakness of the myogenic component of autoregulation; the component mediated by tubuloglomerular feedback was less affected. In the presence of vasopressin to raise renal perfusion pressure, myogenic autoregulation was further impaired during forcing in BN rats but not in Wistar rats. Compensation by the myogenic system was rapidly restored on cessation of forcing, suggesting a functional limitation rather than a structural failure. Graded forcing in Wistar rats and in spontaneously hypertensive rats revealed that compensation due to the myogenic system was strong and independent of forcing amplitude. In contrast, graded forcing in BN rats showed that compensation was reduced when fluctuation of blood pressure was increased but that the reduction was independent of forcing amplitude. The results demonstrate conditional failure of myogenic autoregulation in BN rats. These acute studies provide a possible explanation for the observed sensitivity to hypertension-induced renal injury in BN rats.

Responses of mesenteric and renal blood flow dynamics to acute denervation in anesthetized rats.

Previous studies have shown that renal autoregulation dynamically stabilizes renal blood flow (RBF). The role of renal nerves, particularly of a baroreflex component, in dynamic regulation of RBF remains unclear. The relative roles of autoregulation and mesenteric nerves in dynamic regulation of blood flow in the superior mesenteric artery (MBF) are similarly unclear. In this study, transfer function analysis was used to identify autoregulatory and baroreflex components in the dynamic regulation of RBF and MBF in Wistar rats and young spontaneously hypertensive rats (SHR) anesthetized with isoflurane or halothane. Wistar rats showed effective dynamic autoregulation of both MBF and RBF, as did SHR. Autoregulation was faster in the kidney (0.22 +/- 0.01 Hz) than in the gut (0.13 +/- 0.01 Hz). In the mesenteric, but not the renal bed, the admittance phase was significantly negative between 0.25 and 0. 7 Hz, and the negative phase was abrogated by mesenteric denervation, indicating the presence of an arterial baroreflex. The baroreflex was faster than autoregulation in either bed. The presence of sympathetic effects unrelated to blood pressure was inferred in both vascular beds and appeared to be stronger in the SHR than in the Wistar rats. It is concluded that a physiologically significant baroreflex operates on the mesenteric, but not the renal circulation and that blood flow in both beds is effectively stabilized by autoregulation.

Pharmacological modulation of spontaneous renal blood flow dynamics.

Two mechanisms contribute to renal autoregulation. The faster system, which is thought to be myogenic, operates at 0.1-0.2 Hz (i.e., 5-10 s/cycle), while the slower one, tubuloglomerular feedback, operates at 0.03-0.05 Hz (i.e., 20-30 s/cycle). Both attenuate spontaneous or induced fluctuations of blood pressure, but it has proven difficult to separate their individual contributions because there is potential for interaction between the two. The present study was designed to examine the dynamics of the faster system during pharmacological blockade of tubuloglomerular feedback. Normotensive and hypertensive rats were studied under isoflurane or halothane anesthesia. Administration of the loop diuretic furosemide plus the angiotensin II (ANGII) AT1 receptor antagonist losartan caused a 10-fold or greater natriuresis, indicating profound inhibition of ascending limb salt transport, and also produced characteristic changes in the transfer function relating blood pressure (input) to renal blood flow (output). Operation of the 0.1-0.2 Hz mechanism was essentially unaltered, as shown by the presence of a peak in phase angle at 0.1-0.2 Hz and reduction of gain at frequencies slower than 0.15 Hz. The 0.03-0.05 Hz mechanism was markedly inhibited, as shown by loss of the second phase angle peak at 0.03-0.05 Hz, loss of the local maximum in gain at 0.05 Hz, and loss of the second gain reduction below 0.05 Hz. Both during control and after inhibition of tubuloglomerular feedback, the 0.1-0.2 Hz system attenuated = 50% of the effects of spontaneous blood pressure fluctuations, suggesting that this mechanism, operating alone, can significantly stabilize renal blood flow in the face of spontaneous fluctuations of blood pressure.

Alpha 2-adrenergic mediation of the effects of angiotensin II on rat renal artery in vitro.

Angiotensin II (ANG II) is a major influence on renal blood flow, acting directly on the renal vasculature and upon other controllers. In vivo observations suggest that ANG II affects renal artery resistance, although explicit in vitro studies have produced negative results. To resolve this issue, potential interactive effects of ANG II on renal artery in vitro were tested. Renal arteries were harvested from ketamine-anesthetized Sprague-Dawley rats and perfused in vitro at constant flow. Resistance was determined from the axial pressure drop while downstream pressure was held constant at approximately 80 mmHg (1 mmHg = 133.3 Pa). ANG II, per se, had only a trivial effect on arterial diameter (-5.5 +/- 1.5% at 10(-7) M ANG II) and failed to affect resistance at concentrations ranging from 10(-11) to 10(-7) M. Norepinephrine caused strong concentration-dependent constriction, increasing resistance from 0.32 +/- 0.04 to 1.86 +/- 0.73 mmHg.mL-1.min at 10(-6) M and 3.27 +/- 0.88 mmHg.mL-1.min at 10(-5) M. In the presence of 10(-8) M ANG II, these responses were significantly increased to 3.31 +/- 1.00 and 5.02 +/- 1.22 mmHg.mL-1.min, respectively. Similarly, in the presence of 10(-6) M norepinephrine, ANG II caused significant, concentration-dependent constriction of renal artery. In a separate experiment, 10(-7) M yohimbine, a relatively specific antagonist of alpha 2-adrenergic receptors, reversed the resistance increment due to ANG II, but not that due to norepinephrine. When yohimbine was applied before norepinephrine and ANG II, it did not affect the response to norepinephrine, but again blocked potentiation of the response by ANG II.(ABSTRACT TRUNCATED AT 250 WORDS)

In vitro response of rat renal artery to perfusion pressure.

Both tubuloglomerular feedback and a myogenic response contribute to autoregulation of renal blood flow. Vascular interaction initiated by tubuloglomerular feedback has been described and prevents definition, in vivo, of the contribution of myogenic responses to autoregulation. Segments of rat renal artery were perfused in vitro at constant flow while upstream and downstream pressures were measured on-line, allowing determination of resistance. Transmural pressure was governed by a downstream resistor. Outside diameter was measured by an ocular micrometer. The segments were bathed in bicarbonate Ringer solution and perfused with Ringer containing 50 g/L bovine serum albumin. Potassium depolarization reduced the diameter and made it more sensitive to perfusion pressure. Serosal norepinephrine, 10(-7)-10(-5) M, caused graded constriction and increased the axial pressure drop due to vessel resistance. Addition to the perfusate of rat red blood cells to hematocrit approximately 33% significantly reduced arterial diameter and enhanced the increased axial pressure drop induced by 10(-6) M norepinephrine. Sequential elevation of perfusion pressure from 50 to 100 mmHg (1 mmHg = 133.3 Pa) increased the diameter significantly. Red cells reduced the slope of the diameter-pressure relationship. In another experiment, norepinephrine reduced the slope of diameter versus perfusion pressure, while 10(-4) M papaverine plus norepinephrine increased the slope, compared with norepinephrine alone. Norepinephrine caused a sizable axial pressure drop (15.7 +/- 3.7 mmHg), which decayed as perfusion pressure increased; the decay was accentuated by papaverine. The changes in axial pressure drop were linearly related to the inverse 4th power of diameter, indicating that both measurements assessed the same behavior. Several different maneuvers thus affect the relationship between arterial diameter and perfusion pressure, and the relationship between axial pressure drop and perfusion pressure. The results indicate the presence of a myogenic response, which is, however, not strong enough to defend vessel diameter when pressure rises.

Inhibition of alpha-adrenergic responses in the rat liver by lipophilic K+ channel blockers or depolarizing Cl- gradients. Evidence for a potential-sensitive step in the signal transduction path.

To study the role of K+ channels and membrane potential in alpha-adrenergic responses of the rat liver, lipophilic K+ channel blockers quinidine and 4-aminopyridine were used or external Cl- was replaced with gluconate, an impermeant ion. Glucose release, O2 uptake, portal pressure, and K+ flux were measured in the isolated perfused liver. The alpha-agonist phenylephrine caused biphasic changes in each parameter, a fast transient followed by sustained elevated responses. Infusion of 5 mM 4-aminopyridine, 0.1 mM quinidine, or gluconate prior to phenylephrine inhibited each parameter, with the greatest inhibition occurring during the second phase. A similar pattern was seen with 2 mM EGTA. This contrasts with the full inhibition of all responses following exposure to the alpha-antagonist phentolamine. Infusion of each inhibitor at the peak of the sustained phase inhibited all responses. Phenylephrine-stimulated release of K+ was augmented in the presence of EGTA and was inhibited by 4-aminopyridine or quinidine. In contrast, beta-adrenergic stimulation of glucose release and K+ flux were not affected by the K+ channel blockers. Phenylephrine-stimulated glucose release from hepatocyte suspensions decreased by about 50% in the presence of 4-aminopyridine, EGTA, or gluconate. The results are discussed in terms of a potential role for K+ channels in alpha-adrenergic signal transduction in the liver.

Induction of haemodynamic oscillations in the perfused rat liver by K+ channel blockers.

1. Exposure of the isolated perfused (constant flow) rat liver to the K+ channel blockers 4-aminopyridine (4-AP) or Cs+ causes the appearance of oscillations in portal pressure and oxygen uptake. The oscillations have a mean frequency of 0.035 Hz (2.1 cycles/min) and are fully reversible upon perfusion with blocker-free saline. Tetraethylammonium (0.17-24.7 mM) does not induce oscillatory behaviour. 2. Reversible block of the 4-AP-induced oscillations is caused by 2 mM-EGTA, or verapamil, chlorpheniramine, phentolamine or propranolol with IC50 values of 0.42, 13.5, 15 or 11.5 microM respectively. The oscillations are transiently blocked by atropine (IC50 = 8.3 microM at peak inhibition) and are not affected by 2.7 microM-tetrodotoxin. 3. Endothelium-dependent vasorelaxants, Kupffer cell activity modifiers, retrograde perfusion, or removal of the portal vein from the circuit do not modify the oscillation parameters. 4. Oscillations are also caused by infusion of physiological concentrations of adrenaline or phenylephrine, but not isoprenaline. 5. The results provide new evidence for the existence of intrahepatic voltage-sensitive Ca2+, and 4-AP- and Cs(+)-sensitive K+ channels. We propose that the K+ channel blockers reveal an intrinsic oscillator in the liver, and that phasic vasoactivity may involve a minor contribution from neurotransmitter and/or hormonal substances.