Tubular Outflow Research Articles

Developmental Renal Glomerular Defects at the Origin of Glomerulocystic Disease

The architecture of renal glomeruli is acquired through intricate and still poorly understood developmental steps. In our study we identify a crucial glomerular morphogenetic event in nephrogenesis that drives the remodeling/separation of the prospective vascular pole (the future entrance of the glomerular arterioles) and the urinary pole (the tubular outflow). We demonstrate that this remodeling is genetically programmed. In fact, in mouse and human, the absence of HNF1B impairs the remodeling/separation of the two poles, leading to trapping and constriction of the tubular outflow inside the glomerulus. This aberration gives rise to obstructive glomerular dilations upon the initiation of primary urine production. In this context, we show that pharmacological decrease of glomerular filtration significantly contains cystic expansion. From a developmental point of view, our study discloses a crucial event on glomerular patterning affecting the "inside-outside" fate of the epithelia in the renal glomerulus.

Gore-Tex Mesh as Cause for Distal Outflow Graft Obstruction in Ventricular Assist Device Patients

Purpose Re-do sternotomy following left ventricular assist device (LVAD) implant can occur due to cardiac transplantation or device complication. A polytetrafluoroethylene membrane (Gore-Tex) has been used to cover portions of the outflow graft to reduce adhesions. We describe 3 cases of late-presenting extrinsic compression of the distal outflow graft from a Gore-Tex membrane in both HVAD and HMII devices. Methods We conducted a retrospective analysis of all LVAD implants which included a Gore-Tex membrane at our institution between 1/2012-6/2018. We defined cases of extrinsic outflow graft obstruction (OGO) by clinical presentation, CT imaging, and intraoperative findings. Results During the study period, 158 LVADs with tubular Gore-Tex outflow graft coverage were implanted (93 HVAD, 59 HM2, 4 HM3). Three cases of extrinsic OGO (1 HVAD, 2 HM2) were identified, for a cumulative incidence of 1.9% (1.44 per 100 patient years). The average time to clinical presentation was 1157 ± 155 days. Each case presented with persistent low-flow alarms detected via remote monitoring in the absence of power spikes or clinical symptoms. Markers of hemolysis (LDH, plasma free hemoglobin) were normal, INR was therapeutic, and echocardiograms did not indicate volume depletion or right ventricular failure. In each case, CT imaging demonstrated critical narrowing of the distal outflow graft (N=2) or aortic anastomosis (N=1) (Fig A). Each patient underwent surgical exploration; upon incision of the overlying Gore-Tex membrane, a proteinaceous material was drained resulting in immediate improvement in flows and avoidance of pump exchange (Fig B). Conclusion We highlight 3 cases of a previously undescribed mechanism of OGO by extrinsic compression, resulting from accumulation of proteinaceous material underneath the Gore-Tex membrane. Each presented with gradual onset of low flows without additional symptoms, suggesting that a high level of clinical suspicion is necessary to ensure appropriate management.

Vangl2-Regulated Polarisation of Second Heart Field-Derived Cells Is Required for Outflow Tract Lengthening during Cardiac Development

Planar cell polarity (PCP) is the mechanism by which cells orient themselves in the plane of an epithelium or during directed cell migration, and is regulated by a highly conserved signalling pathway. Mutations in the PCP gene Vangl2, as well as in other key components of the pathway, cause a spectrum of cardiac outflow tract defects. However, it is unclear why cells within the mesodermal heart tissue require PCP signalling. Using a new conditionally floxed allele we show that Vangl2 is required solely within the second heart field (SHF) to direct normal outflow tract lengthening, a process that is required for septation and normal alignment of the aorta and pulmonary trunk with the ventricular chambers. Analysis of a range of markers of polarised epithelial tissues showed that in the normal heart, undifferentiated SHF cells move from the dorsal pericardial wall into the distal outflow tract where they acquire an epithelial phenotype, before moving proximally where they differentiate into cardiomyocytes. Thus there is a transition zone in the distal outflow tract where SHF cells become more polarised, turn off progenitor markers and start to differentiate to cardiomyocytes. Membrane-bound Vangl2 marks the proximal extent of this transition zone and in the absence of Vangl2, the SHF-derived cells are abnormally polarised and disorganised. The consequent thickening, rather than lengthening, of the outflow wall leads to a shortened outflow tract. Premature down regulation of the SHF-progenitor marker Isl1 in the mutants, and accompanied premature differentiation to cardiomyocytes, suggests that the organisation of the cells within the transition zone is important for maintaining the undifferentiated phenotype. Thus, Vangl2-regulated polarisation and subsequent acquisition of an epithelial phenotype is essential to lengthen the tubular outflow vessel, a process that is essential for on-going cardiac morphogenesis.

Erythropoietin down‐regulates proximal renal tubular reabsorption and causes a fall in glomerular filtration rate in humans

Recombinant human erythropoietin (rHuEPO) elevates haemoglobin concentration both by increasing red blood cell volume and by a decrease in plasma volume. This study delineates the association of rHuEPO-induced changes in blood volumes with changes in the renin–aldosterone system and renal function. Sixteen healthy males were given rHuEPO for 28 days in doses raising the haematocrit to 48.3±4.1%.Renal clearance studieswith urine collections (N = 8) were done at baseline and at days 4, 11, 29 and 42. Glomerular filtration rate (GFR) was measured by 51Cr-EDTA.Renal clearance of lithium (CLi)was used as an index of proximal tubular outflow and to assess segmental renal tubular handling of sodium and water. rHuEPO-induced increases in haematocrit occurred from day 10 onwards and was caused by both an increase in red cell volume and a fall in plasma volume. Well before that (from day 2 and throughout the treatment time), rHuEPO decreased plasma levels of renin and aldosterone (N = 8) by 21–33% (P < 0.05) and 15–36% (P < 0.05), respectively. After cessation of rHuEPO, values returned to baseline. On days 11 and 29, CLi increased (P < 0.02) indicating a significant 10–16% decrease in absolute proximal reabsorption of sodium and water (APR = GFR − CLi, P < 0.05). GFR decreased slightly, albeit significantly, on day 4 (P < 0.05). In conclusion, rHuEPO promptly, and before any changes in blood volumes and haematocrit can be detected, causes a down-regulation of the renin–aldosterone system. The results are compatible with a rHuEPO-induced reduction in proximal reabsorption rate leading to activation of the tubuloglomerular feedback mechanism and a fall in GFR. Therefore, treatment with rHuEPO may result in suppression of endogenous EPO synthesis secondary to a decrease in intrarenal oxygen consumption.

AVP-stimulated nucleotide secretion in perfused mouse medullary thick ascending limb and cortical collecting duct

Extracellular nucleotides are local, short-lived signaling molecules that inhibit renal tubular transport via both luminal and basolateral P2 receptors. Apparently, the renal epithelium itself is able to release nucleotides. The mechanism and circumstances under which nucleotide release is stimulated remain elusive. Here, we investigate the phenomenon of nucleotide secretion in intact, perfused mouse medullary thick ascending limb (mTAL) and cortical collecting duct (CCD). The nucleotide secretion was monitored by a biosensor adapted to register nucleotides in the tubular outflow. Intracellular Ca(2+) concentration ([Ca(2+)](i)) was measured simultaneously in the biosensor cells and the renal tubule with fluo 4. We were able to identify spontaneous tubular nucleotide secretion in resting perfused mTAL. In this preparation, 10 nM AVP and 1-desamino-8-D-arginine vasopressin (dDAVP) induced robust [Ca(2+)](i) oscillations, whereas AVP in the CCD induced large, slow, and transient [Ca(2+)](i) elevations. Importantly, we identify that AVP/dDAVP triggers tubular secretion of nucleotides in the mTAL. After addition of AVP/dDAVP, the biosensor registered bursts of nucleotides in the tubular perfusate, corresponding to a tubular nucleotide concentration of approximately 0.2-0.3 microM. A very similar response was observed after AVP stimulation of CCDs. Thus AVP stimulated tubular secretion of nucleotides in a burst-like pattern with peak tubular nucleotide concentrations in the low-micromolar range. We speculate that local nucleotide signaling is an intrinsic feedback element of hormonal control of renal tubular transport.

AVP-stimulated nucleotide secretion in perfused mouse medullary thick ascending limb and cortical collecting duct

Extracellular nucleotides are local, short-lived signaling molecules that inhibit renal tubular transport via both luminal and basolateral P2 receptors (1, 2). Apparently, the renal epithelium itself is able to release nucleotides (3, 4). The mechanism and circumstances under which epithelia nucleotide release is stimulated remains elusive (5, 6). Here, we investigate the phenomenon of nucleotide secretion in intact perfused mouse medullary thick ascending limb (mTAL) and cortical collecting duct (CCD). The nucleotide secretion was monitored by a biosensor cell placed to register nucleotides in the tubular out-flow. [Ca(2+)](i) was measured simultaneously in the biosensor cells and the renal tubule with fluo-4. We were able to identify spontaneous tubular nucleotide secretion in resting perfused mTAL. This was seen as lively [Ca(2+)](i) oscillations in the nucleotide biosensor cells when the tubular outflow fluid engulfed the sensing cells. In mouse mTAL 10 nM AVP and dDAVP induced robust [Ca(2+)](i) oscillations, whereas AVP in the CCD induced large, slow and transient [Ca(2+)](i) elevations. Importantly, we identify that AVP/dDAVP triggers tubular secretion of nucleotides in mTAL. After addition of AVP/dDAVP the biosensor cells registered bursts of nucleotides originating from the tubular perfusate. The approximated tubular nucleotide concentration reached peak values of approximately 0.2-0.3 microM. A very similar response was observed after AVP stimulation of CCDs. Thus, AVP stimulated tubular secretion of nucleotides in a burst like pattern with peak tubular nucleotide concentrations in the low micromolar range. Luminal nucleotides are prone to activate luminal P2 receptors which in turn are well described to inhibit AVP-augmented aquaporin-2-dependent water absorption or ENaC-mediated Na(+) transport (8). Therefore, we speculate that local nucleotide signaling is an intrinsic feed-back element of hormonal control of renal tubular transport.

Lipopolysaccharide-Induced Acute Renal Failure in Conscious Rats: Effects of Specific Phosphodiesterase Type 3 and 4 Inhibition

In conscious, chronically instrumented rats we examined 1) renal tubular functional changes involved in lipopolysaccharide (LPS)-induced acute renal failure; 2) the effects of LPS on the expression of selected renal tubular water and sodium transporters; and 3) effects of milrinone, a phosphodiesterase type 3 (PDE3) inhibitor, and Ro-20-1724, a PDE4 inhibitor, on LPS-induced changes in renal function. Intravenous infusion of LPS (4 mg/kg b.wt. over 1 h) caused an immediate decrease in glomerular filtration rate (GFR) and proximal tubular outflow without changes in mean arterial pressure (MAP). LPS-induced fall in GFR and proximal tubular outflow were sustained on day 2. Furthermore, LPS-treated rats showed a marked increase in fractional distal water excretion, despite significantly elevated levels of plasma vasopressin (AVP). Semiquantitative immunoblotting showed that LPS increased the expression of the Na(+),K(+),2Cl(-)-cotransporter (BSC1) in the thick ascending limb, whereas the expression of the AVP-regulated water channel aquaporin-2 in the collecting duct (CD) was unchanged. Pretreatment with milrinone or Ro-20-1724 enhanced LPS-induced increases in plasma tumor necrosis factor-alpha and lactate, inhibited the LPS-induced tachycardia, and exacerbated the acute LPS-induced fall in GFR. Furthermore, Ro-20-1724-treated rats were unable to maintain MAP. We conclude 1) PDE3 or PDE4 inhibition exacerbates LPS-induced renal failure in conscious rats; and 2) LPS treated rats develop an escape from AVP in the CDs, which could be aimed to protect against water intoxication in septic conditions associated with decreased GFR and high levels of AVP.

Glomerular and Tubular Function in Renal Transplant Patients Treated with and without Ciclosporin A

The present study evaluated whether chronically administered low-dose (<5 mg/kg) ciclosporin A (CsA) affects renal haemodynamics and tubular function in renal transplant recipients (RTx) when studied at nadir CsA blood levels. The renal clearance of lithium was used as an index of proximal tubular outflow of sodium and water. Effective renal plasma flow, glomerular filtration rate, and renal clearance of lithium were studied in 67 stable non-diabetic RTx and 44 healthy controls. Forty-eight of the RTx were treated with CsA, prednisone, and azathioprine. Nineteen were treated exclusively with prednisone and azathioprine. In RTx with a good graft function (serum-creatinine <125 µmol/l), no specific CsA-induced renal haemodynamic and tubular dysfunctions were evident. In CsA-treated RTx with a slightly reduced renal function (serum creatinine 125–180 µmol/l) a decrease in fractional proximal tubular reabsorption was found. The renal clearances of urate and magnesium were comparable between RTx treated with or without CsA, and a significant correlation between glomerular filtration rate and renal clearance of urate was found. CsA-treated RTx had a significantly higher blood pressure, independent of glomerular filtration rate and segmental tubular function. In conclusion, at nadir CsA blood levels, no specific CsA-induced tubular dysfunction evaluated by the renal lithium clearance method could be demonstrated in RTx receiving chronically low-dose CsA. The hyperuricaemia commonly seen in RTx seems to be mainly caused by the reduced glomerular filtration rate.

Developmental remodeling and shortening of the cardiac outflow tract involves myocyte programmed cell death.

The embryonic outflow tract is a simple tubular structure that connects the single primitive ventricle with the aortic sac and aortic arch arteries. This structure undergoes a complex sequence of morphogenetic processes to become the portion of the heart that aligns the right and left ventricles with the pulmonary artery and aorta. Abnormalities of the outflow tract are involved in many clinically significant congenital cardiac defects; however, the cellular and molecular processes governing the development of this important structure are incompletely understood. Histologic and tissue-tagging studies indicate that the outflow tract tissues compact and are incorporated predominantly into a region of the right ventricle. The hypothesis tested in the current study was that cell death or apoptosis in the muscular portion of the outflow tract is an important cellular mechanism for outflow tract shortening. The tubular outflow tract myocardium was specifically marked by infecting myocytes of the chicken embryo heart with a recombinant replication-defective adenovirus expressing beta-galactosidase (beta-gal) under the control of the cytomegalovirus promoter. Histochemical detection of the beta -gal-labeled outflow tract myocytes revealed that the tubular structure shortened to become a compact ring at the level of the pulmonic infundibulum over several days of development (stages 25-32, embryonic days 4-8). The appearance of apoptotic cardiomyocytes was correlated with OFT shortening by two histologic assays, TUNEL labeling of DNA fragments and AnnexinV binding. The rise and fall in the number of apoptotic myocytes detected by histologic analyses paralleled the change in activity levels of Caspase-3, a protease in the apoptotic cascade, measured in outflow tract homogenates. These results suggest that the elimination of myocytes by programmed cell death is one mechanism by which the outflow tract myocardium remodels to form the proper connection between the ventricular chambers and the appropriate arterial trunks.

Renal haemodynamics, sodium and water reabsorption during continuous intravenous infusion of recombinant interleukin-2

1. Renal haemodynamics, lithium and sodium clearance were measured in 14 patients treated with recombinant interleukin-2 for metastatic renal cell carcinoma. 2. Patients were studied before and after 72 h of continuous intravenous infusion of recombinant interleukin-2 (18x10(6) i.u..24 h-1.m-2) and 48 h post therapy. Cardiac output was measured by impedance cardiography. Effective renal plasma flow and glomerular filtration rate were determined by the renal clearances of 131I-hippuran and 99mTc-diethylenetriaminepenta-acetic acid (DTPA) respectively. Renal clearance of lithium (CLi) was used as an index of proximal tubular outflow. 3. Treatment caused a transient decrease in mean arterial blood pressure and systemic vascular resistance, but cardiac output remained unchanged. Renal blood flow decreased and renal vascular resistance increased during and after treatment. Sodium clearance decreased from 1.10 (0.63/1.19) ml/min to 0.17 (0.18/0.32) ml/min (P=0.003). Glomerular filtration rate remained unchanged, whereas the median CLi decreased from 26 (17/32) ml/min to 17 (10/21) ml/min (P=0.008). Calculated absolute proximal reabsorption rate of water increased from 63 (40/69) ml/min to 71 (47/82) ml/min (P=0.04). The urinary excretion rate of thromboxane B2 and the ratio between excretion rates of thromboxane B2 and 6-keto-prostaglandin-F1alpha increased by 98% (P=0.022) and 175% (P=0.022) respectively. 4. The study suggests a specific recombinant interleukin-2-induced renal vasoconstrictor effect. Changes in renal prostaglandin synthesis may contribute to the decrease in renal blood flow. The lithium clearance data suggest that an increased proximal tubular reabsorption rate may contribute to the decreased sodium clearance during recombinant interleukin-2 treatment.

Renal haemodynamics, sodium and water reabsorption during continuous intravenous infusion of recombinant interleukin-2

1.Renal haemodynamics, lithium and sodium clearance were measured in 14 patients treated with recombinant interleukin-2 for metastatic renal cell carcinoma.2.Patients were studied before and after 72 h of continuous intravenous infusion of recombinant interleukin-2 (18×106 i.u.·24 h-1·m-2) and 48 h post therapy. Cardiac output was measured by impedance cardiography. Effective renal plasma flow and glomerular filtration rate were determined by the renal clearances of 131I-hippuran and 99mTc-diethylenetriaminepenta-acetic acid (DTPA) respectively. Renal clearance of lithium (CLi) was used as an index of proximal tubular outflow.3.Treatment caused a transient decrease in mean arterial blood pressure and systemic vascular resistance, but cardiac output remained unchanged. Renal blood flow decreased and renal vascular resistance increased during and after treatment. Sodium clearance decreased from 1.10 (0.63/1.19) ml/min to 0.17 (0.18/0.32) ml/min (P = 0.003). Glomerular filtration rate remained unchanged, whereas the median CLi decreased from 26 (17/32) ml/min to 17 (10/21) ml/min (P = 0.008). Calculated absolute proximal reabsorption rate of water increased from 63 (40/69) ml/min to 71 (47/82) ml/min (P = 0.04). The urinary excretion rate of thromboxane B2 and the ratio between excretion rates of thromboxane B2 and 6-keto-prostaglandin-F1α increased by 98% (P = 0.022) and 175% (P = 0.022) respectively.4.The study suggests a specific recombinant interleukin-2-induced renal vasoconstrictor effect. Changes in renal prostaglandin synthesis may contribute to the decrease in renal blood flow. The lithium clearance data suggest that an increased proximal tubular reabsorption rate may contribute to the decreased sodium clearance during recombinant interleukin-2 treatment.

Proton secretion in the upper part of the descending limb of long-looped nephron from hamsters.

The proton transport processes in the upper part of the descending limb of the long-looped nephron (LDLu) from hamsters were studied using a fluorescent dye, 2',7'-bis(carboxyethyl)carboxyfluorescein (BCECF) in microperfused single nephron preparations. Intracellular pH (pHi), as assessed by the measurement of the fluorescence of BCECF trapped in the cytoplasm, was 7.23 +/- 0.05 (n = 18) under nominally HCO3--free conditions. Ouabain, when added to the bath, decreased pHi by 0.22 units. After an NH4Cl prepulse, the initial proton extrusion rate was 1.23 +/- 0.26 (n = 9) pH units/min, and was retarded in the presence of 1 mM amiloride either in the bath or in the lumen. pHi failed to recover when Na+ was eliminated from ambient solutions. These observations suggest that Na+/H+ antiporters exist both in the apical and basolateral cell membranes. By measuring tubular fluid pH (pHt) under stopped flow conditions, we examined whether the hamster LDLu has the capacity to generate and maintain a transmural H+ gradient. After the tubular outflow was obstructed, the luminal fluid was rapidly acidified, reaching a steady-state pH of 6.84 +/- 0.09 (n = 7). The steady-state pH was influenced by bath pH. Tubular fluid acidification was not observed in the absence of Na+ and was prevented by ouabain. We conclude that the hamster LDLu has the capability to generate and maintain a transmural proton gradient by proton secretion via a luminal Na+/H+ antiporter which is secondarily driven by the Na+-K+ ATPase in the basolateral membrane.

Dopamine natriuresis in salt‐repleted, water‐loaded humans: a dose‐response study

Aims The purpose of the present study was to define the dose‐response relationship between exogenous dopamine and systemic haemodynamics, renal haemodynamics, and renal excretory function at infusion rates in the range 0 to 12.5 μg kg−1 min−1 in normal volunteers. Methods While undergoing water diuresis, eight subjects were infused with 0, 1, 2, 3, 5, 7.5, 10 or 12.5 μg of dopamine kg−1 min−1 over 2 h in a randomized and double‐blind fashion. On each study day, renal clearance studies were performed during a 1 h baseline period and subsequently during the second 1 h infusion period. Lithium clearance (CLLi ) was used to estimate proximal tubular outflow. Results Cardiac output increased with the four highest doses. Mean arterial pressure followed a biphasic pattern with a decrease during the two lowest doses and a dose‐dependent increase from the 7.5 μg kg−1 min−1 dose onwards. Effective renal plasma flow increased with all doses of dopamine, but peaked with the 3 μg kg−1 min−1 infusion rate [ from 617 (585–649) ml min−1 with placebo to 915 (824–1006) ml min−1 (means with 95% CI, P&lt;0.001)]. None of the doses changed glomerular filtration rate (GFR). Sodium clearance (CLNa ) and CLLi were elevated with the four lowest doses but increased further from 7.5 μg kg−1 min−1 onwards. Compared with placebo, the percentage increase in CLNa with increasing dose was 77 (5–159), 93 (13–172), 107 (24–190), 121 (60–181), 253 (65–441), 284 (74–494), and 212 (111–312) %, respectively. There were only small, inconsistent decreases in absolute proximal reabsorption rate (APR=GFR‐CLLi ). Fractional distal reabsorption of sodium (FDRNa=(CLLi‐CLNa )/CLLi ) decreased with all doses, reaching its nadir with 7.5 μg kg−1 min−1 [from 95.9 (94.6–97.2) % with placebo to 91.5 (90.0–93.0) % (P&lt;0.01)] whereafter a flat dose‐response curve was observed. Conclusions In conclusion, the renal vasodilating effect of dopamine was maximal with 3 μg kg−1 min−1. The dose‐dependent attenuation seen with higher doses is consistent with an increased α‐adrenergic stimulation opposing the effect on dopaminergic receptors. The present CLLi studies confirm that dopamine increases proximal tubular outflow. The results suggest that the natriuretic effect of depressor doses of dopamine was primarily caused by attenuation of the increase in distal sodium reabsorption normally seen after an increase in proximal tubular outflow. Pressor doses further increased sodium excretion, indicating the presence of pressure natriuresis at these high doses.

Fish oil and cyclosporin A-induced renal hypoperfusion in kidney-transplanted patients

Dietary supplementation with fish oil has been said to improve renal function in cyclosporin A (CsA)-treated subjects. Renal function and the acute renal haemodynamic and tubular response to an oral CsA-dose (3 mg/kg) were investigated before and after 12 weeks of fish oil supplementation (6 g/day) in 12 low-dose CsA-treated kidney-transplanted patients (s-creatinine, 124 +/- 24 mumol/l, mean +/- SD). After an overnight fast, ten l-h renal clearance periods were performed, two periods before and eight after CsA-ingestion. An additional control clearance study without CsA intake was performed in six subjects. Fish oil did not change baseline values of the effective renal plasma flow (ERPF) or the glomerular filtration rate (GFR). Compared to the control study, CsA decreased GFR and ERPF significantly on average 20 +/- 3% (P < 0.01) and 23 +/- 3% (P < 0.01) respectively, 4-6 h after peak CsA blood concentration with no additional effect of fish oil. CsA also significantly decreased the renal clearance of lithium, used as an index of proximal tubular outflow, with no impact of dietary fish oil. Fish oil supplementation had no effect on baseline renal function or CsA-induced hypoperfusion in stable renal transplant recipients treated with a low maintenance dose of CsA.

Renal hemodynamics, tubular function, and response to low-dose dopamine during acute hypoxia in humans

Renal function was investigated in eight normal subjects before and during infusion of dopamine (3 micrograms.kg-1 x min-1) at sea level (SL) and at high altitude (HA, 4,350 m). Lithium clearance (CLi) was used as an index of proximal tubular outflow. HA significantly increased arterial pressure, heart rate, and plasma norepinephrine. Effective renal plasma flow (ERPF) decreased at HA by 10% (P < 0.05), but glomerular filtration rate (GFR), CLi, sodium clearance (CNa), and urine flow remained unchanged compared with SL. Dopamine at SL and HA increased ERPF by 47% (P < 0.001) and 30% (P < 0.01), respectively, but the increase at HA was smaller than that at SL (P < 0.05). Dopamine increased GFR only at SL. CLi and CNa increased by 29% (P < 0.001) and 108% (P < 0.001) at SL and by 23% (P < 0.01) and 108% (P < 0.001) at HA. Whereas dopamine at SL increased urine flow by 46% (P < 0.01), this response was abolished at HA, and free water clearance decreased (P < 0.05). The decreased ERPF at HA suggests a constriction of the renal arterioles secondary to increased adrenergic nervous activity. Although the effect of dopamine on ERPF was attenuated in hypoxia, dopamine-induced increases in CLi and CNa remained unaltered, suggesting that natriuresis in both environments was secondary to an increased outflow from the proximal tubules. The absence of a diuretic response to dopamine at HA seemed to be caused by an effect on distal tubular function.

Effects of acute hypoxia on renal and endocrine function at rest and during graded exercise in hydrated subjects.

Renal effects of altitude hypoxia are unclear. Renal and hormonal function was investigated in eight males at rest and during graded exercise at sea level (SL) and 48 h after rapid ascent to 4,350 m (HA). HA did not change resting values of effective renal plasma flow (ERPF), glomerular filtration rate (GFR), sodium clearance (CNa), urine flow, or lithium clearance (CLi), which was used as an index of proximal tubular outflow. At rest, HA increased plasma norepinephrine concentration and decreased plasma concentrations of renin and aldosterone. Exercise decreased ERPF similarly in both environments. Normoxic exercise progressively reduced GFR, but at HA GFR only decreased during heavy exercise. This resulted in a higher filtration fraction during light and moderate hypoxic exercise. However, calculated absolute proximal reabsorption rate (GFR-CLi) at HA was higher during low-intensity exercise, and there were no significant differences between exercise-induced decreases in CLi, CNa, and urine flow at HA compared with SL. Exercise gradually increased plasma norepinephrine, but values were higher at HA during light and moderate exercise. The small changes in the renal response to low-intensity hypoxic exercise may be secondary to increased adrenosympathetic activity. However, antidiuretic and antinatriuretic effects of exercise were maintained in hypoxia and in both environments seemed to be the consequence of decreased proximal tubular outflow. The results demonstrate that renal glomerular and tubular function is well preserved in acute hypoxia despite marked hormonal changes.

Prazosin attenuates the natriuretic response to atrial natriuretic factor in man

The effect of alpha-1-adrenoceptor blockade with 0.25 mg oral prazosin on the renal response to atrial natriuretic factor (ANF) 5 pmol/kg/min was examined in eight healthy male volunteers undergoing maximal water diuresis. ANF on its own decreased mean arterial blood pressure (P less than 0.05) without altering heart rate or increasing plasma norepinephrine. ANF increased urinary sodium excretion by 130% (P less than 0.01) from baseline value with accompanying 18% decrease (P less than 0.05) in PAH clearance (ERPF) without changing inulin clearance (GFR). When compared to placebo infusion, ANF infusion caused a significant increase in fractional excretion lithium (FELi), a marker of proximal tubular function. Fractional distal delivery of sodium, another marker of proximal tubular outflow as determined by free water clearance, was also increased during ANF infusion. As expected, ANF decreased distal nephron fractional sodium reabsorption as evaluated by both the "lithium method" and by the conventional "solute-free water method." Prazosin on its own had no effect on blood pressure, renal function or hormonal parameters. When given in combination with ANF, prazosin blunted the natriuretic effect of ANF from 130% to 35% (P less than 0.01). However, prazosin pretreatment did not influence the ANF-induced fall in blood pressure or ERPF nor the ANF-induced suppression of plasma aldosterone. We have therefore found evidence to support the hypothesis that at basal levels of sympathetic tone, the natriuretic effect of ANF in man is dependent on an intact sympathetic nervous system, since sympathetic blockade by prazosin blunts its sodium excretory effects.

Renal tubular reabsorption of sodium and water during infusion of low-dose dopamine in normal man

1. Using the renal clearance of lithium (CLi) as an index of proximal tubular outflow of sodium and water, together with simultaneous measurements of effective renal plasma flow, glomerular filtration rate (GFR) and sodium clearance (CNa), renal function and the tubular segmental reabsorption rates of sodium and water during dopamine infusion (3 micrograms min-1 kg-1) were estimated in 12 normal volunteers. 2. CNa increased by 128% (P less than 0.001). Effective renal plasma flow and GFR increased by 43% (P less than 0.001) and 9% (P less than 0.01), respectively. CLi increased in all subjects by, on average, 44% (P less than 0.001). Fractional proximal reabsorption [1-(CLi/GFR)] decreased by 13% after dopamine infusion (P less than 0.001), and estimated absolute proximal reabsorption rate (GFR-CLi) decreased by 8% (P less than 0.01). Absolute distal sodium reabsorption rate [(CLi-CNa) x PNa, where PNa is plasma sodium concentration] increased (P less than 0.001), and fractional distal sodium reabsorption [(CLi-CNa)/CLi] decreased (P less than 0.001). 3. It is concluded that natriuresis during low-dose dopamine infusion is caused by an increased outflow of sodium from the proximal tubules that is not fully compensated for in the distal tubules.

Overall renal and tubular function during infusion of amino acids in normal man

1. Amino acids have been used to test renal reserve filtration capacity. Previous studies suggest that amino acids increase glomerular filtration rate (GFR) by reducing distal tubular flow and tubuloglomerular feedback activity. 2. Glomerular function and the renal tubular handling of sodium during infusion of amino acids was studied in 12 normal volunteers. 3. Clearance of sodium (CNa) was unchanged. Effective renal plasma flow increased slightly, but significantly, by 9% (P less than 0.05). GFR was increased by 13% (P less than 0.001). Clearance of lithium (CLi) (used as an index of proximal tubular outflow) increased by 38% (P less than 0.001). Calculated absolute proximal reabsorption (GFR-CLi) remained unchanged. Fractional proximal reabsorption [1-(CLi/GFR)] was decreased by 10% (P less than 0.001). Calculated absolute distal sodium reabsorption [(CLi-CNa) x PNa, where PNa is plasma sodium concentration] increased by 40% (P less than 0.001). Plasma renin concentration did not change significantly. 4. The results suggest that amino acids increase GFR by a primary effect on renal haemodynamics or, less likely, by reducing the signal to the tubuloglomerular feedback mechanism. The increase in proximal tubular outflow was compensated for in the distal tubules, so that the sodium excretion rate remained unchanged.

Evidence for a Proximal and Distal Nephron Action of Atrial Natriuretic Factor in Man

The intrarenal mechanism of action of atrial natriuretic (ANF) remains uncertain. Animal work suggests that part of the natriuretic effect of ANF may be due to inhibition of proximal and distal nephron sodium reabsorption and we now present evidence for similar effects of ANF in man. Six sodium replete normal male volunteers were studied during maximal water diuresis. Lithium clearance was used to assess segmental nephron function. ANF infusion caused a significant increase in fractional lithium clearance (FELi) compared to placebo infusion. A similar change in fractional distal delivery, a conventional marker of proximal tubular outflow, also occurred during ANF infusion independently corroborating the increase in FELi. These findings suggest that ANF inhibits whole-kidney fractional proximal tubular sodium reabsorption in man. Evidence is also presented to show that ANF depresses distal nephron fractional sodium reabsorption as evaluated both by the lithium method and by estimation of solute-free water clearance.