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1. A dietary combination of high salt and low potassium (HSLK) exacerbates hypertension in Dahl salt-sensitive (DS) rats and renders previously normotensive Dahl salt-resistant (DR) rats hypertensive. In both strains, the severity of hypertension correlates with urinary calcium loss. However, the magnitude of excretory calcium losses is significantly greater in DS rats and is potentiated by chemical sympathectomy in both strains. 2. We hypothesized that a defect in vitamin D metabolism may underlie the observed strain-dependent differences in calcium balance. 3. Arterial blood pressure (ABP), water and mineral balance and serum concentrations of 1,25-dihydroxyvitamin D3 (1,25(OH)2 D3) and 25-hydroxyvitamin D3 (25(OH)D3) were measured in intact and chemically sympathectomized (6-hydroxydopamine; 6-OHDA) DS and DR rats after 8 weeks on a HSLK diet. 4. Chronic ingestion of this diet resulted in marked and moderate levels of hypertension in DS and DR rats, respectively. The hypertension was abated and eliminated by 6-OHDA in the DS and DR strains, respectively. Independent of treatment, DS rats had significantly higher urinary excretion of calcium and reduced intestinal absorption of the ion compared with DR rats. The DS rats had significantly higher serum levels of 1,25(OH)2 D3 and markedly lower serum levels of 25(OH)D3 than DR rats. Chemical sympathectomy tended to increase 1,25(OH)2 D3 and to decrease 25(OH)D3 levels in both strains. 5. These data indicate a genetic difference in vitamin D metabolism between DS and DR rats. The abnormally elevated levels of 1,25(OH)2 D3 in DS rats may be an appropriate compensatory response to excessive excretory calcium loss and reduced target organ sensitivity to the hormone and may, maladaptively, directly contribute to hypertension, by stimulating vascular smooth muscle contractility.

Objective: Atrial natriuretic peptide (ANP) lowers arterial blood pressure (ABP) chronically, in association with vasodilation of the resistance vasculature. The mechanism mediating the chronic relaxant effect of ANP is likely indirectly mediated by interactions with tonic vasoeffector mechanisms, inasmuch as the resistance vasculature is relatively insensitive to direct cGMP-mediated relaxation by ANP. On the basis of evidence that ANP has widespread sympatholytic activity, the current study investigated whether the chronic hypotensive effect of ANP is mediated by attenuation of tonic cardiovascular sympathetic tone. Methods: Total plasma catecholamine concentration and changes in basal ABP and heart rate (HR) following autonomic ganglionic blockade were measured as indices of underlying sympathetic nerve activity in hypotensive ANP-overexpressing transgenic mice (TTR–ANP), hypertensive ANP knockout mice (−/−) and the genetically-matched wild type (NT and +/+, respectively) control mice. Pressor and chronotropic responses to norepinephrine infusion were measured in ganglion-blocked mice of all genotypes, and norepinephrine receptor binding was assessed in representative tissues of −/− and +/+ mice, in order to determine whether peripheral adrenergic receptor responsiveness is altered by ANP-genotype. Results: Basal ABP was significantly lower in TTR–ANP and higher in −/− compared to their wild-type controls. Basal HR did not differ significantly between mutant and control mice. Autonomic ganglionic blockade reduced ABP and HR in all genotypes, however, the relative decrease in ABP was significantly smaller in TTR–ANP and greater in −/− mice than in their respective controls. Total plasma catecholamine was significantly higher in −/− than in +/+ mice but did not differ significantly between TTR–ANP and NT mice. Norepinephrine infusion during ganglionic blockade elicited quantitatively similar pressor and chronotropic responses in mutant and control mice. Tissue norepinephrine binding did not differ significantly between −/− and +/+ mice. Conclusions: The present study shows that differences in endogenous ANP activity in mice, resulting in chronic alterations in ABP are accompanied by directional changes in underlying cardiovascular sympathetic tone, and suggests that the chronic vasodilator effect of ANP is, at least partially, dependent on attenuation of vascular sympathetic tone, possibly at a prejunctional site(s).

Atrial natriuretic peptide (ANP) is thought to play a role in renal regulation of salt balance by reducing tubular reabsorption of sodium and chloride. Therefore, in the chronic absence of this hormone, a defect of salt excretion should be evident. We used an ANP gene deletion model to test this premise. F2 homozygous mutant mice (-/-) and their wild-type littermates (+/+) were fed an 8% NaCl diet prior to an acute infusion of isotonic saline. Arterial blood pressures, renal excretions of salt and water, as well as collecting duct transport of fluid and electrolytes were measured. Pressures were significantly higher in -/- compared with +/+ mice (139 ± 4 vs. 101 ± 2 mmHg; 1 mmHg = 133.3 Pa). There was no difference in glomerular filtration rate (-/- = 0.84 ± 0.06; +/+ = 0.81 ± 0.04 mL·min-1·g-1 kidney weight). In the collecting duct, sodium and chloride reabsorptions were significantly higher in the -/- group than in the +/+ group. As a result, natriuresis and chloruresis were relatively reduced (UNaV: -/- = 8.6 ± 1.1; +/+ = 14.0 ± 1.1; UClV: -/- = 10.1 ± 1.4; +/+ = 16.0 ± 1.1 µmol·min-1·g-1 kidney weight). We conclude that the absence of endogenous ANP activity in mice on a high-salt diet subjected to acute saline infusion causes inappropriately high reabsorption of sodium and chloride in the medullary collecting duct, resulting in a relative defect in renal excretory capacity for salt.Key words: high-salt diet; water, sodium, chloride, and potassium transport.

Atrial natriuretic peptide (ANP) exerts a chronic hypotensive effect due to a decrease in total peripheral resistance (TPR). This study examines if chronic ANP-dependent vasodilation is attributable to differences in the cardiovascular regulatory activity of vascular endothelium (VE), based on evidence that ANP affects synthesis/release and target cardiovascular effects of endothelin-1 (ET-1), C-type natriuretic peptide (CNP), and nitric oxide (NO). To determine if the synthetic activity of resistance vasculature VE is chronically altered by plasma ANP activity, we measured ET-1, CNP, and endothelial constitutive NO synthase (ecNOS) concentration and total NOS enzyme activity in homogenates of kidney, heart, lung, hindquarter skeletal muscle, and brain from hypotensive transgenic mice with elevated plasma ANP, hypertensive knockout mice (−/−) characterized by the absence of ANP, and the corresponding normotensive wild-type (NT, +/+) mice. Tissue distribution and abundance patterns of ET-1, CNP, ecNOS, and NOS enzyme activity were comparable between the different genotypes and did not differ significantly between mutant and control mice. Antagonism of ETA/Breceptors in −/− and +/+ mice in vivo with SB-209670 reduced arterial blood pressure (ABP) significantly and comparably in both genotypes (−27 ± 4 and −25 ± 2% change for −/− and +/+ mice, respectively) independent of any significant changes in heart rate (HR) (−6 ± 8 and −4 ± 4% change for −/− and +/+ mice, respectively). Immunoneutralization of CNP-specific guanylate cyclase-linked receptors (GC-B) with monoclonal antibodies (3G12) increased ABP slightly, but not significantly, by similar relative amounts in both −/− (10 ± 6% change) and +/+ mice (8 ± 3% change), without changing HR significantly (4 ± 1% change for both +/+ and −/− mice). Inhibition of NOS activity (by NG-nitro-l-arginine methyl ester) significantly increased ABP, but the changes were comparable between −/− (53 ± 5% change) and +/+ mice (50 ± 6% change) and occurred in the absence of significant changes in HR (−1 ± 5 and 7 ± 5% change for −/− and +/+ mice, respectively). We conclude that the differences in ABP associated with chronic variations in endogenous ANP activity are not due to alterations in synthesis or responsiveness of the cardiovascular system to the effects of ET-1, CNP, or NO.

Atrial natriuretic peptide (ANP), a peptide hormone produced by the heart, exerts a chronic hypotensive effect. Knockout mice with a homozygous disruption of the pro-ANP gene (−/−) are incapable of producing ANP and are hypertensive relative to their wild-type (+/+) siblings. Previous studies showed that arterial blood pressure (ABP) was further increased in conscious −/− mice kept for 2 wk on 2% salt, but not in anesthetized −/− mice after 1 wk on 8% salt. To determine whether inconsistencies in observed effects of salt on ABP of −/− mice are due to duration of increased salt intake and/or the state of consciousness of the animals, we measured ABP from an exteriorized carotid catheter during and after recovery from anesthesia with ketamine-xylazine in adult +/+ and −/− mice kept on low (LS; 0.008% NaCl)- or high (HS; 8% NaCl)-salt diets for 3–4 wk. Conscious ABP ± SE (mmHg) of +/+ mice did not differ significantly on either diet (HS, 113 ± 3; LS, 110 ± 5). However, on HS diet −/− mice had significantly higher ABP (135 ± 3; P < 0.001) than both −/− (115 ± 2) and +/+ (110 ± 5) mice on LS diet. Anesthesia decreased ABP in all groups, but the genotype- and diet-related differences were preserved. Plasma renin activity (PRA, ng ANG I ⋅ ml−1 ⋅ h−1) in blood collected at termination of experiment was appropriately different on the 2 diets in +/+ mice (HS, 4.9 ± 1.9; LS, 21 ± 2.8). However, PRA failed to decrease in −/− mice on HS diet (HS, 18 ± 2.9; LS, 19 ± 3.7). Independent of genotype, concentration of endothelin-1 (ET-1, pg/mg protein) and endothelial constitutive NOS (ecNOS, density/100 μg protein) was significantly elevated in kidneys of mice fed on HS diet (ET-1 −/−, 31 ± 4.7 and +/+, 32 ± 4.1; ecNOS −/−, 160 ± 19 and +/+, 156 ± 19) compared with mice fed on LS diet (ET-1 −/−, 19 ± 1.9 and +/+, 21 ± 1.8; ecNOS −/−, 109 ± 13 and +/+, 112 ± 18). We conclude that, regardless of the state of alertness, −/− mice develop salt-sensitive hypertension after prolonged feeding on HS, in part due to their inability to reduce PRA, whereas the specific renal upregulation of ecNOS and ET-1 in response to HS intake may be an ANP-independent adaptive adjustment aimed at improving kidney function and counteracting the pressor effect of salt.

Release of atrial natriuretic factor (ANF) from the heart is primarily affected by myocyte stretch. In addition, ANF release can be modulated by a variety of hormones and neurotransmitters, but the mechanisms involved in such modulation are not completely understood. In the present study, we investigated the effect of inhibition of cyclooxygenase activity on release of ANF from the isolated, spontaneously beating rat heart: (1) during basal conditions; and (2) in response to arginine vasopressin (AVP), acetylcholine (ACh) and angiotensin II (ANG II), in order to determine if cardiac prostaglandin synthesis is involved in modulation of basal and hormone-mediated ANF secretion. Basal secretion in the time controls remained stable for the duration of the experiment. AVP, ACh and ANG II reduced basal secretion significantly by 58 ± 4%, 51 ± 6% and 26 ± 8%, respectively, independently of concomitant changes in coronary flow and heart rate. Inhibition of cyclooxygenase with indomethacin (1 · 10−5 M) decreased basal ANF release by 38 ± 6%, indicating that basal secretion requires prostaglandin production. The effects of AVP, ACh and ANG II were maintained during perfusion with indomethacin, suggesting a common mechanism of action which operates via inhibition of cyclooxygenase. Based on our previous findings that the effects of indomethacin, AVP and ACh are overcome by inhibition of NO/EDRF synthesis, we suggest a common mechanism of action by means of which NO/EDRF mediates the effects of these agents by inhibiting cyclooxygenase activity.

Transgenic mice overexpressing an atrial natriuretic factor (ANF) fusion gene (TTR-ANF) and their nontransgenic siblings were placed on a high- (8%) or low (< 0.008%)-salt diet for 14 days to determine whether the lifelong elevation of ANF in the transgenic animals compromised their ability to maintain fluid-electrolyte balance. Steady-state dietary intake and urinary output of sodium and chloride were not statistically different between TTR-ANF and control groups on either diet. By contrast, water intake and urine volume were markedly elevated in the TTR-ANF group on either diet. Arterial blood pressure, measured in anesthetized mice at the end of the dietary regimen, was significantly and similarly reduced in the TTR-ANF compared with control groups on either diet, although high salt intake was associated with increased pressure in both groups. Renal excretion of fluid and electrolytes was studied in the anesthetized mice before and after acute blood volume expansion. Although the absolute increase in fluid and electrolyte excretion was much greater on the high- than on the low-salt diet in both groups, TTR-ANF mice had an exaggerated response relative to controls on either diet. On the basis of these results, we conclude the following. 1) When they are stimulated, renal salt-conserving mechanisms are sufficiently powerful to overcome the expected natriuretic effects of chronic elevation of plasma ANF; however, the natriuretic potential of ANF can be expressed in the short term when the counterregulatory mechanisms are inactivated. 2) ANF exerts a chronic hypotensive effect that is independent of changes in renal salt excretion.(ABSTRACT TRUNCATED AT 250 WORDS)

Weanling male inbred Dahl rats (Jr salt-sensitive (S) and salt-resistant (R) strains) were placed on high (4%, HK) and low (0.2%, LK) potassium diets for 4 weeks. Both diets contained 8% sodium chloride, 2.5% calcium, 0.8% magnesium, and 2.0% phosphorous. Balance studies were carried out during the final week on the diets. Mean arterial blood pressure was determined, and dietary intake and urinary output of water, sodium, chloride, potassium, calcium, magnesium, and phosphate were monitored daily during this period. The data show that blood pressures of S rats were significantly higher than those of R rats on both HK and LK diets; however, reduced dietary potassium was associated with increased blood pressure in both strains. Urinary excretions of calcium and magnesium were higher, and urinary phosphate excretion was lower, in S compared to R rats. Decreased potassium intake was associated with increased excretion of calcium, magnesium and phosphate in both strains. The changes in calcium and magnesium excretion were significantly correlated to blood pressure across strains and diets. We conclude that the effects of a high salt diet on increasing blood pressure can be potentiated by lack of potassium, even in previously salt-resistant rats. Increased blood pressure is associated with increased divalent cation excretion. It is not yet known whether this is a cause-and-effect relationship.

Abnormalities in renal handling of calcium, magnesium, or phosphate have been implicated in the development and/or maintenance of human hypertension. We have shown recently that renal excretion of these ions is correlated to blood pressure in Dahl salt-sensitive as well as salt-resistant rats. The present study was designed to determine whether renal perfusion pressure per se could affect excretion of these ions. Urinary excretion of calcium, magnesium, and phosphate was studied in anaesthetized Sprague-Dawley rats under basal conditions and during an intravenous infusion of angiotensin II (ANG II), vasopressin (AVP) or phenylephrine (PE). A cuff, placed around the aorta between the two renal arteries, allowed maintenance of normal perfusion pressure in the left kidney, while that in the right kidney was allowed to rise. Infusion of pressor agents raised mean arterial blood pressure to comparable levels (means +/- SE): ANG II (n = 7), before = 102 +/- 4, during = 133 +/- 3 mmHg, AVP (n = 8), before = 110 +/- 7, during = 136 +/- 5 mmHg, PE (n = 6), before = 111 +/- 6, during = 141 +/- 6 mmHg. Although there was no difference in excretion of calcium, magnesium and phosphate between the two kidneys under basal conditions, infusion of ANG II or PE induced hypercalciuria, hypermagnesiuria and hyperphosphaturia in the right kidney which was exposed to the increased arterial pressure. Such effects did not appear in the pressure-controlled left kidney. Infusion of AVP was associated with reduced excretion of calcium and magnesium, and increased excretion of phosphate, in the normotensive kidney. The response to the similarly increased renal perfusion pressure in this group was also reduced for calcium and magnesium, and enhanced for phosphate. The results indicate (1) renal excretion of calcium, magnesium and phosphate is renal perfusion pressure-dependent; the higher the renal perfusion pressure, the greater the excretion of these ions. (2) Independently of perfusion pressure, AVP can inhibit phosphate reabsorption and stimulate divalent cation reabsorption.

The effects of inhibition of bradykinin and prostaglandin on the renal and blood pressure responses to atrial natriuretic factor (ANF) were studied in anesthetized rats. Intraarterial infusion of the receptor antagonist D-Arg[Hyp3,Thi5,8,D-Phe7]bradykinin (BKA) at 14 μg/min, a rate sufficient to block the hypotensive response to 250 μg of bradykinin, did not affect the natriuresis due to injection of ANF (UNaV: control, before ANF, 393 ± 101, after ANF, 2322 ± 400 nmol/min; BKA, before ANF, 261 ± 72, after ANF, 2259 ± 390 nmol/min). In contrast, infusion of indomethacin (Indo) reduced the level of sodium excretion both before and especially after ANF administration (UNaV: Indo, before ANF, 75 ± 15, after ANF, 320 nmol/min). The effect of combining BKA with Indo was not different from the effect of Indo alone (UNaV: BKA + Indo, before ANF, 119 ± 26, after ANF, 469 ± 167 nmol/min). The bradykinin antagonist, with or without Indo, was associated with significant hypotension relative to control. Indo, both in the absence and presence of the antagonist, was associated with a progressive decrease in blood pressure compared with control. However, in each case the hypotensive responses to ANF were not different from those in the control group. We conclude that under the present experimental conditions bradykinin does not modify ANF-induced natriuresis. However, inhibition of prostaglandin synthesis by Indo is associated with renal salt retention, reducing natriuresis both before and after ANF administration.Key words: prostaglandin inhibition, D-Arg[Hyp3,Thi5,8,D-Phe7]bradykinin, atrial natriuretic factor.

A transgenic mouse model in which atrial natriuretic factor (ANF) expression is targeted to the liver was used to study intrarenal adjustments to the chronically elevated hormone level. Such animals, designated TTR-ANF, are characterized by reduced arterial blood pressure but similar sodium excretion compared with nontransgenic siblings. Proximal tubular micro-puncture gave the following results: single-nephron filtration rate = 12.7 ± 1.1 vs. 15.6 ± 1.9 nL/min (TTR-ANF versus nontransgenic, ns); end-proximal tubular fluid/plasma concentration ratio of inulin = 1.93 ± 0.09 vs. 1.97 ± 0.15 (ns); fractional reabsorption of sodium = 45.5 ± 2.8 vs. 46.0 ± 3.8% (ns); fractional reabsorption of chloride = 33.6 ± 3.3 vs. 32.4 ± 4.1% (ns). These data indicate that life-long elevation of plasma ANF concentration was not associated with significant alteration in single-nephron filtration rate and proximal tubular function. We conclude that compensatory anti-natriuretic mechanisms, localized downstream from the proximal tubule, can prevent ANF natriuresis.Key words: micropuncture, single-nephron filtration rate, sodium chloride reabsorption.

Objective A dietary combination of high salt and low potassium (HS-LK) exacerbates hypertension in Dahl salt-sensitive (DS) rats and renders Dahl salt-resistant (DR) rats hypertensive. In both strains, the hypertension is accompanied by remodelling of the renal resistance vasculature, and is attenuated by peripheral chemical sympathectomy. In the current study, we sought to determine whether the sympathetic nervous system is causally involved in mediating the renal vascular and haemodynamic alterations associated with HS-LK feeding in Dahl rats. Design Two groups each of DS and DR rats were maintained on HS-LK diet (8% NaCl, 0.2% KCl) for 8 weeks. One group of DS (n = 9) and DR (n = 8) were treated with 6-hydroxydopamine (6-OHDA) in 0.001 N HCl vehicle to chemically ablate peripheral sympathetic nerve terminals. The two remaining groups (n = 8 each) received equivalent injections of vehicle. Methods At the end of the dietary regimen, arterial blood pressure (ABP), glomerular filtration rate (GFR) and renal blood flow (RBF) were measured, and the structure of intra-renal resistance vessels was examined by planar morphometric analysis of coronal sections prepared from perfusion-fixed kidneys. Results Both 6-OHDA-treated and untreated DS rats presented a greater degree of intra-renal vessel remodelling characterized by reduced lumen diameter in the absence (eutrophic) or presence (hypertrophic) of cross-sectional area expansion, higher renal vascular resistance (RVR) and lower GFR and RBF than DR rats. Chemical sympathectomy increased lumen diameters and reduced vascular wall expansion, resulting in a decrease in RVR and a concomitant increase in RBF and GFR in both strains; however, the effect was more prominent in the DS rats. Conclusions We conclude that HS-LK-induced changes in intra-renal vessel structure and renal haemodynamic function in Dahl rats are, at least in part, dependent on the activity of the sympathetic nervous system.

Atrial Natriuretic Peptide (ANP) exerts a chronic hypotensive effect which is mediated by a reduction in total peripheral resistance (TPR). Mice with a homozygous disruption of the pro-ANP gene (-/-) fail to synthesize ANP and develop chronic hypertension in comparison to their normotensive wild-type (+/+) siblings. In order to determine whether alterations in basal hemodynamics underlie the hypertension associated with lack of endogenous ANP activity, we used anesthetized mice to measure arterial blood pressure (ABP) and heart rate (HR), as well as cardiac output (CO) by thermodilution technique. -/- (n = 7) and +/+ (n = 10) mice of comparable weight and age were used. Stroke volume (SV) and TPR were derived from CO, HR, and ABP by a standard formula. ABP (mm Hg) was significantly higher in -/- (132+/-4) (P0.0001) than in +/+ mice (95+/-2). CO (ml min(-1)), HR(beats min(-1))and SV (microl beat(-1)) did not differ significantly between -/- and +/+ mice (CO -/- = 7.3+/-0.5, +/+ = 8.3+/-0.6; HR -/- = 407+/-22, +/+ = 462+/-21; SV -/- = 17.6+/-1.1, +/+ = 17.6+/-1.7). However, TPR (mm Hg ml(-1) min(-1)) was significantly elevated in -/- mice (18.4+/-0.7) compared to +/+ mice (12.3+/-1) (P = 0.0003). Autonomic ganglion blockade with a mixture of hexamethonium and pentolinium was followed by comparable percent reductions in CO (-/- = 28+/-4, +/+ = 29+/-3), HR (-/- = 9+/-4, +/+ = 16+/-4) and SV(-/- = 21+/-4, +/+ = 15+/-6) in both genotypes. However, the concomitant decrease in ABP (%) in -/- (41+/-2) was significantly greater than in +/+ (23+/-4) mice (P = 0.0009) and was accompanied by a significant reduction in TPR. We conclude that the hypertension associated with lack of endogenous ANP is due to elevated TPR, which is determined by an increase in cardiovascular autonomic tone.

Dietary combination of high salt with low potassium (HSLK) exacerbates hypertension development in Dahl salt-sensitive (S) rats, and produces a mild degree of hypertension in otherwise salt-resistant (R) rats. Increased blood pressure in both strains is associated with increased urinary excretion of calcium and magnesium. The objective of this study was to determine the effect of blood pressure on body balance of these ions in Dahl rats on HSLK diet. Two groups of S and two groups of R weanlings were all placed on HSLK diet (NaCl=8%, K=0.2%) for eight weeks. One group of each strain was subjected to chemical sympathectomy with 6-hydroxydopamine (6-OHDA) to counteract hypertension development. Urinary norepinephrine was used to determine efficacy of 6-OHDA treatment. Systolic blood pressures of conscious animals were measured daily throughout the study. The last three days on the diet were used to determine total dietary intake and urinary as well as fecal excretion of sodium, calcium and magnesium. At the end of the study, extracellular fluid volume, serum aldosterone and parathyroid hormone were analyzed. Final systolic blood pressures in the 4 groups were as follows: S=235+/-9 mmHg (n=9); R=155+/-3 mmHg (n=8); 6-OHDA S=151+/-6 mm Hg (n=8); 6-OHDA R=117+/-6 mm Hg. Chemical sympathectomy decreased blood pressure in both S and R rats. There was no indication of sodium accumulation in S rats. Associated with reduced parathyroid hormone levels the S strain had significantly less positive balance for calcium than the R strain, primarily due to increased urinary excretion. A less positive balance for magnesium was also observed, due mainly to relatively reduced intestinal absorption of the ion. We conclude that the HSLK diet is associated with inappropriate activation of the sympathetic nervous system and increased arterial pressure in both strains. In addition, since divalent cations may influence blood pressure, we suggest that the observed abnormalities in calcium and magnesium metabolism might independently promote hypertension development in the S strain.

Antidiuretic hormone leads to an increase in the permeability for water and urea in the inner medullary collecting duct. Hence, urea may not be an "effective" osmole in the inner medulla during maximal renal water conservation. Accordingly, the purpose of this study was to evaluate whether differences in the rate of urea excretion would influence maximum renal water conservation in humans. In water-deprived rats, the concentration of urea and total osmolality were somewhat higher in the urine exiting the inner medullary collecting duct than in interstitial fluid obtained from the entire papillary tip. Nevertheless, the "nonurea" (total osmolality minus urea in millimolar terms) osmolality was virtually identical in both locations. Chronically fasted human subjects that were water-deprived for 16 h had a lower rate of urea excretion (71 +/- 7 versus 225 +/- 14 mumol/min) and a somewhat lower urine osmolality (745 +/- 53 versus 918 +/- 20 mosmol/kg H2O). Nevertheless, they had identical urine flow rates (0.5 +/- 0.01 and 0.5 +/- 0.02 ml/min, respectively), and their nonurea osmolality also was similar (587 +/- 25 and 475 +/- 14 mosmol/kg H2O, respectively) to the water-deprived normal subjects. The composition of their urine differed in that the principal nonurea osmoles became NH4+ and beta-hydroxybutyrate rather than Na and C1. During water deprivation in normal subjects, the ingestion of urea caused a twofold rise in urine flow rate, a fall in the nonurea osmolality, and a rise in the rate of excretion of nonurea osmoles. The nonurea osmolality of the urine, and presumably the medullary interstitial fluid as well, was inversely related to the urea excretion rate. In chronic fasting, the nature, but not the quantity, of nonurea osmoles changed. The similar minimum urine volume was predictable from an analysis based on nonurea osmole considerations.

During dietary salt deprivation, the sympathetic nervous system and the angiotensin-aldosterone system are stimulated. Both systems are thought to be essential for maximal salt conservation by the kidney. To study their relative contributions, we produced negative salt balance in rats by intraperitoneal injection of furosemide, followed by a low-salt diet (

Chronic salt depletion was used as a model to study the mechanism of renal resistance to the natriuretic effect of atrial natriuretic factor (ANF). Rats were pretreated with furosemide and placed on a low-salt diet (

Inbred Dahl salt-sensitive (S) and sal-resistant (R) rats were fed with either high (4% K; HK) or low (0.2% K; LK) potassium diets (both contained 8% salt, 2.5% calcium, 0.8% magnesium, and 2.0% PO4) for 4 wk. During the last week, systolic blood pressure was measured in conscious animals. External diameter of renal vessels (ED) was used to divide renal vessels into four subgroups: less than 25 microns; greater than 25 microns but less than 55 microns; greater than 55 microns but less than 150 microns; and greater than 150 microns. Luminal diameter (LD), cross-sectional wall area (WAC) and wall-tolumen ratio (W/L) were measured or calculated. Glomerular volume (VG) was measured. Data showed that blood pressures of S rats were significantly higher than those of R rats on both diets. Reduced dietary potassium was associated with increased blood pressure in both strains. Changes in renal vessels were characterized by bigger lumen and greater wall area in S rats, compared with R rats, especially in the smallest vessels. Low potassium intake was associated with reduced lumen diameter and enlarged W/L ratio with or without changing WAC in both strains. VG showed no significant differences between strains or between diets. It was concluded that in addition to the genetically determined differences in renal vascular structure between S and R rats, dietary potassium depletion may engender further renal injury, exacerbating hypertension.

1. Chronic reduction of salt intake can reduce the natriuretic effect of exogenously administered atrial natriuretic factor. The purpose of this study was to elucidate the intrarenal site(s) of such atrial natriuretic factor resistance. Renal clearance and collecting duct microcatheterization experiments were made before and during infusion of atrial natriuretic factor in three groups of rats: group 1 consisted of rats fed a high salt diet (8% NaCl) for 1 week before the experiment; group II were fed a low salt diet (< 0.008%); group III received the same low salt diet, but were acutely replenished with salt at the time of experiment. 2. Baseline sodium chloride excretion was 6480 ± 810 nmol min−1g−1 kidney weight in group 1 compared to 99 ± 16 in group 1. Fractional re-absorptions in the medullary collecting duct were 37 ± 6% and 95 ± 2% of delivered load, respectively (P < 0.05). The fractions of filtered sodium remaining at the beginning of the medullary duct were 6.6 ± 1.0% of filtered load in group 1 and 2.7 ± 0.7% in group II (P < 0.05), indicating increased tubular reabsorption in group II, not only in the medullary duct, but also in upstream nephron segments. 3. During infusion of atrial natriuretic factor, marked saluresis (13240 ± 750 nmol min−1 g−1 kidney weight), together with decreased fractional reabsorption at both sites (duct, −13 ± 9%; upstream remainder, 7.9 ± 0.7%; P < 0.05 each, compared to corresponding control values) was found in group 1, whereas the excretory (150 ± 28 nmol min−1 g−1 kidney weight), and the tubular transport (duct = 84 ± 3%; upstream remainder =2.2 ± 0.4%) changes were quantitatively insignificant in group II. Glomerular filtration rate was increased in group 1 from 1.07 ± 0.03 to 1.26 ± 0.04 ml min−1g−1 kidney weight (P < 0.05), but not in group II (0.93 ± 0.07 to 0.96 ± 0.09, not significant). 4. In group III, acute salt replenishment was associated with increased excretion (1940 ± 440 nmol mm−1 g−1 kidney weight, P < 0.05 compared to group II) and with reduction of tubular reabsorption in the collecting duct only (69 ± 8%, P < 0.05). Infusion of atrial natriuretic factor in this group further increased natriuresis (7810 ± 780 nmol min−1 g−1 kidney weight) and decreased tubular reabsorption in the duct (−32 ± 22%, P < 0.05 compared to the corresponding control value). 5. We conclude that chronic salt deprivation can effectively prevent, via a rapidly reversible counter-regulatory mechanism, the expected actions of atrial natriuretic factor on sodium reabsorption in the medullary collecting duct. Operation of such a mechanism may explain salt retention despite elevated endogenous levels of atrial natriuretic factor in pathological states such as congestive heart failure and liver cirrhosis.

In addition to atrial natriuretic peptide (ANP 99–126 ) itself, linear peptide fragments from its N-terminal prohormone segment (pro-ANP) have been reported to have biological activity. In vivo, diuresis and natriuresis, as well as hypotension have been observed. In vitro, sodium uptake into medullary collecting duct cells was inhibited, and tone of vascular smooth muscle was reduced, associated with activation of guanylate cyclase. Such previous studies have used heterologous peptides and species, e.g., human pro-ANP 1–30 or pro-ANP 31–67 , tested in rat, pig, or dog. The present experiments were designed to test whether rat pro-ANP 1–30 or pro-ANP 31–67 were natriuretic and hypotensive in rats, whether the two peptides showed specific binding to plasma membranes from rat kidney cortex or aorta, and whether they affected particulate guanylate cyclase activity in rat glomerular membranes. To extend in vitro results from the literature, the effect of human pro-ANP 31–67 on transport in the rat medullary collecting duct in vivo was also tested. Although rat ANP 99–126 , as expected, increased diuresis and natriuresis, associated with inhibition of transport in the medullary collecting duct, in identically treated rats human pro-ANP 31–67 was without effect. Similarly, only the ANP 99–126 infusion resulted in reduction of arterial blood pressure. Furthermore, no diuretic, natriuretic, or hypotensive responses were observed in rats infused with either rat pro-ANP 31–67 or pro-ANP 1–30 . In plasma membranes from rat kidney cortex or aorta, neither of the rat prosegments showed specific binding, or interference with ANP 99–126 receptors. Finally, in contrast to ANP 99–126 , neither of the prosegments was able to increase basal guanylate cyclase activity in rat glomerular membranes. Therefore, under our experimental conditions we were unable to replicate the earlier results. This study thus does not support a regulatory role for pro-ANP fragments in blood volume or blood pressure homeostasis.

A selective antagonist for the cGMP-linked ANF receptor was used to assess inhibition of cardiovascular and renal actions of atrial natriuretic factor (ANF). Two groups of anesthetized rats were injected with antagonist or vehicle, respectively, prior to an infusion of ANF. A third group received neither antagonist injection nor ANF infusion and served as a time control. Compared to ANF infusion alone, prior antagonist administration was associated with significant reduction of both the hypotension and hemoconcentration following peptide infusion, although significant residual effects were still present. Glomerular filtration rates during ANF infusion were significantly lower in the antagonist group. The increases in urinary salt and water excretion were also partially blocked by the antagonist. Microcatheterization studies showed significant partial reversal of ANF-induced inhibition of sodium chloride and water reabsorption in the medullary collecting duct. We conclude that the antagonist is an effective specific blocker of the cardiovascular, renal hemodynamic, and tubular effects of ANF, providing a useful new tool to elucidate the regulatory roles of this peptide hormone system.

The renal response to iso-oncotic blood volume expansion with bovine serum albumin was studied in anaesthetized homozygous Brattleboro (DI) rats after acute (1 day) pretreatment with 1 U arginine-vasopressin (AVP) compared to heterozygous controls. In AVP-treated DI (DI + AVP) rats, basal urine flow as well as urinary sodium, chloride, and potassium excretion, were not different from controls. Diuresis and kaliuresis induced by volume expansion were blunted in DI + AVP rats. However, natriuresis and chloruresis were exaggerated in DI + AVP rats. They increased faster, reached a higher maximum, but declined earlier, compared to controls. The blunted diuresis resulted in a positive volume balance by the end of the experiment in DI + AVP rats, whereas the controls showed restoration of normal balance. Significant retention of sodium and chloride was observed in controls, but not in DI + AVP rats, over the time of the experiment. DI + AVP rats lost significantly less potassium than controls during the experiment. As judged from the lithium clearance method, the exaggerated saluresis in DI + AVP rats was mainly due to a reduced proximal sodium reabsorption. Plasma immunoreactivity of atrial natriuretic peptide (ANP) rose during blood volume expansion and fell in the recovery period. It was not different between AVP-treated DI rats and controls at any time of the experiment. Inulin clearance was slightly, but not significantly, lower in DI + AVP rats and increased after blood volume expansion in DI + AVP rats only.(ABSTRACT TRUNCATED AT 250 WORDS)

Nephron site responsible for the reduced kaliuretic response to mineralocorticoids during hypokalemia in rats. Rats with hypokalemia induced by eating a low-K diet have a diminished kaliuretic response to mineralocorticoids. The purpose of this study was to determine if this was the due to a lower rate of net secretion of K in the cortical collecting duct (CCD) and/or an enhanced rate of reabsorption of K in the medullary collecting duct (MCD). Secondary active secretion of K in the CCD raises the [K] in the lumen as compared to the plasma ((TF/P) K ). If the (TF/P) K is greater than 1, there was secondary active secretion of K in this nephron segment. The (TF/P) K in the CCD was measured by microcatheterization of the collecting duct. Three groups of rats were studied: rats on a low-K diet with and without the acute administration of DOCA, and rats on a normal-K diet treated with DOCA on a chronic basis. Rats on the low-K diet had a (TF/P) K of 0.8 ± 0.11; this value did not rise to values significantly greater than 1 after the acute administration of DOCA (1.4 ± 0.35). In contrast, chronic administration of DOCA to rats fed a normal-K diet did result in a (TF/P) K which was significantly greater than unity (3.1 ± 0.39). The degree of hypokalemia was not significantly different in these rats. The absolute and fractional reabsorption of K in the MCD was not different in the rats on the low-K diet with or without DOCA. We conclude that the nephron segment which is responsible for the reduced kaliuretic response to mineralocorticoids is the CCD.

A method is described that allows perfusion of the inner medullary collecting duct (IMCD) of the rat kidney in situ and in vivo. Fine polyethylene catheters connected to a microperfusion pump were inserted into collecting ducts via the openings at the exposed papilla tip. Perfusate contained 22Na as well as [3H]inulin. During perfusion at 30 nl/min, urine was simultaneously collected. A decrease in the Na-to-inulin concentration ratio in the urinary sample, compared with the perfusate, was taken as indicating unidirectional efflux of Na from the perfused duct system. The effects of luminal amiloride (2 X 10(-4) M) or atrial natriuretic factor (ANF, 10(-8) M) were studied. Compared with control perfusions, both agonists reduced Na efflux from the IMCD to approximately 50%, indicating luminal sites of action. Combination of amiloride and ANF at their respective concentrations had no further effect. The lack of statistically significant additivity suggests, but does not prove, that ANF, administered from the luminal side, is able to block amiloride-sensitive Na channels in the apical membrane of IMCD cells.

The role of the medullary collecting duct in pressure natriuresis has not been established. In vivo microcatheterization was used to study the effect of an acute increase in blood pressure induced by bilateral carotid artery and vagal nerve ligation on medullary collecting duct function in anaesthetized rats. Increased fluid and electrolyte excretion during pressure natriuresis were accompanied by increased delivery of water, sodium, chloride, and potassium to the beginning of the medullary collecting duct, a change that was significantly greater than in a second series of time-control animals. These increases in delivery were within the range for which constant fractional NaCl reabsorption had been found previously. However, during increased perfusion pressure, reabsorption of both sodium and chloride in the medullary collecting duct as a fraction of delivered load were reduced from 81 ± 4.1 to 51 ± 9.3% (p

Renal and hemodynamic effects of an intravenous infusion of atrial natriuretic factor (ANF) (8 micrograms/h) were studied in homozygous Brattleboro rats, which lack endogenous vasopressin. Heterozygous rats were used as controls. ANF-induced increases in sodium, chloride and volume excretion were higher, whereas changes in potassium excretion were lower in homozygous, as compared to heterozygous rats. The initial decrease in arterial blood pressure after ANF infusion was greater in the homozygous group, whereas there were no differential effects on heart rate. Inulin clearance, as well as clearance and fractional excretion of lithium were not significantly different between groups. The results indicate that Brattleboro rats show an exaggerated diuretic as well as saluretic response to ANF. They suggest that these effects are localized in the distal nephron and may be due to the known anatomical abnormalities in juxtamedullary nephrons of Brattleboro rats.

Adrenalectomized rats, kept on tap water for 3 days and infused with the glucocorticoid dexamethasone during the experiment, were compared to similarly treated sham-operated rats. Using the microcatheterization technique, reabsorption of fluid, sodium and potassium in the medullary collecting duct was studied before and after infusion of donor blood (2.3% of body weight). Before intravascular volume expansion sodium excretion in adrenalectomized rats was greater than in sham-operated ones. Extensive overlap between the two groups made this difference not statistically significant. However, the fraction of filtered sodium excreted was significantly greater after adrenalectomy, indicating the expected tubular transport defect. Fluid reabsorption from the medullary collecting-duct system was comparable in both series. Adrenalectomy did not inhibit net sodium reabsorption from the inner medullary duct, although reduction of Na transport in the outer medullary collecting system could be inferred. Renal excretion of potassium was not associated with net K secretion in the collecting duct in either group. During hypervolemia induced by intravenous infusion of donor blood, marked diuresis, natriuresis and kaliuresis were observed in all animals, associated with inhibition of net fluid and sodium reabsorption along the collecting system in both inner and outer medulla. Small, but statistically significant secretion of potassium became evident. The relatively reduced renal response in adrenalectomized animals could be attributed in part to a decreased filtered load compared to sham-operated rats. It is concluded: (1) that lack of mineralocorticoid does not prevent the normal fluid and sodium reabsorption from the lumen of the inner medullary collecting system, and (2) that the inhibition of this reabsorption consequent to hypervolemia is independent of changes in plasma aldosterone levels.

The natriuretic activity shown in earlier cross-circulation experiments to develop in the blood of rats undergoing sustained vascular expansion has been further characterized. Following whole blood infusion and urine reinfusion of the donor rat, a cross-circulated isovolaemic partner exhibits a natriuresis of gradual onset, requiring 60–80 min to reach a peak (0.04–3.24 μequiv./min per gram kidney weight). The gradual feature of recipient natriuresis was unchanged by infusing the donor rat 1 h prior to cross-circulation. Once developed, the recipient natriuresis was sustained when fluid depletion by the renal response was prevented, but was reversible with a half-life of about 30 min on interruption of the cross-circulation; reconnection of cross-circulation promptly restored recipient natriuresis. No significant natriuresis (0.04–0.16 μequiv./min per gram kidney weight) occurred in the recipients if the donors in comparable cross-circulations were not infused. These findings confirm that a natriuretic activity develops in the blood as a consequence of vascular expansion, and reveal that the activity has a slowly developing and reversible action on the renal natriuretic mechanism.

Microcatheterization was used to study the effect of renal arterial infusion of acetylcholine or secretin on medullary collecting duct function in anaesthetized rats. Acetylcholine infusion was associated with natriuresis and increased sodium delivery to, and decreased reabsorption in, the collecting duct. No changes from control function were found with secretin. Renal blood flow was increased with acetylcholine (+82%, p

Thiazide diuretic effect on medullary collecting duct function in the rat. The distal convoluted tubule is thought to be the principal site of action of thiazide diuretics, but, to our knowledge, there are no studies of their possible effects on collecting duct transport. Microcatheterization of the inner medullary collecting duct (IMCD) was carried out in rats undergoing a modest diuresis, natriuresis, and chloriuresis from hydrochlorothiazide (2 mg/kg/hr) and in normal controls. Delivery of fluid, sodium, and chloride to the beginning of the IMCD was increased, but not significantly, while the load remaining at the papillary tip (end) of the duct was increased markedly by hydrochlorothiazide. Chloride reabsorption in the IMCD was affected most markedly; the chloride reabsorption between the beginning and end of the duct, as a fraction of the delivered load, was reduced from 70.4 ± 5.4% in controls to insignificant amounts with hydrochlorothiazide (8.2 ± 11.5%, P < 0.001). The fraction of delivered sodium reabsorbed along the collecting duct was decreased from 78.5 ± 4.9% in controls to 37.2 ± 12.4% (P < 0.005) in thiazide-treated rats and fluid reabsorption was decreased from 59.4 ± 4.0% in controls to 31.9 ± 5.1% (P < 0.005). Small but significant potassium secretion into the IMCD occurred with hydrochlorothiazide, probably secondary to the marked increase in potassium delivery to the duct. Increased potassium excretion could account for a maximum of 50% of chloriuresis with hydrochlorothiazide. The observation that thiazide diuretics decrease chloride, sodium, and fluid reabsorption in the medullary collecting duct, like the recently demonstrated inhibitory effect of furosemide on this nephron segment, has significant implications for the rationale for diuretic use.Effet des diurétiques thiazidiques sur la fonction du canal collecteur médullaire chez le rat. Le tubule contourné distal est considéré comme étant le site principal d'action des diurétiques thiazidiques, mais, à notre connaissance, il n'existe aucune étude de leurs effets éventuels sur les transports dans le canal collecteur. Des microcathétérisations du canal collecteur médullaire interne (IMCD) ont été effectuées chez des rats ayant une diurèse, une natriurèse, et une chlorurèse modestes par l'hydrochlorothiazide (2 mg/kg/hr) et chez des rats contrôles. Le débit de liquide, de sodium, et de chlore au début de l'IMCD était augmenté, mais non significativement, tandis que la charge délivrée à l'extrémité (fin) papillaire du canal était augmentée de façon marquée par l'hydrochlorothiazide. La réabsorption de chlore dans l'IMCD était affectée de façon très marquée; la réabsorption de chlore entre le début et la fin du canal, sous forme de fraction de la charge délivrée, était réduite de 70,4 ± 5,4% chez les rats contrôles à des quantités insignifiantes par l'hydrochlorothiazide (8,2 ± 11,5%, P < 0,001). La fraction de sodium délivrée réabsorbée le long du canal collecteur était diminuée de 78,5 ± 4,9% chez les contrôles à 37,2 ± 12,4% (P < 0,005) chez les rats traités par le thiazide, et la réabsorption de liquide était diminuée de 59,4 ± 4,0% chez les contrôles à 31,9 ± 5,1% (P < 0,005). Une sécrétion faible mais significative de potassium dans l'IMCD est apparue avec l'hydrochlorothiazide, probablement secondaire à une augmentation marquée du potassium délivré au canal. L'augmentation de l'excrétion de potassium pourrait représenter au maximum 50% de la chlorurèse avec l'hydrochlorothiazide. L'observation que les diurétiques thiazidiques diminuent la réabsorption de chlore, de sodium et d'eau dans le canal collecteur médullaire, comme l'effet inhibiteur du furosémide récemment démontré dans ce segment néphronique, a des applications significatives pour l'utilisation rationnelle du diurétique.

Characteristics of sodium transport in the inner medullary collecting duct were determined in anesthetized rats before and during intravenous infusion of synthetic atrial natriuretic factor (atriopeptin II). Infusion of the factor was associated with increased sodium delivery and reduced fractional reabsorption in the duct. Increasing delivery to the same extent by KCl infusion had no effect on fractional reabsorption. The results demonstrate that atrial natriuretic factor has a specific inhibitory effect on net sodium transport in this part of the nephron. The mechanism of this inhibition may involve induction of sodium permeability and consequent backflux into the tubular lumen.

The effect of furosemide on inner medullary collecting duct chloride reabsorption has not been determined, and the blunting of furosemide action by drugs that inhibit prostaglandin synthesis, while known to occur, has not been examined in detail. The effect of indomethacin and meclofenamate on furosemide diuresis was studied in the rat using clearance and collecting duct microcatheterization methods. Furosemide-treated control animals showed complete inhibition of chloride, sodium, and water reabsorption in the inner medullary collecting duct. Rats given indomethacin or meclofenamate before and during furosemide administration showed marked reduction of the chloriuresis, natriuresis, and diuresis. Reduced delivery of sodium and chloride to the beginning of the inner medullary collecting duct, associated with a decrease in glomerular filtration rate and increased reabsorption in more proximal nephron segments, was largely responsible for the reduced natriuresis and chloriuresis during inhibition of prostaglandin synthesis. In addition, indomethacin increased collecting duct NaCl reabsorption toward normal, but meclofenamate showed no such effect. The results indicate that furosemide inhibits medullary collecting duct reabsorption of chloride, sodium, and water in the rat. The blunting of diuretic action seen with inhibition of prostaglandin synthesis is largely, although not entirely, due to effects of indomethacin and meclofenamate on furosemide action at nephron sites proximal to the collecting duct.

Collecting duct transport of fluid, sodium, and potassium was studied in rats infused with Ringer solution (5 ml·100 g body wt-1·h-1). A terminal segment of surface collecting duct in the exposed papilla was catheterized as far upstream as possible under visual observation. After fluid sampling the same duct was punctured at the same site with a glass micropipette and a second sample was taken. Samples were then obtained from the opening of the duct at the papilla tip by both catheter and micropipette. No significant difference between the two collection sites was found in the fraction of filtered sodium, potassium, or fluid remaining in the tubule, independent of the sampling technique used, indicating that volume expansion inhibited salt and water reabsorption. Although fractional fluid and sodium remainders were slightly higher and potassium remainder lower in upstream micropuncture samples compared to catheterization samples, the good correlation between collections obtained with both techniques suggests that both are equally valid as indicators of transport in terminal collecting ducts. Ringers infusion; extracellular fluid volume expansion; fluid reabsorption; sodium and potassium transport Submitted on May 7, 1979 Accepted on January 22, 1980

Tissue extracts derived from atria or ventricles of Sprague–Dawley rats were injected into Inactin-anesthetized assay rats. Compared with ventricular extracts, atrial extracts produced a 20 mmHg (1 mmHg = 133.322 Pa) fall in mean arterial blood pressure. This fall resulted from failure to increase cardiac output in compensation for peripheral vasodilation. Two factors were responsible: depression of heart rate (by 25 beats/min) and failure to increase cardiac performance. The time patterns and magnitudes of changes in cardiovascular parameters after cardiac extracts were not changed by prior atropinization. However, assay rats that were vagotomized showed no cardiac slowing after atrial extract and showed a significantly smaller decrease in mean arterial blood pressure than did sham-vagotomized or intact rats. Another group of assay rats was vagotomized as well as carotid-sinus-denervated before extract injection. In these rats the degree of hypotension caused by atrial extract was significantly greater than that observed after vagotomy alone and was not significantly different from that observed in rats with intact innervation. The results suggest that the hypotension that is caused by atrial extract, but not by ventricular extracts, results in part from the reflex effects of direct stimulation of chemosensitive cardiopulmonary receptors with vagal afferents and partly from the reflex effects of baroreceptor unloading. Ventricular extract had no hypotensive effect in any group of assay rats.

The aims of the present study were to examine urea handling along the length of the medullary collecting duct (MCD) in protein-depleted young rats and to determine the effect of urea infusion on MCD function and urine concentrating ability. In 10 young rats on a low protein diet, urea reabsorption equivalent to 18.3% of the filtered load was observed along the MCD (4.5 mm) using the microcatheterization technique. Collecting duct urea reabsorption occurred almost entirely (16.6%) in the distal portion of the MCD (mid-zone to papillary tip, 2.8 mm). These results are in contrast to the lack of net urea reabsorption along the MCD in protein-replete adult rats [21]. After urea infusion which raised plasma urea level from 3.5 to 10.5 mmol/l in protein-depleted rats, urine non-urea solute concentration increased in the non-exposed right kidney from 827 to 1,199 mosm kg−1 (P

Unabhangig vom Glomerulumfiltrat werden mit Hilfe einer Mikroperfusionsapparatur einzelne Abschnitte proximaler Konvolute der Rattenniere in situ perfundiert, um direkt am Orte der Sekretion den Transport von p-Aminohippursaure (PAH) zu untersuchen. Dabei wird einmal die Aufnahme von PAH aus dem Blut in die PAH-freie Perfusionslosung gemessen, zum anderen bei Tubulusperfusionen mit Losungen verschiedener PAH-Konzentrationen der Ausstrom in PAH-freies Blut.

SummaryNet fluxes of sodium, chloride and water across the ependymal lining of tral canal and that a sodium transport system is involved in this process.We are indebted to Dr. Leonard Napolitano of the Department of Anatomy for his help with the micrographs.

Medullary collecting duct function in the remnant kidney before and after volume expansion. The role of the medullary collecting ducts (CD) in the regulation of water and electrolyte excretion by the remnant kidney has not been determined. Medullary CD function was therefore studied by the microcathetenzation technique during hydropenia and after volume expansion with an isotonic saline load in rats with sham-operated normal kidney (stage I), remnant kidney (stage II), remnant kidney after contralateral nephrectomy (stage III), and sham-operated normal kidney after contralateral nephrectomy (stage-III control). Sodium, potassium, water, and solute reabsorption along the medullary CD were not altered in stage II during hydropenia when compared to normal control (stage I), but water reabsorption proximal to the medullary CD was decreased. In stage III, where the uremia was mild (BUN, 41 mg/dl), the fractions of the filtered load of sodium (72%) and water (62%) which were reabsorbed along the medullary CD were reduced in comparison to stage II (94% and 83%, respectively), stage I, or stage-III control. The fraction of filtered potassium entering the medullary CD was increased to 53% in stage III, compared to 16% in stage II, and 10 or 11% in stage I or stage-III control. No change in the fraction of filtered potassium remaining along the collecting ducts was observed. After volume expansion, there was no significant change in the fraction of filtered sodium and water remaining along the medullary CD in stage I, II, or III. The greater fractional excretion in stage III resulted from decreased reabsorption in more proximal nephron segments. The results indicate that (a) during hydropenia, fractional reabsorption of sodium and water is decreased along the medullary CD of the stage-III remnant kidney in the presence of mild uremia, (b) the increased fractional excretion of potassium in the remnant kidney with uremia is not determined by altered potassium handling in the medullary CD but occurs proximal to this nephron segment, and (c) extracellular fluid volume expansion with isotonic saline results in similar inhibition of fractional sodium and water reabsorption in the collecting ducts of both remnant and normal kidneys. Fonction du canal collecteur medullaire du rein reduit avant et apres expansion extra-cellulaire. Le role des canaux collecteurs (CD) medullaires dans la regulation de l'excretion de l'eau et des electrolytes par le rein reduit n'a pas encore ete precise. La fonction des CD medullaires a donc ete etudiee par microcatheterisation au cours de l'antidiurese et apres expansion au moyen de solute sale isotonique chez des rats dans les situations suivantes: rein intact ayant subi un simulacre de reduction (stade I), rein reduit (stade II), rein reduit et nephrectomie controlaterale (stade III), rein ayant subi un simulacre de reduction et nephrectomie controlaterale (temoin pour le stade III). La reabsorption d'eau, de sodium, de potassium, et d'autres substances dissoutes n'est pas modifiee, au cours de l'antidiurese, le long du CD medullaire des reins reduits de stade II par rapport aux controles normaux du stade I. Cependant la reabsorption d'eau en amont du CD medullaire est diminuee. Au stade III, ou l'uremie est moyenne (azote ureique, 41 mg/dl), les fractions des charges filtrees de sodium (72%) et d'eau (62%) qui sont reabsorbees le long du CD medullaire sont diminuees par rapport au stade II (94% et 83%, respectivement), ou par rapport aux stades I ou III-controle. La fraction du potassium filtre qui entre dans le CD medullaire est augmentee a 53% dans le stade III alors qu'elle represente 16% au stade II, 10 et 11% aux stades I et III-controle. Il n'a pas ete observe de modification de la fraction de potassium filtre encore presente dans les canaux collecteurs. Apres l'expansion il n'y a pas de modification significative de la fraction du sodium et de l'eau filtres presente le long des CD medullaires dans les stades I, II, et III. L'augmentation de l'excretion fractionnelle au stade III resulte d'une diminution de la reabsorption dans les segments plus proximaux du nephron. Les resultats indiquent que: (a) au cours de l'antidiurese la reabsorption fractionnelle du sodium et de l'eau est diminuee le long du CD medullaire du rein reduit au stade III en presence d'une uremie moyenne, (b) l'augmentation de l'excretion fractionnelle de potassium par le rein reduit au cours de l'uremie n'est pas liee a une modification du comportement du CD medullaire vis a vis du potassium mais survient en amont de ce segment, et (c) l'expansion determine une inhibition des reabsorptions fractionnelles de sodium et d'eau comparables dans les CD des reins reduits et des reins normaux.

Following the discovery of the natriuretic effect of atrial extract, our laboratory attempted to dissect the possible physiological role of atrial natriuretic factor. Initial micropuncture experiments demonstrated that the reduction of tubular sodium reabsorption was localized in the medullary collecting duct, a nephron site in which sodium transport was known to be inhibited after acute hypervolemia. Partial removal of the endogenous source of atrial natriuretic factor was associated with a reduced renal response to hypervolemia, confirming that the factor is causally involved in acute sodium balance. In vitro incubation of atrial tissue was used to investigate mechanisms of release of atrial natriuretic factor. It was found that agonists known to activate the intracellular polyphosphoinositide system in other tissues were effective in releasing natriuretic activity from the atria into the incubation medium. To determine whether atrial natriuretic factor might play a role in hypertension, atrial natriuretic content was measured in spontaneously hypertensive rats and their normotensive controls. Hypertension was associated with increased content. Since the renal response to exogenous factor was not impaired in these animals, we suggested that the increased content might play a compensatory role. Our early studies thus indicated that atrial natriuretic factor was a previously unrecognized hormone involved in cardiovascular regulation.

1. The possible involvement of renal nerves in the diuresis and natriuresis of blood volume expansion was studied in anaesthetized rats. Acute unilateral renal denervation caused increased excretion of fluid, sodium and potassium. 2. Renal blood and plasma flows were elevated without change in filtration rate. Intracortical blood flow distribution was not affected by the denervation. 3. Blood infusion caused diuresis, natriuresis and kaliuresis in both denervated and shamdenervated kidneys, associated with comparable initial increases in filtration and decreases in renal blood flow. No change in flow distribution was found, whether or not renal nerves were intact. 4. Although the magnitude of the excretory response to hypervolaemia was greater in denervated kidneys, the temporal pattern was identical with that of sham-operated kidneys. Our data thus do not show an effect of efferent renal nerve activity on volume natriuresis.

The microcatheterization technique was used to study transport of fluid, sodium, and potassium in the medullary collecting duct in rats (232–357 g) before and during administration of diuretic drugs. Series I received furosemide (1.5 mg initial dose + 1.5 mg/h), Series II acetazolamide (0.75 mg +0.75 mg/h), and Series III and IV mannitol at 0.3 g+0.3 g/h, and 0.3 g+1.0 g/h, respectively. All diuretics caused diuresis, natriuresis, and kaliuresis. The renal response to furosemide and acetazolamide was associated with increased hematocrit and decreased filtration rate, indicating depletion of blood volume. No such effect was seen in Series III and IV. In the collecting duct furosemide completely abolished the normal reabsorption of sodium and fluid and initiated net secretion of potassium. Partial inhibition of salt and water transport in the collecting duct was observed with acetazolamide and the low dose of mannitol (III). Both treatments resulted in potassium secretion. In Series IV the high rate of mannitol infusion was associated with complete inhibition of salt and water reabsorption from the medullary collecting system similar to that of Series I. The greater excretory response in this group compared to the furosemide series was due to increased delivery of tubular load to the collecting duct. It is concluded that a major site of action of furosemide is in the medullary duct, resulting in quantitative inhibition of salt and water reabsorption from this nephron segment. The partial transport inhibition during acetazolamide or modest mannitol diuresis can be explained by the presence of poorly absorbable solute in duct fluid. The mechanism of inhibition of reabsorption after the high rate of mannitol infusion remains undetermined.

Secretion of atrial natriuretic factor (ANF) in vivo is thought to be mediated by atrial distension. We have shown previously that nonstretched atria can release natriuretic activity in vitro when stimulated by certain agonists. In the present study atrial appendages from freshly excised rat hearts were incubated at 37 degrees C for up to 1 h in the presence of either vasopressin (5 X 10(-9) mol/l) or angiotensin II (2.5 X 10(-7) mol/l). Aliquots of postincubation media were injected intravenously into anesthetized bioassay rats to determine natriuretic activity. Control media, in which atria had been incubated without agonist, did not cause natriuresis. Significant increases in sodium excretion were seen after injection of media in which atria had been incubated in the presence of either agonist. Injection of medium with the same agonist concentration did not result in comparable natriuresis. Radioimmunoassay (RIA) indicated a high concentration of immunoactive ANF in the natriuretic media. However, radioreceptor assay (RRA) of the same media gave apparent ANF concentrations that were lower by about three orders of magnitude. Because the antibody used in the RIA cross reacts with ANF prohormone, whereas the RRA is sensitive only to the active form, we concluded that agonist-induced, stretch-independent release of ANF is in the form of prohormone, which can be converted to the active hormone in the circulation of the bioassay animal. The conclusion of prohormone release was confirmed by liquid chromatography. The data thus suggest that receptor-mediated as well as stretch-induced ANF secretion may be important in regulating the activity of the ANF system.

1. The purpose of this study was to evaluate the renal mechanisms which lead to a high urine [K+] in adrenalectomized (ADX) rats devoid of aldosterone. 2. By dividing the urine [K+] by the urine to plasma osmolality ratio, the [K+] in the cortical collecting duct luminal fluid can be estimated; dividing this value by the plasma [K+] yields an index of the transtubular [K+] gradient (TTKG) in vivo. 3. The TTKG was close to 7 in aldosterone deficient ADX rats while on a normal K+ diet and fell towards unity when amiloride or a low K+ diet was administered to these rats. 4. With a longer time on a low K+ diet, the TTKG was less than 1 in ADX rats. This suggests that K+ was reabsorbed in the medullary collecting duct under these conditions. 5. Hyperkalemia appears to have an ‘aldosterone-like’ action in the cortical collecting duct in vivo in the absence of aldosterone in ADX rats. This action of hyperkalemia permits normal K+ excretion rates despite the absence of mineralocorticoids.

Prostaglandin synthesis inhibition during volume expansion: Collecting duct function. The aim of this study was to examine the possible role of renal prostaglandins in the response of the inner medullary collecting duct (IMCD) to acute volume expansion. Collecting duct microcatheterization and clearance studies were carried out in anesthetized rats, volume-expanded with isotonic Ringer's solution. In volume-expanded control animals, there was no significant sodium or chloride reabsorption between the beginning and end (papillary tip) of the IMCD. Administration of indomethacin or meclofenamate prior to and during volume expansion in two other groups of rats resulted in significant water, sodium, and chloride reabsorption along the IMCD and markedly blunted the diuretic, natriuretic and chloriuretic response to volume expansion. Because delivery to the beginning of the duct was not significantly decreased, enhanced reabsorption in the IMCD largely accounted for the decrease in natriuresis and chloriuresis. Inner medullary tissue fluid chloride concentration increased after inhibition of prostaglandin synthesis. The results indicate that renal prostaglandins, perhaps by directly decreasing sodium chloride reabsorption, have an important role in the decrease in collecting duct reabsorption of sodium and chloride observed with acute volume expansion. L'inhibition de la synthese des prostaglandines pendant une expansion volemique: La fonction du tubule collecteur. Le but de cette etude a ete d'evaluer le role possible des prostaglandines renales dans la reponse du tube collecteur medullaire interne (IMCD) a une expansion volemique aigue. Une microcatheterisation des tubes collecteurs et des etudes de clairance ont ete faites chez des rats anesthesies, en expansion volemique par de la solution de Ringer isotonique. Chez les animaux controles en expansion volemique, il n'y avait pas de reabsorption significative du sodium ou du chlore entre le debut et la fin (pointe de la papille) de l'IMCD. L'administration d'indomethacine ou de meclofenamate avant et pendant une expansion volemique chez deux autres groupes de rats a entraine une reabsorption significative d'eau, de sodium et de chlore le long de l'IMCD et a inhibe de facon marquee la reponse diuretique, natriuretique et chloruretique a l'expansion volemique. Parce qu'il n'y avait pas de diminution significative des substances delivrees a l'entree du tube, l'augmentation de la reabsorption dans l'IMCD expliquait en grande partie la diminution de la natriurese et de la chlorurese. La concentration en chlore du liquide tissulaire de la medullaire interne s'est accrue apres inhibition de la synthese des prostaglandines. Ces resultats indiquent que les prostaglandines renales ont un role important dans la diminution de la reabsorption du sodium et du chlore dans le tube collecteur observe apres une expansion volemique aigue, peut-etre en diminuant directement la reabsorption de sodium chlore.

The microcatheterization technique was used to examine electrolyte transport in the medullary collecting duct of two groups of anesthetized rats during water diuresis and during a second experimental phase with 1-desamino-8-D-arginine vasopressin (dDAVP) administration or continued water diuresis. Potassium reabsorption of 53-61% of the delivered load was consistently observed in the medullary collecting duct during water diuresis. During dDAVP administration, urinary potassium excretion doubled, and there was no net potassium transport (reabsorption or secretion) in the medullary collecting duct. The change in potassium transport in medullary collecting duct from water diuresis to antidiuresis (dDAVP) was sufficient to account for the increase in urinary potassium excretion. Changes in flow rate, luminal sodium concentration, or collecting duct sodium reabsorption could not account for the changes in potassium transport in the collecting duct during dDAVP. The results are interpreted as indicating that dDAVP stimulates potassium entry (secretion) into the medullary collecting duct, probably by a direct effect. This action of antidiuretic hormone appears to be important in maintaining potassium homeostasis during changing water balance.

The distribution of 15-μm microspheres was measured in rat kidneys before and after the animals had undergone either isovolemic hematocrit increase or isovolemic hematocrit decrease. Raising the systemic arterial hematocrit from 46 ± 1% (mean ± SEM) to 59 ± 1% caused a significant decrease in the rates of urinary sodium and potassium excretion [Formula: see text], but no significant changes in total renal blood flow (RBF), filtration fraction (FF), outer cortical flow, inner cortical flow, or rate of urinary volume excretion [Formula: see text]. Fractional sodium excretion was also unchanged suggesting that the decreased [Formula: see text] was the result of a decreased glomerular filtration rate (GFR). The measured decrease in GFR was not statistically significant. Lowering the systemic hematocrit from 45 ± 1% to 33 ± 1% caused a significant fall in FF as well as significant increases in the rate of urine volume excretion [Formula: see text], [Formula: see text], and GFR. There was a significant increase in RBF but again no change in the flow distribution between inner and outer cortex. The findings show that hematocrit alterations alone do not immediately lead to a redistribution of blood flow between inner and outer cortex of the rat kidney.

The microsphere technique was used to measure renal blood flow and intrarenal flow distribution in cross-circulated pairs of rats. One rat of each pair was made hypervolemic by infusion of blood (2.3% of body weight), followed by intravenous reinfusion to urine to maintain intravascular expansion. The other rat of each pair, which also was urine-reinfused, was kept isvolemic throughout the experiment. As shown previously, blood infusion resulted in a large diuretic and natriuretic response in the hypervolemic partner, while a smaller but statistically significant response occurred in the isovolemic partner. Total renal blood flow did not change in the expanded rats and fell slightly but not significantly in the nonexpanded ones. Blood volume expansion was associated with a significant shift in microsphere concentration from outer to inner cortex. This shift was not, however, correlated with the magnitude of the renal response. The transferred natriuresis in isovolemic partners was not associated with any change in microsphere distribution. We conclude therefore, that redistribution of blood flow to the inner cortex, although a feature of intravascular expansion, is not the primary determinant of volume natriuresis. In addition, the natriuretic activity which develops in the blood of hypervolemic rats can reduce sodium reabsorption in isovolemic rats without intrarenal hemodynamic changes.

SUMMARY 1. Anaesthetized homozygous Brattleboro (DI) rats were used to study the renal response to iso-oncotic blood volume expansion. 2. With the same degree of hypervolaemia DI rats had exaggerated diuresis, natriuresis, and chloriuresis, but not kaliuresis, compared with heterozygous control rats. This increased excretion resulted in negative water balance by the end of the experiment in DI rats, whereas the controls showed restoration of normal balance. The control rats retained significant amounts of sodium and chloride, the Brattleboro rats, however, did not. 3. The lithium clearance method was used to localize the defect in sodium reabsorption. As judged from this method, there was a significantly lower sodium reabsorption in the proximal tubules as well as in the distal parts of the nephron of DI rats. 4. Plasma immunoreactivity of atrial natriuretic factor (ANF) was not different between groups before volume expansion. ANF levels rose and fell similarly in both groups during and after the iso-oncotic infusion. 5. Our results demonstrate that DI rats respond to acute hypervolaemia with an exaggerated diuresis and saluresis. The mechanism of the increased salt excretion may involve inhibition of sodium transport in the proximal tubules as well as in the distal parts of the nephron. These transport defects are not dependent on differing plasma ANF levels.