Renal Na excretion in dehydrated and rehydrated adrenalectomized sheep maintained with aldosterone.

The effect of water deprivation for 19 h on renal Na excretion of conscious adrenalectomized (ADX) sheep maintained on a constant intravenous infusion of aldosterone and cortisol (ADX-constant steroid sheep) was investigated. Both ADX and normal sheep showed large increases in renal Na excretion when they were deprived of water. ADX-constant steroid sheep also exhibited a normal postprandial natriuresis 3-6 h after feeding, whether or not water was available to drink. In another experiment, sheep deprived of water for 41 h were then allowed to drink water. Both normal and ADX-constant steroid sheep exhibited a large reduction of renal Na excretion in the 6 h after rehydration. Changes in plasma Na and K concentration and osmolality were similar in normal and ADX-constant steroid sheep during periods of dehydration and rehydration. These results show that change in aldosterone secretion is not a major factor in causing either dehydration-induced or postprandial natriuresis. Neither is it a major cause of rehydration-induced renal Na retention.

Central losartan blocks natriuretic, vasopressin, and pressor responses to central hypertonic NaCl in sheep.

This study investigated the effect of intracerebroventricular administration of the angiotensin AT1 receptor antagonist losartan on the natriuresis, pressor effect, and arginine vasopressin (AVP) secretion caused by intracerebroventricular infusion of either ANG II, hypertonic saline, or carbachol. Losartan (1 mg/h) or artificial cerebrospinal fluid (CSF) was infused into the lateral ventricle before, during, and after infusions of either ANG II at 10 microg/h for 1 h, 0.75 mol/l NaCl at 50 microl/min for 20 min, or carbachol at 1.66 microg/min for 15 min. Intracerebroventricular infusions of ANG II, 0.75 mol/l NaCl, or carbachol caused increases in renal Na+ and K+ excretion, arterial pressure, and plasma AVP levels. Increases in arterial pressure, Na+ excretion, and plasma AVP concentration ([AVP]) in response to intracerebroventricular ANG II or intracerebroventricular 0.75 mol/l NaCl were either abolished or attenuated by intracerebroventricular infusion of losartan but not by intracerebroventricular infusion of artificial CSF or intravenous losartan. Intracerebroventricular losartan did not reduce the increase in plasma [AVP] or arterial pressure in response to intracerebroventricular carbachol, but it did attenuate the natriuretic response to intracerebroventricular carbachol. We conclude that an intracerebroventricular dose of losartan (1 mg/h) that inhibits responses to intracerebroventricular ANG II also inhibits vasopressin secretion, natriuresis, and the pressor response to intracerebroventricular hypertonic saline. These results suggest that common neural pathways are involved in the responses induced by intracerebroventricular administration of ANG II and intracerebroventricular hypertonic NaCl. We propose that intracerebroventricular infusion of hypertonic saline activates angiotensinergic pathways in the central nervous system subserving the regulation of fluid and electrolyte balance and arterial pressure in sheep.

The role of angiotensin in arterial blood pressure regulation in the toad Bufo marinus.

Little is known about the role of the renin-angiotensin system in anuran amphibians, although they appear to possess the functional components of such a system. We investigated the role of angiotensin (ANG) in arterial blood pressure regulation in the conscious toad Bufo marinus using the angiotensin-converting enzyme blocker captopril. We found that conversion of endogenous ANG I to ANG II made a significant contribution to mean arterial pressure in undisturbed animals. The vascular tone contributed by ANG II was not mediated via &agr ; adrenergic mechanisms because increases in pressure in response to ANG infusion were unaffected by the presence of the &agr ; antagonist phentolamine. Angiotensin-induced vasoconstriction was shown to be an important mechanism in arterial blood pressure regulation in the face of an acute hypotensive perturbation of pressure brought about by sodium nitroprusside. Blockade of the conversion of ANG I to ANG II significantly delayed the recovery of mean arterial pressure after sodium nitroprusside-induced hypotension. This suggests that the renin-angiotensin system may play an important role in the initial responses to hypotension in anurans, whether brought about by haemorrhage or dehydration.

Intracerebroventricular losartan inhibits postprandial drinking in sheep.

We investigated the contribution of brain angiotensinergic mechanisms to postprandial drinking in sheep. Sheep in fluid balance were given 0.8 kg chaff for 30 min, and water intake was measured for the next hour. Intracerebroventricular infusion of the AT1 type angiotensin II (ANG II) receptor blocker losartan (1 mg/h) reduced postprandial drinking by approximately 70% (n = 7, P < 0.01) but did not affect food intake. The same losartan dose given intravenously had little or no effect on prandial drinking. Feeding increased Na+ concentrations in plasma and cerebrospinal fluid (CSF; n = 5, P < 0.05). Intracerebroventricular losartan (1 mg/h) inhibited the drinking responses to intracarotid infusion of ANG II (0.8 microg/min for 30 min, n = 4, P < 0.01) and to intracerebroventricular infusion of 0.5 M NaCl (1 ml/h for 1 h, n = 5, P < 0.05) but had no effect on drinking responses to intravenous infusion of 4 M NaCl (1.3 ml/min for 30 min). These findings indicate that a brain ANG II-dependent mechanism is involved in postprandial drinking in sheep. They suggest also that the mechanism by which increasing CSF Na+ causes thirst involves brain ANG II and is different from the mechanism subserving the drinking response to changes in blood Na+.

Effects of central losartan on plasma renin and centrally mediated natriuresis.

Because intracerebroventricular (ICV) infusion of hypertonic saline or angiotensin II (Ang II) both induce water drinking, vasopressin secretion, natriuresis and increased arterial pressure, the possibility that common neural pathways mediate responses to ICV Ang II and hypertonic saline has been investigated. This was done by testing the effect of ICV infusion of the Ang II antagonist losartan on the natriuretic and pressor responses to ICV hypertonic NaCl in sheep. The effect of ICV losartan on plasma renin concentration (PRC) was also investigated. Infusion of losartan (1 mg/hr) into a lateral ventricle prevented both natriuretic and pressor responses to infusion of 0.6 mol/liter NaCl into a lateral ventricle at 1 ml/hr. In another experiment, ICV losartan at 1 mg/hr caused a pronounced increase in the PRC of Na-depleted sheep, while ICV Ang II at 3 micrograms/hr decreased PRC. The results suggest that: (i) a central angiotensinergic pathway may mediate osmoregulatory responses to centrally administered hypertonic saline, and (ii) a central angiotensinergic pathway may have a tonic inhibitory influence on renin secretion in Na-depleted animals.

Schedule-induced polydipsia in rats with gastric fistulas.

We have investigated the development and maintenance of schedule-induced polydipsia (SIP) when ingested water (and food) was allowed to drain from the stomach. Fourteen male Long-Evans rats were prepared with permanent gastric cannulas and, after recovery, their body weight was reduced to 80%. Water intake was measured, with cannulas open or closed, during 42 daily 1-h sessions in which 45-mg food pellets were delivered one per minute. Allowing ingested material to drain from the stomach impaired the development of SIP and reduced the polydipsia in rats in which SIP had already been established. In contrast, opening the gastric fistulas increased drinking in these rats when all the food pellets were provided at once or after water deprivation. The opposite effects of gastric drainage on dehydration and schedule-induced drinking is consistent with the view that SIP is not a fluid-regulating phenomenon. It is not clear, however, how these unexpected findings fit current hypotheses to explain SIP that are based on oral, neural excitatory, or emotional mechanisms.

Stimulation of drinking by bacterial endotoxins in the rat.

We investigated the effects of endotoxins on water balance, rectal temperature, and food intake in male Long-Evans rats with femoral venous catheters. Extracts of Escherichia coli or Salmonella minnesota, in doses ranging from 125 to 500 micrograms/kg IV, stimulated drinking and reduced urinary water loss for several hours. The net gain of 5 ml water 2 h after the lowest dose of E. coli endotoxin was sufficient to reduce plasma osmolality and sodium concentration 2 to 3%. Drinking occurred during the period of hypothermia that frequently precedes the onset of endotoxin-induced fever, so cannot be attributed to increased body temperature. Doses of endotoxin causing drinking inhibited both spontaneous and deprivation-induced feeding. The cause of the drinking is not known, but may involve mechanisms other than the known dehydrational signals controlling thirst.

Endotoxin stimulates drinking in rats without changing dehydrational signals controlling thirst.

Intravenous injections of endotoxins from Escherichia coli or Salmonella minnesota stimulate drinking and reduce urinary excretion of water and solutes in rats. E. coli endotoxin (0.15 or 0.45 mg/kg i.v.) stimulated drinking without increasing plasma osmolality or sodium concentration, hematocrit, blood hemoglobin, or plasma protein concentration and without decreasing arterial pressure. Similarly, a dipsogenic dose of S. minnesota endotoxin (0.25 mg/kg i.v.) did not reduce arterial or venous pressures or change heart rate. Blocking the renin-angiotensin system with captopril or blocking histamine receptors with pyrilamine and cimetidine did not reduce drinking or urinary fluid retention caused by E. coli endotoxin. Injections of 10 or 450 ng E. coli endotoxin into a lateral cerebral ventricle increased body temperature but not water intake. In contrast to its stimulatory effect in water-replete rats, E. coli endotoxin (0.45 mg/kg i.v.) inhibited drinking in 24-h water-deprived rats. Thus we find no evidence to support the hypothesis that endotoxin causes thirst by changing known physiological signals of cellular or extracellular dehydration. The mechanism remains unknown.

Water and solute balance in rats during 10 h water deprivation and rehydration.

Rats with bladder and venous cannulas were deprived of water from midnight (00:00) to 10:00. Water deprivation reduced food intake within 2 h, reducing the amount of water sequestered in the gut and the solute load to the tissues. There was little change in either urinary water loss or osmolality, but water-deprived rats excreted more Na+, K+, and Cl- than food-matched controls. The change in solute balance helped preserve osmolality and cell volume at the expense of extracellular fluid volume. When water was returned, rats quickly drank enough to restore the intracellular but not the extracellular fluid deficit. Plasma osmolality and sodium concentration fell below predeprivation values. Urine osmolality and excretion of Na+, K+, and Cl- fell rapidly after drinking. Drinking continued at a slower rate for at least 4 h, but urine flow also increased so water balance stabilized. The changes in intake and electrolyte excretion during water deprivation and rehydration illustrate the important role of changes in solute balance in fluid homeostasis.

Investigating the role of angiotensin II in thirst: interactions between arterial pressure and the control of drinking.

Several lines of evidence suggest that angiotensin II plays a physiological role in the control of thirst. Establishing that, however, has been surprisingly difficult, given our current knowledge about the renin-angiotensin systems in the circulation and the brain and the variety of techniques available to measure and manipulate them. A major problem is that stimulating or blocking the renin-angiotensin system affects several physiological variables simultaneously. Since several of these variables also influence the controls of water intake directly or indirectly, the interpretation of the effect on drinking becomes more difficult. To illustrate the problem and recent developments, this paper describes some of the interactions between the effects of angiotensin II on arterial pressure and thirst, and it shows how they have contributed to the controversy over the physiological role of the peptide.

Relationship between thirst and diazoxide-induced hypotension in rats.

Diazoxide, a potent vasodilator and antidiuretic, was used to examine the relationship between hypotension and thirst in conscious rats with indwelling arterial and venous catheters. Bolus iv. injections (5-50 mg/kg) caused prompt, long-lasting, and dose-dependent reductions in mean arterial pressure (MAP) and stimulated drinking. Water intake and degree of hypotension were closely correlated when MAP was 10-65 mmHg below normal. At the time of drinking there were no significant changes in central venous pressure, plasma osmolality, or Na+ or K+ concentration. Plasma glucose increased approximately 35%, and blood volume increased approximately 10% (based on hematocrit changes and dilution of Evans blue). Captopril (100 mg/kg sc to block the renin-angiotensin system) enhanced the depressor response to diazoxide but abolished the dipsogenic response over the same range of arterial pressures tested in controls. Angiotensin II iv infusion restored drinking in captopril-treated animals. The combination of captopril and diazoxide did not block drinking to iv infusions of hypertonic saline or water deprivation. These results confirm that hypotension potently stimulates thirst and support the hypothesis that angiotensin II mediates the dipsogenic response in rats.

Acute non-haemorrhagic conjunctivitis due to coxsackievirus A24.

Two cases of acute non-haemorrhagic conjunctivitis due to coxsackievirus A24 (CA24) are described. These are the first Australian isolates. The presentation was as a severe conjunctivitis in otherwise healthy adults who had not travelled outside Australia. The course was of short duration and self-limiting with no long-term sequelae. The isolates could not be neutralised by antiserum prepared against prototype CA24 but were identified by immune electron microscopy and complement fixation.

Test of a criterion for selecting intracranial doses of angiotensin receptor blockers.

Investigators using intracerebroventricular (ICV) injections of competitive antagonists of angiotensin II (Ang II) to study thirst usually select doses sufficient to block drinking to IV Ang II. We questioned whether this test truly indicates the dose needed under physiological conditions when Ang II-induced hypertension, which inhibits thirst, is not present. Rats were prepared with chronic venous and ICV cannulas, plus femoral arterial cannulas in those used to measure arterial pressure. Captopril (100 mg/kg SC) was given before all experiments to block endogenous Ang II production. The test dose of Ang II, 50 ng/kg/min IV for 1 hr, increased water intake and arterial pressure. We selected an ICV dose of saralasin (Sar1Ala8Ang II), 4 micrograms bolus and 4 micrograms/hr for 75 min, that did not stimulate drinking itself and completely blocked drinking to IV Ang II. This dose of saralasin only partially (45%) reduced drinking to the same dose of Ang II IV when arterial pressure was lowered by giving the vasodilator diazoxide (15 mg/kg IV). Diazoxide itself did not stimulate drinking. These results support our concern that the criterion normally used to select ICV doses of Ang II antagonists probably underestimates the amount needed to inhibit angiotensinergic drinking in hypovolemic or hypotensive animals.

Effect of arterial pressure on drinking and urinary responses to angiotensin II.

To investigate the relationship between angiotensin II (ANG II) and mean arterial pressure (MAP) in the control of drinking in rats, we infused ANG II intravenously at constant rates (either 50 or 100 ng.kg-1.min-1 for 90 min) and varied MAP by intravenous injections of diazoxide (5-20 mg/kg). Rats were pretreated with captopril to block the endogenous synthesis of ANG II. When given alone, low and high doses of ANG II increased MAP approximately 30 and 50 mmHg, respectively. The low but not the high dose significantly increased water intake above control levels. Both doses caused such a large diuresis and natriuresis that the net effect was fluid loss. Reducing MAP toward normal greatly increased the drinking response to the high but not the low dose of ANG II and reduced the urinary solute and water loss to both doses. These results support the hypothesis that water intake and net fluid gain are inhibited when MAP is above normal. When MAP was reduced below normal in rats given constant infusions of ANG II the amount of water drunk and net fluid gain was proportional to the dose of ANG II but not the dose of diazoxide, the degree of hypotension, or urinary losses. This is consistent with previous reports that ANG II is essential for the drinking response to hypotension. Furthermore, it demonstrates that ANG II is not merely permissive but probably the signal controlling water intake when arterial pressure is reduced below normal.

Pressor action of intravenous angiotensin II reduces drinking response in rats.

We investigated whether the pressor response to intravenous angiotensin II (ANG II) suppresses drinking. All experiments were done on conscious water-replete rats (200-400 g) with chronic vascular cannulas. Two rates of ANG II infusion (16.7 and 100 ng/min for 90 min) were tested; captopril (0.33 mg/min) was infused simultaneously to prevent endogenous production of ANG II. Both doses of ANG II increased mean arterial pressure (MAP) by 40-50 mmHg for the duration of the infusions, but water intakes were small. The drinking response was increased as much as fivefold, however, when the pressor response was reduced by injecting either isoproterenol (0.01 or 0.1 mg/kg, sc), diazoxide (20, 30, or 75 mg/kg, sc), or minoxidil (10 mg/kg, ip) 15 min after starting the ANG II infusion. The closer MAP was returned to normal, the greater was the drinking response. Since lowering MAP also reduced urinary water losses, net fluid intake increased even more dramatically. It is unlikely that the vasodilators directly stimulated thirst in the experiments because the dose of captopril used completely blocked drinking to these agents given alone. A situation of high circulating levels of ANG II but with MAP near or below normal more closely resembles physiological conditions of dehydration. Our results demonstrate that intravenous ANG II is a very potent dipsogen under these conditions.

The short-term effect of captopril on salt and water intake in the rat is not taste-specific.

We have investigated the extent to which captopril's short-term (1 h) effects on salt and water intake in the rat are caused by effects on taste. In single-bottle tests a low dose of captopril (0.5 mg/kg s.c.), which blocks the synthesis of angiotensin II in the blood but not the brain, increased equally the intakes of water, 0.05, 0.15, 0.30 and 0.45 M NaCl, 0.3 M KCl, 10 mM HCl, 0.14 mM quinine hydrochloride and 0.1 mM saccharin solutions without changing the animals' preference for or aversion to each with respect to water. In two-choice tests this dose increased water but not 0.15 or 0.45 M NaCl intake. A large dose of captopril (100 mg/kg s.c.), to block the synthesis of angiotensin II also in the brain, did not enhance water or NaCl intake. Neither dose affected NaCl or water intake by rats drinking in response to 2 M NaCl, 5 ml/kg i.p. We conclude that during the first hour following injection captopril has no major effect on taste perception or preference in the rat and does not stimulate sodium appetite in the sodium-replete rat. Our results support the hypothesis that low doses of captopril increase fluid intake by enhancing the synthesis of angiotensin II in the brain.

Increased or decreased thirst caused by inhibition of angiotensin-converting enzyme in the rat.

We have investigated the effects on water intake of subcutaneous (S.C.) injections of low (0.5 mg/kg) and high (100 mg/kg) doses of captopril, an inhibitor of angiotensin-converting enzyme (CE). Low doses block the synthesis of angiotensin II only in the circulation whereas high doses block CE in both the blood and the brain. The low dose of captopril enhanced drinking in response to three hypotensive drugs, isoprenaline (0.1 mg/kg, S.C.), phentolamine (5 mg/kg, S.C.) and serotonin (2 mg/kg, S.C.), whereas the high dose of captopril abolished drinking in response to these stimuli. The low dose of captopril also enhanced drinking in response to histamine (0.25-5.0 mg/kg, intraperitoneal, I.P.), but in this case the high dose of captopril only partially reduced the drinking response. The low dose of captopril enhanced drinking after 24 h water deprivation but high doses had no significant effect on deprivation-induced thirst. Hypovolaemia was produced either by injecting polyethylene glycol (30% w/v, 10 ml/kg) S.C. or by replenishing the cellular deficit in water-deprived rats with 10 ml water (by gavage). The low dose of captopril enhanced the drinking response to hypovolaemia but the high dose had no significant effect. Neither the high nor the low dose of captopril significantly affected drinking in response to cellular dehydration caused by injecting 2 M-NaCl (2 ml) I.P. or by replenishing the extracellular deficit in water-deprived rats (10 ml balanced salt solution by gavage). Nephrectomy (but not ligation of the ureters) or injections of propranolol (5 mg/kg, S.C.) to prevent renin secretion prevented the enhancement of deprivation-or serotonin-induced thirst by the low dose of captopril. The low dose of captopril did not enhance drinking in response to I.V. injections of renin (1 Goldblatt unit), or intracerebroventricular (I.C.V.) injections of angiotensin I or II. The high dose of captopril blocked drinking in response to I.V. injections of renin or I.C.V. injections of angiotensin I but did not reduce drinking in response to angiotensin II, I.C.V. These results are consistent with the hypothesis that blocking CE only in the circulation enhances drinking in response to hypotension or hypovolaemia because angiotensin I, accumulating in high concentration in the blood, enters the brain and is converted intracerebrally to angiotensin II. These findings suggest that the enhancement of drinking caused by low doses of captopril s.c. is a sensitive indicator of whether the renin- angiotensin system participates at all in the regulatory response to a particular stimulus to drink.(ABSTRACT TRUNCATED AT 400 WORDS)

Effects of systemic and intracranial inhibition of angiotensin-converting enzyme on isoproterenol-induced drinking in the rat.

We have investigated the effects of separate and combined s.c. and intracerebroventricular (i.c.v.) injections of captopril, an inhibitor of angiotensin I-converting enzyme, on isoproterenol-induced thirst. Whereas s.c. injections of captopril (0.5 mg/kg) increased drinking, combined s.c. and i.c.v. (20 micrograms) injections of captopril nearly abolished drinking to isoproterenol (0.1 mg/kg s.c.). This inhibition was not caused by general debility of the rats since the same treatment did not reduce drinking to 12h water deprivation. Intracerebroventricular injection of 20 micrograms captopril alone also greatly reduced isoproterenol-induced drinking, perhaps because it leaked into the circulation; captopril i.c.v. also reduced the pressor response to i.v. injection of hog renin (0.1 Goldblatt Unit) by about 65%. These results support the hypotheses that the renin-angiotensin system participates in the stimulation of drinking by isoproterenol and that the enhancement of drinking caused by inhibition of CE only in the circulation is the result of increased synthesis of angiotensin II in the brain.

Effects of captopril on salt appetite in sodium-replete rats and rats treated with desoxycorticosterone acetate (DOCA).

Captopril (30 mg/kg daily by gavage), an orally active inhibitor of angiotensin-converting enzyme, reduced the intake of 0.15 M NaCl in rats treated with desoxycorticosterone acetate (2.5 mg/day S.C.). However, this is probably not a specific effect on salt appetite as captopril (30 or 60 mg/kg daily) did not reduce the intake of less palatable 0.5 M NaCl in desoxycorticosterone acetate-treated rats (1 or 2.5 mg daily). In contrast, captopril given alone (30 or 60 mg/kg daily) consistently caused a 3- to 5-fold increase in intake of 0.5 M NaCl that usually began on the 1st day and persisted until treatment stopped (1-3 weeks). Sodium intake and sodium excretion increased concomitantly, but the stimulation of salt appetite occurred even in the absence of sodium depletion. Also, the effect on salt appetite was specific; captopril did not increase the intake of 0.5 M KCl or 0.03 M sucrose. This potent and specific stimulation of sodium intake by oral treatment with an inhibitor of the renin-angiotensin system may be caused by a paradoxical increase in the synthesis of angiotensin II in the brain.

Neuropeptides and thirst.

A number of neuropeptides have been found to affect fluid intake when injected directly into the brain of various vertebrate species. These include: angiotensin II and its peptide precursors; the tachykinins Substance P, eledoisin and physalaemin; the opioid peptides met- and leu-enkephalin and beta-endorphin; bombesin; neurotensin; and vasopressin. Some of these stimulate drinking, some inhibit water intake, and the tachykinins have opposite effects on thirst depending on the species tested. Very little is known about the site or mechamism of action of most of these peptides or if their effects on thirst are physiological. The exception is angiotensin II, a peptide hormone that is synthesized in the blood in response to hypovalaemia or hypotension and is involved in many aspects of the regulation of blood volume and pressure. Angiotensin II injected intravenously or intracranially stimulates drinking in all reptiles, birds and mammals tested. In addition to its role as a hormone, angiotensin II may also function as a neurotransmitter or neuromodulator, since all of the enzymes and precursors necessary for its synthesis have been found in the central nervous system.