Analysis of energy metabolism and mechanism of loop diuretics in the thick ascending limb of Henle's loop in dog kidneys.

AIM: The thick ascending limb of Henle's loop (TALH) absorbs up to 40% of filtered NaCl in volume-expanded dogs. To examine if a fraction of this absorption is passive, NaHCO3 absorption and associated NaCl absorption in proximal tubules were inhibited by acetazolamide, a carbonic anhydrase inhibitor. RESULTS: Ouabain, a specific inhibitor of Na,K-ATPase activity, reduced the remaining NaCl absorption and renal oxygen consumption in a ratio DeltaNa/DeltaO2 = 18, as expected for active transport. However, the responses to two loop diuretics were DeltaNa/DeltaO2 = 24 for ethacrynic acid and DeltaNa/DeltaO2 = 30 for bumetanide. Both loop diuretics induced potassium secretion. By superimposing ouabain potassium secretion was stopped and DeltaNa/DeltaO2 = 18 restored. Replacement of half of the circulating NaCl with Na2SO4 gave stop-flow pattern similar to those obtained after ethacrynic acid. CONCLUSIONS: Low entry of some sodium ions thorugh the apical membrane is permitted despite low chloride supply or blockade by loop diuretics of chloride entry by the Na-K-2Cl transporter. Continued Na-K-ATPase activity causes secretion of potassium ions through the apical ion channel, ethacrynic acid being more kaliuretic and less natriuretic than bumetanide. Greater paracellular recycling of sodium ions after bumetanide maintains ionic balance. In contrast, under normal conditions excess entry of chloride by the Na-K-2Cl-transporter leads to paracellular back-diffusion of chloride rather than paracellular absorption of sodium ions, consistent with DeltaNa/DeltaO2 = 18 after ouabain. Thus all NaCl transport along TALH is active in vivo, whereas absorption of other cations, such as lithium, probably is passive.

Kinetic model of osmosis through semipermeable and solute-permeable membranes.

The gas analogy of the van't Hoff equation for osmotic pressure deltapi = RT/V, where R is gas constant, T absolute temperature and V mole volume of water, remained unexplained for a century because of a few misconceptions: (1) Use of supported membranes prevented the recognition that osmotic forces exert no effect on the solid membrane. During osmotic flow frictional force of solvent within membrane channels equals osmotic kinetic force pi at the interface against the solution containing impermeant solute. (2) Retrograde diffusion of water is much less than osmotic flow even when dx in the gradient dc/dx approaches zero. (3) The gas analogy was thought to be accidental. Actually, the internal kinetic pressure is P = RT/V, because intermolecular forces cancel out at the liquid interface, just as within a gas. The kinetic osmotic pressure is the difference in solvent pressure across the interface: pi = RT/V-(RT/V)X1 = (RT/V)X2, where X1 and X2 are the mole fractions of water and impermeant solute, respectively. Integration gives pi = -(RT/V)lnX1, identical to the thermodynamic equation. This equation is correct up to 25 atmospheres, and up to 180 atmospheres by assuming that a sucrose molecule binds 4 and a glycerol molecule 2.5 water molecules. For solute-permeable membranes, the reflection coefficient sigma can be calculated by formulas proposed for ultrafiltration. Because the fraction (1-sigma) of solute concentration behaves as solvent, osmosis may well proceed against the chemical potential gradient for water. The analogy to an ideal gas applies because pi = -(RT/V)lnX1 is the small difference between enormous internal solvent pressures.

Mechanisms of transjunctional transport of NaCl and water in proximal tubules of mammalian kidneys.

Tight junctions and the intercellular space of proximal tubules are not accessible to direct measurements of fluid composition and transport rates, but morphological and functional data permit analysis of diffusion and osmosis causing transjunctional NaCl and water transport. In the S2 segment NaCl diffuses through tight junctions along a chloride gradient, but against a sodium gradient. Calculation in terms of modified Nernst-Fick diffusion equation after eliminating electrical terms shows that transport rates (300-500 pmol min-1 mm-1 tubule length) and transepithelial voltage of +2 mV are in agreement with observations. Diffusion coefficients are Dtj=1500 microm2 s-1 in the S1 segment, and Dtj=90-100 microm2 s-1 in the S2 segment where apical intercellular NaCl concentration is 132 mM, 1 mM below complete stop (Dtj=0 and Donnan equilibrium). Tight junctions with gap distance 6 A are impermeable to mannitol (effective molecular radius 4 A); reflection coefficients are sigma=0.92 for NaHCO3 and sigma=0.28 for NaCl, because of difference in anion size. The osmotic force is provided by a difference in effective transjunctional osmolality of 10 mOsm kg-1 in the S1 segment and 30 mOsm kg-1 in the S2 segment, where differences in transjunctional concentration contribute with 21 mOsm kg-1 for NaHCO3 and -4 mOsm kg-1 for NaCl. Transjunctional difference of 30 mOsm kg-1 causes a volume flow of 2 nL min-1 mm-1 tubule length. Luminal mannitol concentration of 30 mM stops all volume flow and diffusive and convective transport of NaCl. In conclusion, transjunctional diffusion and osmosis along gradients generated by transcellular transport of other solutes account for all NaCl transport in proximal tubules.

Mechanisms of intercellular hypertonicity and isotonic fluid absorption in proximal tubules of mammalian kidneys.

The main purpose of this theoretical analysis (second of two articles) is to examine whether transjunctional diffusion of NaCl causes intercellular hypertonicity, which permits transcellular water transport across solute-impermeable lateral cell membranes until osmotic equilibration. In the S2 segment with tubular NaCl concentration 140 mM, the calculated apical intercellular NaCl concentration is c0 approximately 132 mM, which exceeds peritubular NaCl concentration by 12 mM or 22 mOsm kg-1. Variations in volume flow, junctional reflection coefficient (sigmaNaCl = 0.25-0.50), gap distance (g = 6-8 A), junctional depth (d = 18-100 A), intercellular diffusion coefficient (DLIS=500-1500 microm2 s-1) and hypothetical active NaCl transport alter c0 only by a fraction of 1 mM. However, dilution and back-leakage of NaHCO3 lower apical intercellular hyperosmolality to approximately 18 mOsm kg-1. Water transport through solute-impermeable lateral cell membranes continues until intercellular and cellular osmolalities are equal. Transcellular and transjunctional volume flow are of similar magnitude (2 nL min-1 mm-1 tubule length) in the S2 segment. Thus, diffusion ensures isotonic absorption of NaCl. Two-thirds of NaHCO3 and other actively transported sodium salts are extruded into the last third of the exponentially widening intercellular space where the exposure time is only 0.9 s. Osmotic equilibration is dependent on aquaporins in the cell membranes. If permeability to water is low, transcellular water transport stops; tubular fluid becomes hypotonic; NaCl diffusion diminishes, but transjunctional water transport remains unaltered as long as transcellular transport of NaHCO3 and other solutes provides the osmotic force.

Analysis of myogenic mechanisms in renal autoregulation.

To examine whether local myogenic mechanisms account for autoregulation of renal blood flow, a theoretical analysis was undertaken on a model of the pre-glomerular vascular tree consisting of a main and a short, narrow juxtaglomerular segment. At atmospheric extravascular pressure in vitro data are consistent with a relationship r=r0(1 + k - pk) between radius (r) and transmural pressure (p) at p > 60 mmHg, where k can be estimated from in vitro data and r=r0 at complete autoregulatory vasodilation. After introducing r=r(0)(1 + k - pk), Poiseuille's formula was integrated along the main segment, Deltax long, between arterial pressure P(1) and P(2) at the end of the main segment. At the lowest autoregulatory pressure P(1)=65 mmHg pre-glomerular blood flow is F=5Kr(0)(4)/Deltax. At P(1)=140 mmHg a pressure drop of only 17 mmHg to P2=123 mmHg is sufficient to fulfil the criterion for complete autoregulation: F=5Kr(0)(4)/Deltax. Thus, 80% of the total pre-glomerular vascular resistance is localized to the juxtaglomerular segment. Loop diuretics may abolish juxtaglomerular contractility. Calculated flow/pressure relationships after eliminating juxtaglomerular contractility are similar to those obtained after administering ethacrynic acid. If a constant tension hypothesis (r=60r(0)/p) rather than the transmural pressure hypothesis [r=r(0)(1 + k - pk)] applies, complete autoregulation is maintained to P(2)=89 mmHg, but the effect of loop diuretics is not mimicked. In conclusion, high juxtaglomerular contractility may be attributed to a myogenic mechanism only if extravascular pressure in the juxtaglomerular segment is subatmospheric.

Analysis of the tubuloglomerular feedback mechanism in renal autoregulation.

Along the juxtaglomerular segment of the afferent arteriole the luminal pressure p approaches the glomerular capillary pressure of 55-60 mmHg. At such low luminal pressures the myogenic mechanism contracts only if extravascular pressure p(ex) is subatmospheric. According to Poiseuille's formula complete autoregulation requires that blood flow is F=5Kr(0)(4)/Deltax at arterial pressures exceeding 65 mmHg; r(0) is the radius of the relaxed segment at transmural pressure p - p(ex) < or =60 mmHg, where p(ex) is the extravascular pressure; Deltax is the length of the main preglomerular segment, 10 times longer than the juxtaglomerular segment. Consistent with in vitro studies a myogenic mechanism may reduce the relaxed juxtaglomerular radius r(jx)=0.7r(0) by 40% at a transmural pressure of 140 mmHg. Fifty and 60% reductions are also considered. Integration of Poiseuille's formula shows that complete autoregulation of preglomerular blood flow requires negative extravascular pressures p(ex)= -90 to -55 mmHg dependent on contractile force. Negative pressure of this magnitude is generated by effective hyperosmolality <5 mOsm across the membrane separating cleft from pole cushion. Negative pressure stays constant at arterial pressures exceeding 90-110 mmHg, implying constant tubuloglomerular feedback, but approaches atmospheric pressure at lower arterial pressure, suggesting maintenance of blood flow by reduction in the glomerular filtration rate; a rise in macula densa concentrations [NaCl](md) by 0.15 mM or [NaHCO(3)](md) by 2 mM raises extravascular pressure towards atmospheric levels by approximately 40 mmHg. A 40-mmHg rise in interstitial pressure exerts the same effect. Loop diuretics nullify osmotic force and dilate juxtaglomerular and main segments by raising juxtaglomerular extravascular pressure towards atmospheric levels.

Inhibitory effect of endothelin on renin release in dog kidneys.

To investigate the effect of endothelin on renin release, experiments were performed in barbiturate-anaesthetized dogs with denervated kidneys. Intrarenal infusion of endothelin (1 ng min-1 kg-1 body wt) reduced renal blood flow (RBF) from 145 +/- 10 ml min-1 to 98 +/- 9 ml min-1 without altering renin release (1 +/- 1 microgram angiotensin I (AI) min-1). Renin release was then increased either by renal arterial constriction or ureteral occlusion. When renal arterial pressure was reduced to 50 mmHg, renin release averaged 79 +/- 20 micrograms AI min-1 in six dogs and fell significantly to 24 +/- 6 micrograms AI min-1 during endothelin infusion. During ureteral occlusion the inhibitory effect of endothelin on renin release either during inhibition of beta-adrenergic activity with propranolol or after inhibiting prostaglandin synthesis by indomethacin during intrarenal infusion of isoproterenol was examined. After propranolol administration ureteral occlusion increased renin release from 5 +/- 2 micrograms AI min-1 to 38 +/- 12 micrograms AI min-1 in six dogs. Subsequent intrarenal endothelin infusion (1 ng min-1 kg-1 body wt) during maintained ureteral occlusion reduced renin release to 10 +/- 3 micrograms AI min-1. In six other dogs prostaglandin synthesis was inhibited by indomethacin. Subsequent infusion of isoproterenol (0.2 microgram min-1 kg-1 body wt) to stimulate beta-adrenoceptor activity increased renin release from 13 +/- 4 micrograms AI min-1 to 68 +/- 8 micrograms AI min-1 during ureteral occlusion.(ABSTRACT TRUNCATED AT 250 WORDS)

Effect on renin release of inhibiting renal nitric oxide synthesis in anaesthetized dogs.

Nitric oxide plays an important role in the regulation of basal renal blood flow. This study was performed to examine whether selective inhibition of renal nitric oxide synthesis affects renin release in vivo. Accordingly, in six barbiturate-anaesthetized dogs renin release was examined before and after intrarenal infusion of the selective inhibitor of nitric oxide synthesis, NG-nitro-L-arginine (NOARG). NOARG was infused into the renal artery to yield a renal arterial blood concentration of 0.4 mumol ml-1. NOARG did not change systemic arterial blood pressure and glomerular filtration rate, but reduced basal renal blood flow by 26 +/- 2%. Urine flow, sodium and potassium excretion were reduced after inhibition of renal nitric oxide synthesis. Basal renin release (3 +/- 2 micrograms AI min-1) was not altered by NOARG infusion (1 +/- 1 micrograms AI min-1). To stimulate renin release the renal artery was constricted to a renal perfusion pressure of 50 mmHg. At this perfusion pressure infusion of NOARG reduced renin release significantly from 48 +/- 11 micrograms AI min-1 to 14 +/- 4 micrograms AI min-1. In conclusion, inhibition of renal nitric oxide synthesis reduces basal renal blood flow and reduces renin release stimulated by renal arterial constriction. These findings indicate that renal nitric oxide modulates both renal blood flow and renin release in vivo.

Plasma potassium concentration as a determinant of proximal tubular NaCl and NaHCO3 reabsorption in dog kidneys.

To examine whether an acute increase in plasma potassium concentration ([K]p) may inhibit proximal tubular transport, clearance studies were performed in seven anaesthetized, volume expanded dogs treated with amiloride (1 mg kg-1 body wt) to block tubular potassium secretion, and with bumetanide (30 micrograms kg-1 body wt) to inhibit NaCl reabsorption in Henle's loop. As [K]p was raised in steps from 2.6 +/- 0.2 to 7.9 +/- 0.2 mM, bicarbonate, chloride, and sodium reabsorption decreased by 232 +/- 56, 520 +/- 59 and 958 +/- 112 mumol min-1, respectively, at constant glomerular filtration rate (GFR). On average, the molar ratio between the inhibitory effects on bicarbonate and chloride reabsorption were 1:2.2-2.4. Reabsorption was calculated at GFR 100 ml min-1: (reabsorption 100/GFR (mmol min-1). It was inversely correlated to ln [K]p with r = -0.82 for bicarbonate and with r = -0.89 for chloride. Fractional potassium reabsorption remained constant at 0.31 +/- 0.03. Administration of acetazolamide (100 mg kg-1 body wt) in eight dogs at [K]p 8 mM reduced fractional reabsorption of bicarbonate, chloride and sodium as much as in previous studies on normokalaemic dogs. We conclude that acute elevation of [K]p inhibits NaHCO3 transport and passive proximal tubular NaCl reabsorption. This inhibition is not related to changes in potassium secretion and carbonic anhydrase activity, but may be secondary to depolarization of the basolateral membrane.

Luminal and basolateral uptake and degradation of insulin in the proximal tubules of the dog kidney.

In order to determine the major routes of insulin degradation in the body, insulin was labelled with a 'trapped' or 'residualizing' label: [125I]tyramine-cellobiose ([125I]TC) and injected intravenously in dogs. In contrast to conventional iodine-labelled insulin (131I-insulin), the [125I]TC-insulin allows measurements of total uptake in specific organs in vivo because the radioactive degradation products do not leave the cells. One h after the injection of trace doses, the amount of radioactivity recovered in the kidney from [125I]TC-insulin was nine times higher than when conventional [131I]insulin was used. In the blood, the amount of acid-precipitable radioactivity was the same for both labelled preparations, indicating similar clearance rates. A comparison of the uptake of insulin in filtering vs. non-filtering (ureter-occluded) kidneys indicated that the uptake of insulin is twice as high through the luminal than through the basolateral cell membrane; after 60 min, 8.9 +/- 0.8% of the injected [125I]TC-insulin dose remained in the filtering kidney and 3.2 +/- 0.2% of the dose was accumulated in the contralateral kidney, with occluded ureter but normal blood perfusion. In both filtering and non-filtering (ureter-occluded) kidneys, the subcellular distributions of [125I]TC-insulin were studied after various times by isopycnic sedimentation in sucrose gradients. No difference between peritubular and tubular uptake was discernible. The intracellular transport was rapid, leading to accumulation of radioactive label in dense lysosomes within 10 min.

Effect of atrial natriuretic factor on renal prostaglandin E2 release in the anaesthetized dog.

Experiments were undertaken in two groups of barbiturate anaesthetized dogs to examine whether atrial natriuretic factor (ANF) exerts an effect on renal release of prostaglandin E2 (PGE2). In the first group, intravenous infusion of ANF (50 ng min-1 kg-1 body wt) reduced basal PGE2 release from 4.4 +/- 0.8 pmol min-1 to 1.8 +/- 0.7 pmol min-1. In the second group, intrarenal infusion of an alpha 1-adrenoceptor agonist, phenylephrine (2.5-6.75 micrograms min-1), raised PGE2 release from 2.7 +/- 0.5 pmol min-1 to 7.5 +/- 1.3 pmol min-1. During continuous alpha 1-adrenergic stimulation, intravenous infusion of ANF (100 ng min-1 kg-1 body wt) reduced PGE2 release to 3.5 +/- 1.0 pmol min-1. These results demonstrate that ANF reduces basal and alpha 1-adrenergic stimulated renal PGE2 release.

Inhibition of renal nitric oxide synthesis with NG-monomethyl-L-arginine and NG-nitro-L-arginine.

In barbiturate-anesthetized dogs, the effects of intrarenal infusion of the two selective inhibitors of nitric oxide synthesis, NG-monomethyl-L-arginine (L-NMMA) and NG-nitro-L-arginine (NO-Arg), were compared. Basal renal blood flow (RBF) was reduced by 15 +/- 2% after L-NMMA at 0.28 mumol/ml, by 20 +/- 3% after NO-Arg at 0.07 mumol/ml, and by 31 +/- 5% after NO-Arg at 0.56 mumol/ml. Endothelium-dependent vasodilation induced by intrarenal infusion of acetylcholine was unaltered after L-NMMA, reduced by 24 +/- 3% after NO-Arg at 0.07 mumol/ml, and reduced by 59 +/- 13% after NO-Arg at 0.56 mumol/ml. Endothelium-independent vasodilation induced by intrarenal infusion of atrial natriuretic factor was not reduced after L-NMMA and NO-Arg. This study shows that NO-Arg is more potent than L-NMMA in inhibiting basal renal nitric oxide synthesis. In contrast to L-NMMA, NO-Arg exerted an inhibitory effect on acetylcholine-induced renal vasodilation. Our findings indicate that one-third of the basal RBF and more than one-half of the increase in RBF during acetylcholine infusion are dependent on nitric oxide synthesis.

Importance of nitric oxide in canine femoral circulation: comparison of two NO inhibitors.

OBJECTIVE: The aim was to assess the importance of endothelium derived nitric oxide (NO) in the regulation of vascular tone in the limbs. Changes in the canine femoral circulation were investigated after inhibition of NO synthesis. METHODS: The effects of two NO inhibitors, NG-monomethyl-L-arginine (LNMMA) and NG-nitro-L-arginine (NOARG), were compared on basal femoral blood flow and on endothelium dependent (acetylcholine) and endothelium independent (glyceryl trinitrate) vasodilatation in 15 pentobarbitone anaesthetised mongrel dogs. An electromagnetic flow probe was placed on the femoral artery to measure femoral blood flow. One catheter was advanced into the femoral artery proximal to the flow probe for blood pressure recording and another catheter distal to the flow probe for drug infusions. RESULTS: LNMMA (0.28 mumol.ml-1) reduced basal femoral blood flow by 44(SEM 3)%, NOARG (0.07 mumol.ml-1) by 21(4)%, and NOARG (0.56 mumol.ml-1) by 29(3)%. The flow responses to acetylcholine were reduced after LNMMA by 27(8)%, unaltered after NOARG (0.07 mumol.ml-1), and reduced after NOARG (0.56 mumol.ml-1) by 60(7)%. The flow response to glyceryl trinitrate was unaltered. L-arginine re-established femoral blood flow after infusion of LNMMA and NOARG (0.07 mumol.ml-1), but L-arginine did not re-establish femoral blood flow after NOARG (0.56 mumol.ml-1), even when infused in a 60-fold molar excess. CONCLUSIONS: There is a continuous basal release of NO in the canine femoral circulation. The results obtained by infusing LNMMA suggest that more than 40% of basal femoral blood flow is mediated by endothelium derived NO. Whereas LNMMA was more potent than NOARG in reducing basal NO release, NOARG (0.56 mumol.ml-1) reduced acetylcholine induced vasodilatation by as much as 60%.

Atrial natriuretic factor and renal sodium excretion during ventilation with PEEP in hypervolemic dogs.

Controlled mandatory ventilation with positive end-expiratory pressure (PEEP) reduces renal sodium excretion. To examine whether atrial natriuretic factor (ANF) is involved in the renal response to alterations in end-expiratory pressure in hypervolemic dogs, experiments were performed on anesthetized dogs with increased blood volume. Changing from PEEP to zero end-expiratory pressure (ZEEP) increased sodium excretion by 145 +/- 61 from 310 +/- 61 mumol/min and increased plasma immunoreactive (ir) ANF by 104 +/- 27 from 136 +/- 21 pg/ml. Changing from ZEEP to PEEP reduced sodium excretion by 136 +/- 36 mumol/min and reduced plasma irANF by 98 +/- 22 pg/ml. To examine a possible causal relationship, ANF (6 ng.min-1.kg body wt-1) was infused intravenously during PEEP to raise plasma irANF to the same level as during ZEEP. Sodium excretion increased by 80 +/- 36 from 290 +/- 78 mumol/min as plasma irANF increased by 96 +/- 28 from 148 +/- 28 pg/ml. We conclude that alterations in end-expiratory pressure lead to great changes in plasma irANF and sodium excretion in dogs with increased blood volume. Comparison of the effects of altering end-expiratory pressure and infusing ANF indicates that a substantial part of the changes in sodium excretion during variations in end-expiratory pressure can be attributed to changes in plasma irANF.

Effects of bradykinin and papaverine on renal autoregulation and renin release in the anaesthetized dog.

The present study on six anaesthetized dogs investigates the influences of two different vasodilators, bradykinin and papaverine, on the relationship between autoregulation of renal blood flow and glomerular filtration rate, sodium excretion and renin release. At control conditions renal blood flow and glomerular filtration rate was autoregulated to the same levels of renal arterial pressure, 55 +/- 3 and 58 +/- 3 mmHg, respectively. Renin release increased from 0.3 +/- 0.1 to 22 +/- 4 micrograms AI min-1, and sodium excretion decreased from 99 +/- 29 to 4.6 +/- 3.3 mumol min-1 when renal arterial pressure was reduced from 122 +/- 6 to 44 +/- 2 mmHg. Infusion of bradykinin (50 ng kg-1 min-1) increased renal blood flow by 50% at control blood pressure without changing glomerular filtration rate, and both renal blood flow and glomerular filtration rate autoregulated to the same pressure levels as during control. Sodium excretion increased threefold at control renal arterial pressure, but was unchanged at low renal arterial pressure. Bradykinin did not change renin release neither at control nor low renal arterial pressure. Papaverine infusion at a rate of 4 mg min-1 increased renal blood flow 50% without changing glomerular filtration rate. The lower pressure limits of renal blood flow and glomerular filtration rate autoregulation were increased to 94 +/- 6 and 93 +/- 6 mmHg, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)

Atrial natriuretic factor reduces renin release by opposing alpha-adrenoceptor activity.

To examine how atrial natriuretic factor (ANF) inhibits renin release during renal sympathetic nerve stimulation, experiments were performed in barbiturate-anesthetized dogs. In five dogs, intravenous ANF infusion (50 ng.min-1.kg body wt-1) reduced renin release induced by renal nerve stimulation (1 Hz) from 16.8 +/- 8.4 to 3.5 +/- 2.1 micrograms angiotensin I (ANG I)/min. In two groups, renin release was raised by ureteral occlusion, which enhances the effects of beta-adrenoceptor stimulation and increased prostaglandin synthesis. During ureteral occlusion, intrarenal infusion of isoproterenol (0.2 micrograms.min-1.kg body wt-1) increased renin release in eight dogs to 82.6 +/- 10.9 micrograms ANG I/min, which was not significantly reduced by ANF infusion (81.1 +/- 10.1 micrograms ANG I/min). Similarly, intrarenal infusion of arachidonic acid (80 micrograms.min-1.kg body wt-1) during ureteral occlusion increased renin release in five dogs to 22.2 +/- 3.0 micrograms ANG I/min, which was not significantly reduced by ANF infusion (22.5 +/- 3.5 micrograms ANG I/min). Finally, in six dogs examined at free urine flow, intrarenal infusion of phenylephrine, an alpha-adrenergic agonist, raised renin release from 0.5 +/- 0.3 to 20.1 +/- 6.8 micrograms ANG I/min, which was reduced to 10.6 +/- 3.9 micrograms ANG I/min by intravenous ANF infusion (100 ng.min-1.kg body wt-1). These results indicate that ANF does not counteract stimulation of renin release by beta-adrenoceptors and prostaglandins but reduces nerve-stimulated renin release by opposing alpha-adrenoceptor activity.

Release of atrial natriuretic factor during selective cardiac alpha- and beta-adrenergic stimulation, intracoronary Ca2+ infusion, and aortic constriction in pigs.

The effects of alpha- and beta-adrenergic stimulation on release of atrial natriuretic factor (ANF) were examined in seven anesthetized, open-chest pigs. The alpha-adrenergic agonist phenylephrine (28.0 micrograms/min) and the beta-adrenergic agonist isoproterenol (0.3 micrograms/min) were infused into the proximal part of the circumflex coronary artery to stimulate the left atrial adrenoceptors without concomitant changes in left and right atrial filling pressures (v wave). Isoproterenol reduced plasma immunoreactive ANF (irANF) by 15 +/- 7 pg/ml (20%) from 76 +/- 10 pg/ml despite a rise in left atrial systolic pressure (a wave). A comparable rise in left atrial systolic pressure, induced by intracoronary infusion of calcium chloride (8.0 mg/min), increased plasma irANF by 33 +/- 10 pg/ml (53%) from 62 +/- 7 pg/ml. Phenylephrine increased plasma irANF by 9 +/- 4 pg/ml (14%) from 66 +/- 10 pg/ml without altering right and left atrial pressures. A rise in left atrial filling pressure of 3.2 +/- 0.5 mm Hg, induced by constricting the ascending aorta, increased plasma irANF by 83 +/- 35 pg/ml (141%) from 59 +/- 11 pg/ml. This increase was nine times that during phenylephrine infusion. In conclusion, alpha-adrenergic stimulation increases and beta-adrenergic stimulation inhibits ANF release by a direct action on the atrial myocytes. The direct effects of alpha- and beta-adrenergic stimulation on ANF release in vivo are small compared with the effect of a moderate rise in atrial filling pressure.

Loop diuretics reduce lithium reabsorption without affecting bicarbonate and phosphate reabsorption.

The effects of the loop diuretics ethacrynic acid and bumetanide on lithium, bicarbonate and phosphate reabsorption were compared in 16 anaesthetized, normovolaemic dogs. In six dogs, ethacrynic acid (3 mg kg-1 body wt) significantly reduced absolute lithium reabsorption from 29.3 +/- 4.1 to 19.0 +/- 3.4 mumol min-1, fractional lithium reabsorption from 0.65 +/- 0.04 to 0.37 +/- 0.04 and fractional chloride reabsorption from 1.00 +/- 0.00 to 0.65 +/- 0.02. Bicarbonate and phosphate reabsorption did not decrease significantly. In six other dogs, bumetanide (30 micrograms kg-1 body wt) gave similar results. Absolute lithium reabsorption significantly decreased from 34.0 +/- 2.2 to 18.1 +/- 2.6 mumol min-1 and fractional lithium reabsorption decreased from 0.50 +/- 0.03 to 0.25 +/- 0.03. Fractional chloride reabsorption decreased from 0.98 +/- 0.00 to 0.61 +/- 0.05, whereas bicarbonate and phosphate reabsorption were not significantly altered. Thus, both loop diuretics greatly reduced lithium reabsorption. We propose that loop diuretics inhibit passive lithium reabsorption in the thick ascending limb of Henle's loop by reducing the lumen-positive electrical potential that drives passive cation transport.

Stimulation of renin release by PGE2 and PGI2 infusion in the dog: enhancing effect of ureteral occlusion or administration of ethacrynic acid.

This study on 19 anaesthetized dogs had two objectives. The first was to compare the potencies of PGE2 and PGI2 as stimulators of renin release and demonstrate their dependency on activation of intrarenal mechanisms for renin release. The second objective was to demonstrate that ethacrynic acid (ECA) increases renin release not as a stimulator, but by activating intrarenal mechanisms. After inhibiting renal prostaglandin synthesis by indomethacin, PGE2 and PGI2 infused into the aorta proximal to the renal arteries exerted no significant effects on renin release, but increased renin release during ureteral occlusion. At equimolar infusion rates, PGI2 increased renin release twice as much as PGE2, but this difference in potency may reflect differences in degradation since 86% of PGE2 and 29% of PGI2 (measured as 6-keto-PGF1 alpha) were degraded during one passage through the kidney. By infusing PGF2 at 8 nmol min-1 and PGI2 at 2 nmol min-1 renin release increased equally and the prostaglandin outputs increased to the same levels as during ureteral occlusion before indomethacin administration. ECA did not increase renin release after indomethacin administration. However, infusion of PGE2 during continuous ECA administration increased renin release in a dose-dependent manner similar to the experiments performed during ureteral occlusion. We conclude that PGI2 and PGE2 in the amounts synthesized in the kidney seem to be equally important stimulators of renin release but their relative potencies cannot be determined because the site of degradation is uncertain. Renin release is enhanced by intrarenal mechanisms activated by ECA infusion or ureteral occlusion, which both cause autoregulatory vasodilation and reduce NaCl reabsorption at the macula densa.