bluuebunny/crossref_metadata_2025 huggingface dataset:
Aldosterone and corticosterone binding and effects on Na+ transport in cultured kidney cells
Authors: Watlington C. O. , Perkins F. M. , Munson P. J. , Handler J. S.
Published: December 2017

A continuous line of cells (A6) derived from toad kidney has been shown to form epithelia in culture that manifest aldosterone-stimulatable transepithelial sodium transport. In this study an efficient filtration assay for nuclear binding of [3H]aldosterone was validated. Specific high-affinity aldosterone and corticosterone binding sites in the particulate (nuclear-enriched) fraction were characterized in intact epithelia. Despite metabolism of both steroids, two high-affinity binding sites for each were demonstrable: aldosterone, K'd1 = 0.85 (+/- 0.19) X 10(-10) and K'd2 = 1.6 (+/- 0.42) X 10(-8) M; corticosterone, K'd1 = 0.5 (+/- 0.31) X 10(-10) and K'd2 = 0.32 (+/- 0.19) X 10(-8) M. Analogue competition-binding studies indicated a qualitative difference in the two sites and co-occupancy of both sites by the two steroids. The sodium transport response to aldosterone and corticosterone approximated a linear function of occupancy of the lower affinity sites. Although the lower affinity sites resemble mammalian glucocorticoid receptors in terms of relative binding affinities for analogues, we conclude that they are the receptors which mediate the aldosterone and corticosterone stimulation of Na+ transport in these epithelia.
Regulation of extrarenal potassium homeostasis by adrenal hormones in rats
Authors: Bia M. J. , Tyler K. A. , DeFronzo R. A.
Published: December 2017

The effect of chronic (7-10 days) adrenal insufficiency on extrarenal potassium tolerance was examined by infusing potassium into rats after acute nephrectomy. The increment in plasma potassium concentration was significantly higher in glucocorticoid-replaced adrenalectomized rats versus controls (max delta PK 3.59 +/-0.11 vs. 2.93 +/- 0.08 meq/liter; P less than 0.001). The impairment in extrarenal potassium tolerance in adrenalectomized rats could not be attributed to acidemia, hypotension, changes in plasma insulin or glucose concentration, or potassium retention prior to study. Acute replacement with aldosterone resulted in significant improvement in the rise in plasma potassium after KCl (max delta PK 3.18 +/- 0.06 meq/liter; P less than 0.005 compared with aldosterone-deficient adrenalectomized rats but higher than in controls, P less than 0.02). If given on a chronic basis, aldosterone replacement led to a complete correction of the defect (max delta PK = 2.89 +/- 0.08 meq/liter). Acute epinephrine replacement in adrenalectomized rats also returned potassium tolerance to normal (max delta PK = 3.02 +/- 0.10 meq/liter). The results demonstrate that extrarenal potassium tolerance is impaired in chronic adrenal insufficiency and suggest that both aldosterone and epinephrine deficiency may contribute to the defect, since replacement with either hormone returns potassium tolerance toward normal. Accordingly, both aldosterone and epinephrine have important extrarenal mechanisms of action.
Uptake of Pi in brush border vesicles after release of unilateral ureteral obstruction
Authors: Weinreb S. , Hruska K. A. , Klahr S. , Hammerman M. R.
Published: December 2017

After release of complete unilateral ureteral obstruction, a decreased fractional excretion of phosphate (Pi) is observed in the postobstructed kidney compared with the nonobstructed (control) kidney. To determine whether this decrease in the urinary excretion of Pi is due to changes in Na+-dependent Pi transport across the renal brush border membranes of postobstructed and control kidneys, membrane vesicles were prepared from the brush borders of kidneys from dogs that had undergone complete unilateral ureteral obstruction. Alkaline phosphatase activity was decreased in membrane vesicles isolated from postobstructed kidneys. No differences were observed in Na+-dependent Pi transport or in Na+ uptake in membrane vesicles isolated from postobstructed as compared with control kidneys. The in vivo administration of parathyroid hormone decreased Na+-dependent Pi transport in membrane vesicles isolated from postobstructed and control kidneys despite the absence of a phosphaturic response. Our findings suggest that no intrinsic change in the transport characteristics of Pi across the luminal membrane of renal tubular cells occurs with unilateral ureteral obstruction. The findings are consistent with the suggestion that the low fractional excretion of Pi in the postobstructed kidney results from very low filtered loads of Pi on the postobstructed side.
Water metabolism after cisplatin in the rat
Authors: Gordon J. A. , Peterson L. N. , Anderson R. J.
Published: December 2017

A polyuric state is often observed after cis-dichlorodiammine platinum (cisplatin). To study the mechanism of this polyuria we gave 5-6 mg/kg cisplatin to conscious rats and observed polydipsia and a polyuric form of mild acute renal failure. A defect in renal concentrating ability was observed 1 and 8 days after cisplatin. Animals demonstrated diminished postdehydration plasma vasopressin at 1 but not at 8 days after cisplatin, and exogenous vasopressin corrected the renal concentration defect at 1 but not at 8 days after cisplatin. To assess the role of polydipsia in the concentration defect, water intake in cisplatin-treated animals was matched to pair-fed controls. Prevention of polydipsia improved the polyuria but not the concentration defect seen 8 days after cisplatin. To assess intrarenal factors in the renal concentration defect, postdehydration interstitial solute was measured and was significantly lower in cisplatin-treated than in control animals. To determine whether the diminished interstitial solute was due to vascular mechanisms, inner medullary plasma flow was measured and was identical in cisplatin-treated and control rats. Treatment with cisplatin also resulted in decreased excretion of a water load. We conclude that either impaired synthesis or release of vasopressin is the cause of the impaired renal concentration seen 1 day after cisplatin. Eight days after cisplatin, the renal concentration defect is due in part to decreased interstitial tonicity.
Bicarbonate reabsorption and potassium handling in isolated perfused kidneys of Bufo marinus
Authors: Long S.
Published: December 2017

Average rates of bicarbonate reabsorption, normalized for glomerular filtration rates, rose linearly with perfusate [HCO3-] of 6-21 mM with a constant bicarbonate/creatinine clearance ratio of 0.06, similar to the in vivo value. Maximal rates of bicarbonate reabsorption occurred at 20-25 mM HCO3- perfusate. At 3 mM perfusate [K+], normalized rates of potassium reabsorption were constant from filtered perfusates mimicking metabolic acidosis, and from control perfusates; during perfusion with alkalotic Ringer solutions net potassium secretion was observed. Elevation of perfusate [K+] to 10 mM in alkalotic Ringer significantly decreased maximal rates of bicarbonate reabsorption. Overall, the pattern of bicarbonate reabsorption in toad kidneys perfused at 6-21 mM HCO3- was similar to that observed in acid-base disturbances of metabolic origin in mammals, while the pattern of potassium excretion showed significant differences in these two vertebrate classes.
Fate of [125I]insulin removed from the peritubular circulation of isolated perfused rat kidney
Authors: Petersen J. , Kitaji J. , Duckworth W. C. , Rabkin R.
Published: December 2017

Although there is considerable evidence that insulin is removed from the peritubular circulation of the mammalian kidney, it is unclear whether binding to insulin-specific receptors is involved in this process, whether after peritubular removal the hormone is degraded to small fragments with release into the circulation, or whether it merely undergoes a minor modification with loss of immunoreactivity. We examined the metabolism of [125I]insulin removed from the peritubular circulation of the nonfiltering isolated perfused rat kidney and compared it to that of [125I]insulin metabolized by filtering isolated kidneys and kidney homogenates. The results indicate that after peritubular removal, a small amount of insulin is degraded to form low-molecular-weight products similar to those seen with filtering kidneys and kidney homogenates. However, most of the insulin removed from the peritubular circulation is processed either to nonimmunoreactive products of molecular weight similar to that of insulin or, to a lesser extent, to products of larger molecular weight. Both these products are also formed by filtering kidneys. In the filtering kidney, the products having molecular weight similar to that of insulin probably originate from the peritubular process, because it is unlikely that material of this size could be derived from the filtration-absorption pathway. Of particular note was the finding that [125I]insulin trapped in the peritubular compartment of nonfiltering kidneys was displaced severalfold more effectively by unlabeled insulin than by several peptide hormones (P less than 0.01); the latter were no more effective than vehicle alone. The findings suggest the presence of peritubular insulin-specific receptors.
Effects of calcium on ADH action in the cortical collecting tubule perfused in vitro
Authors: Goldfarb S.
Published: December 2017

To test the effects of calcium on ADH action in an in vitro mammalian system, the rabbit cortical collecting tubule was studied. After 25 microunits/ml ADH (n=8) in the presence of 1.25 mM calcium bath, water flow (Jv) rose to 1.56 +/- 0.34 nl.mm-1. min-1 and hydraulic conductivity (Lp, cm.s-1.atm-1 X 10(7)) rose to 123 +/- 22. After 25 microunits/ml ADH in the presence of 3.75 mM calcium bath (n=7), Jv rose to 2.96 +/- 0.6 nl.mm-1.min-1 (P less than 0.05 vs. control) and Lp rose to 286 +/- 62 cm.s-1.atm-1 X 10(7) (P less than 0.02 vs. 1.25 mM bath calcium control). Tubules (n=6) perfused with 3.75 mM Ca and bathed in 3.75 mM Ca also showed an Lp of 279 +/- 82 cm.s-1.atm-1 X 10 (7) following 25 microunits/ml ADH. Tubules similarly studied in 1.25 (n=6) or 3.75 mM Ca (n=6) bath but treated with 10(-4) M 8-[p-chlorophenylthio]cAMP demonstrated Lp of 222 +/- 26 and 235 +/- 37 cm.s-1.atm-1 X 10(7), respectively. These data suggest that increased bath Ca enhances ADH- but not cAMP-stimulated water flow in the rabbit cortical collecting tubule. High perfusate Ca2+ does not alter the stimulatory effect of elevated peritubular Ca2+.
Hydroosmotic response of collecting tubules to ADH or cAMP at reduced peritubular sodium
Authors: Frindt G. , Windhager E. E. , Taylor A.
Published: December 2017

Changes in cytosolic Ca2+ activity have been implicated in the hydroosmotic response to vasopressin (VP) in amphibian urinary bladder; the level of cytosolic free Ca2+ may be regulated, in part, by a process of Na-Ca exchange across the basolateral cell membrane. To assess whether similar mechanisms operate in the mammalian nephron, the effect of low peritubular [NA] on the hydraulic conductivity (Lp) of the isolated perfused rabbit collecting tubule (CT) exposed to either 20 microunits/ml VP or 5 X 10(-5) M 8-[p-chlorophenylthio]-cyclic 3',5'-adenosine monophosphate (ClPheS-cAMP) was studied. Low peritubular [Na] had no effect on the basal water permeability of the CT. After exposure to VP, CTs bathed in 145 mM Na Ringer solution developed an Lp of 324 +/- 27 X 10(-7) cm.s-1.atm-1, while tubules bathed in 4 mM Na and 141 mM tetramethylammonium Ringer solution achieved an Lp of only 112 +/- 13 X 10(-7) cm.s-1.atm-1 (P less than 0.001). Inhibition of the VP response was estimated to be half-maximal when peritubular Na was 120 mM. The hydroosmotic response to ClPheS-cAMP was diminished by 44% of the control values when CTs were bathed in a 4 mM Na medium; this inhibition was greatly attenuated when the peritubular Ca concentration was reduced to 0.05 mM. These results are consistent with the view that 1) a Na-Ca exchange process operates at the basolateral surface of the mammalian cortical collecting tubule cells, and 2) elevated cytosolic Ca ion activity inhibits the increase in water permeability elicited by VP or cAMP in this nephron segment.
Effect of furosemide on the electrical parameters of frog skin
Authors: Corcia A. , Caplan S. R.
Published: December 2017

When added to the mucosal solution bathing isolated frog skin at concentrations ranging from 5 X 10(-4) to 3 X 10(-3) M, the diuretic furosemide increased both the active transport of sodium and the electrical potential difference across the tissue in a dose-dependent way. The same effect was observed in chloride-free solutions. Mucosal furosemide also decreased the passive unidirectional fluxes of chloride. We believe that as far as electrical parameters are concerned mucosal furosemide has a double effect in frog skin: it increases the active conductance to sodium across the mucosal membrane, thus increasing active transport, and decreases the passive permeability to chloride, thus altering the passive conductance of the skin. The relative increase in short-circuit current was, however, invariably greater than the increase of the active conductance, suggesting the influence of yet a third effect. The effect of mucosal furosemide on active sodium transport was blocked by amiloride (5 X 1-(-5) M) and was independent of vasopressin. Qualitatively the effect was similar to the effect produced by triphenylmethylphosphonium ion.
Effect of furosemide on the thermodynamic parameters of frog skin
Authors: Corcia A. , Caplan S. R.
Published: December 2017

The formalism of nonequilibrium thermodynamics (NET) was used to analyze the effect of diuretic furosemide on the transport mechanism of frog skin. Mucosal furosemide increased the active conductance of Na+ across the mucosal membrane of the cells, producing an increased transport of Na+ (short-circuit current). Furosemide also increased both the thermodynamic affinity of the metabolic reaction supplying energy to the sodium pump and the degree of coupling between transport and metabolism. It changed the phenomenological cross-coefficient of the NET description as well as the stoichiometric ratio, indicating that its effect cannot be explained simply on the basis of a change in Na+ conductance. The effect on the NET parameters was independent of the presence of either chloride or sulfate as the principal anion in the solutions, and was qualitatively similar to the effect produced by mucosal application of triphenylmethylphosphonium ion.
Effect of respiratory acidosis on acidification by the medullary collecting duct
Authors: Bengele H. H. , Graber M. L. , Alexander E. A.
Published: December 2017

The effect of acute respiratory acidosis (ARA) on inner medullary collecting duct (IMCD) acidification was studied and the results were compared with previously obtained data by our laboratory in rats with acute metabolic acidosis (AMA). We employed the microcatheterization technique to directly measure pH and PCO2 with glass-membrane electrodes, and fluid samples were obtained for measurement of bicarbonate, phosphate, and ammonium. Arterial pH was 7.18 +/- 0.01 and PCO2 was 88 +/- 2 mmHg. The IMCD data were analyzed as a function of IMCD length (approximately 6 mm). pH decreased from 5.78 +/- 0.07 to 5.27 +/- 0.03 and PCO2 increased from 55 +/- 4 to 75 +/- 2 mmHg between origin and tip. Bicarbonate delivery decreased from 154 +/- 34 to 25 +/- 3 nmol/min but no change was noted in acid phosphate, ammonium, or net acid addition along the IMCD. However, net acid excretion was not different from that found previously in AMA. We conclude that during ARA acidification is augmented prior to, but not along, the IMCD. In contrast, during AMA we previously found that IMCD plays a major regulatory role in urinary acidification, accounting for about 50% of the excreted hydrogen ion.
Vanadium, Na-K-ATPase, and potassium adaptation in the rat
Authors: Higashino H. , Bogden J. D. , Lavenhar M. A. , Bauman J. W. , Hirotsu T. , Aviv A.
Published: December 2017

Vanadate is a potent inhibitor of Na-K-ATPase in vitro. It has been suggested that vanadium may function as a cellular regulator of Na-K-ATPase in vivo. To examine this speculation, we studied in rats the effect of high vanadate intake on 1) the tissue levels and distribution of vanadium, 2) basal activity of Na-K-ATPase in various tissues, and 3) the activity of Na-K-ATPase in various organs under conditions of massive chronic potassium loading known to stimulate Na-K-ATPase in the kidney and colon. Despite extremely high tissue levels of vanadium there was no demonstrable effect of the element on the basal activity of Na-K-ATPase. When subjected to chronic potassium loading, rats with high tissue vanadium concentrations underwent potassium adaptation that was associated with a rise in Na-K-ATPase activity in the renal cortex, renal medulla, and colonic mucosa. Further studies are needed to support or refute the thesis that vanadium might be an intracellular regulator of Na-K-ATPase in vivo.
Hormonal modulation of glomerular function
Authors: Dworkin L. D. , Ichikawa I. , Brenner B. M.
Published: December 2017

Glomeruli contain receptors for many hormones. Binding of angiotensin II (ANG II) or antidiuretic hormone (ADH) to glomerular mesangial cells elicits a contractile response. Other hormones induce synthesis of cyclic nucleotides (cAMP, cGMP). Glomeruli also synthesize several prostaglandins, renin, and ANG II. Micropuncture studies in Munich-Wistar rats have examined the effects of vasoactive drugs and hormones on the filtration process. Several vasodilators increase renal plasma flow in the dog and rat, but GFR remains relatively unchanged due to an offsetting fall in the ultrafiltration coefficient (Kf). Vasoconstrictor substances such as ANG II and norepinephrine cause declines in renal plasma flow and Kf, but GFR remains constant due to an increase in the transcapillary hydraulic pressure gradient. Antidiuretic peptides and parathyroid hormone also reduce Kf. Glomerular mesangial cells may regulate Kf by contracting and reducing glomerular capillary surface area. ANG II and ADH directly stimulate mesangial cell contraction in vitro. Other hormones appear to cause contraction by inducing local ANG II synthesis. These hormonal pathways are implicated in the pathogenesis of altered glomerular function in diverse forms of renal injury.
Liquid junctions and isolated proximal tubule transepithelial potentials
Authors: Laprade R. , Cardinal J.
Published: December 2017

Accurate evaluation of the unavoidable liquid junction potentials (LJP) involved whenever bath (B) and perfusate (P) solutions differ in ionic composition is of fundamental importance in the determination of proximal convoluted tubule (PCT) transepithelial potential, psi T, and its active and diffusion components, psi A and psi D. Therefore, a precise method of measuring these LJP, using Ag/AgCl electrodes, has been developed and successfully tested. The measured LJP, psi B-P, between artificial bath serum containing 6 g/100 ml albumin and ultrafiltrate-like perfusate is +2.6 mV. The average measured potential difference (PD) in rabbit PCT with these solutions was -3.0 mV, psi T thus being only -0.4 mV. Three maneuvers known to abolish psi A (hypothermia, 10(-4) ouabain, or K-free bath) brought the PD close to the reference potential, indicating that psi A congruent to PD and that psi D congruent to psi B-P. From dilution potentials, after correction for LJP (4.3 in the absence and 6.2 mV in the presence of proteins), PNa/PCl was calculated to be 0.9 for midcortical and 1.1 for juxtamedullary PCT in the absence of proteins and slightly lower in their presence. These values of LJP obtained with Ag/AgCl electrodes are between 30 and 100% larger than the ones measured using a saturated KCl bridge or the ones in previous studies, indicating that in these studies psi T as well as PNa/PCl has been overestimated.
Renal tubular uptake of protein: effect of molecular charge
Authors: Christensen E. I. , Rennke H. G. , Carone F. A.
Published: December 2017

The effect of molecular charge of proteins on proximal tubular reabsorption was evaluated in the rat. Native and two cationized forms of albumin, native and anionized lysozyme, and native and anionized cytochrome c were iodinated with 125I. The different forms of each type of protein were alternately microinfused into the same site of proximal convoluted tubules in vivo. Tubular reabsorption was determined as the difference between the amounts of TCA-precipitable radioactivity infused and recovered in the urine. At low concentration of albumin 5 times more cationized than anionic albumin and 2.7 times more cationic than anionized lysozyme were reabsorbed by the proximal tubule. At two of four concentrations, proximal tubular uptake of cationic cytochrome c exceeded that of anionized cytochrome c. Uptake of cationic cytochrome c exceeded that of cationic lysozyme; however, the difference in uptake between native cationic and anionized species of the two proteins was much greater for lysozyme than for cytochrome c. The data reveal that a higher isoelectric point significantly enhances proximal tubular reabsorption of albumin, lysozyme, and cytochrome c and that proteins with similar molecular weight and isoelectric point are not necessarily reabsorbed to the same degree. This suggests that in addition to total molecular charge the molecular configuration and/or distribution of electrical charges on teh protein surface determine protein binding by the luminal membrane and subsequent endocytosis by the proximal tubule.
Autoregulation of renal blood flow in the rat: effects of growth and uninephrectomy
Authors: Chevalier R. L. , Kaiser D. L.
Published: December 2017

As a result of normal maturation or after reduction in renal mass, renal blood flow (RBF) progressively increases. However, the effects of renal growth on the relationship of RBF to renal perfusion pressure (RPP) have not been systematically investigated. We examined RBF as a function of RPP in anesthetized young and adult rats that had been subjected to uninephrectomy or sham operation 3-4 wk previously. As a result of normal growth, mean arterial blood pressure increased from 94.1 +/- 2.7 (SE) to 118.9 +/- 4.2 mmHg. The calculated autoregulation factor, in which a value less than 1 indicates the presence of autoregulation, was 0.44 +/- 0.10 over RPP 70-100 mmHg and 1.74 +/- 0.25 over RPP 40-70 mmHg in sham-operated young animals. In adult sham-operated rats, the factor was 0.38 +/- 0.07 over RPP 100-130 mmHg and 1.03 +/- 0.07 over RPP 70-100 mmHg. Uninephrectomy in adults resulted in a 30% rise in RBF over RPP 100-130 mmHg, and there was no change in the autoregulation factor. Uninephrectomy in young rats, however, resulted in a 35% rise in RBF at RPP = 100 mmHg with only a 17% rise at RPP = 70 mmHg, and the autoregulation factor increased to 0.91 +/- 0.10 over this range of RPP. We conclude that the autoregulatory range shifts with increasing blood pressure during normal growth and that autoregulation is "reset" to higher RBF in the uninephrectomized adult rat. Although autoregulation takes place in the young animal, uninephrectomy alters the relationship of RBF to RPP such that autoregulation is impaired.
Organization of nephron function
Authors: Knepper M. , Burg M.
Published: December 2017

Recent studies of mammalian nephron segments have revealed an unexpected diversity of renal transport functions. Most substances are transported by several segments, and the transport mechanisms differ from segment to segment. In this paper we review some of these findings in order to fit them into an integrated picture of kidney function. The main question we ask is what is the advantage of spatial separation of the various transporters along the nephron. We propose that spatial separation of transport functions allows independent and efficient control of the excretion of substances whose renal handling is interdependent. The organization of sodium and water handling along the nephron is considered in the greatest detail. Sodium and water play central roles in controlling the renal excretion of many other substances. Yet the excretion rates of sodium and water themselves are independently controlled. The intricate anatomical structure of the kidney not only provides spatial separation of transport processes along the nephron but also couples the function of different segments by juxtaposing them in specialized regions within the medulla and cortex. These anatomical arrangements provide a framework for integrating the complex array of renal functions.
Phosphate uptake by renal membrane vesicles of rabbits adapted to high and low phosphorus diets
Authors: Cheng L. , Liang C. T. , Sacktor B.
Published: December 2017

Renal adaptation to changes in phosphate intake was studied by comparing phosphate uptake by proximal tubule brush border membrane vesicles from rabbits on a relatively high or low phosphorus diet. The low phosphorus diet increased Na+ gradient-dependent phosphate uptake. Uptake in the absence of Na+ and in the presence of Na+, but no gradient, was not significantly affected. The phosphorus diet did not alter Na+ gradient-dependent D-glucose and L-proline uptake. The low phosphorus diet increased Vmax; affinity for phosphate was not appreciably changed. At all concentrations of extravesicular Na+, phosphate uptake was higher in membrane vesicles from animals fed the low phosphorus diet; the kinetics of the phosphate uptake system, with respect to Na+, was also altered by the change in dietary phosphate. These findings suggest that adaptation involves an alteration in the rate of translocation of the Na+-phosphate carrier when energized by a Na+ gradient driving force rather than a change in the number of Na+-phosphate carrier sites. With membrane vesicles from rabbits fed a low phosphorus diet, phosphate uptake increased several-fold when the pH of the uptake medium was raised, whereas with membrane vesicles from animals fed a high phosphorus diet the enhancement of uptake with alkalinization was relatively small. Irrespective of the diet, divalent phosphate was the probable preferred species for transport. Dietary adaptation was associated, however, with an alteration in the pH dependency of the transport system per se. These findings provide evidence that the adaptation of the kidney phosphate transport system to dietary phosphate load involves an intrinsic change in the Na+-phosphate carrier.
Inhibitory effect of epinephrine on renal potassium secretion: a micropuncture study
Authors: DeFronzo R. A. , Stanton B. , Klein-Robbenhaar G. , Giebisch G.
Published: December 2017

The effect of epinephrine on renal potassium excretion was examined in the rat. In group I KCl was infused acutely to increase plasma K (PK) by 2.0 meq/liter; urinary K excretion (UKV) rose by 1.22 mueq/min. In group II rats, which received a similar dose of KCl but with epinephrine, the increase in PK (delta = 0.8 meq/liter, P less than 0.001) was blunted and UKV was reduced (delta = 0.23 mueq/min, P less than 0.001). To determine whether the reduction in UKV resulted from the smaller increase in PK or from a direct action of epinephrine on renal K transport, a third group of animals received a lower dose of KCl. Despite similar PK levels, the epinephrine group excreted significantly less K in the urine (0.61 vs. 0.93 mueq/min). In group IV propranolol was infused with KCl; UKV was modestly increased. The effects of epinephrine on UKV were unrelated to changes in glomerular filtration rate, urine flow, or UNaV. Micropuncture results showed that at comparable PK levels epinephrine had no direct effect on K secretion by the distal tubule but indirectly inhibited K secretion in this nephron segment by reducing PK. In addition, epinephrine reduced K addition at tubular sites beyond the late distal tubule, most likely in the collecting tubule.
Renal tubular calcium transport: effects of changes in filtered calcium load
Authors: Sutton R. A. , Wong N. L. , Quamme G. A. , Dirks J. H.
Published: December 2017

Micropuncture experiments were performed in thyroparathyroidectomized dogs to examine the influence of changes in filtered calcium load on segmental tubular calcium reabsorption. Filtered calcium load was changed either by reducing glomerular filtration rate (GFR) by aortic clamping (group I) or by progressive calcium infusion (group II) to increase plasma ultrafilterable calcium concentration (UFCa). The results suggest that fractional proximal calcium reabsorption responds similarly to altered filtered load, whether produced by changes in GFR or UFCa. In contrast, fractional reabsorption by the loop segment is progressively reduced as UFCa is increased but is relatively unchanged by alterations in filtered load secondary to altered GFR. These data indicate a specific parathyroid hormone-independent reciprocal effect of UFCa on calcium reabsorption in the loop segment, which may be an important determinant of urinary calcium excretion.
Chloride dependence of the HCO3 exit step in urinary acidification by the turtle bladder
Authors: Fischer J. L. , Husted R. F. , Steinmetz P. R.
Published: December 2017

To characterize the efflux of HCO-3 across the basolateral membrane of the H+-secreting cells of the turtle bladder, we examined the effect of substitution of gluconate or methyl sulfate for Cl- on the rate of acidification (JH). JH was measured as the short-circuit current in bladders in which Na+ transport was abolished with 10(-4) M ouabain. In hemibladders bathed in normal Ringer solution (Cl- = 122 mM) JH was 44.9 microA. Substitution of the Cl- resulted in a marked reduction in JH (12.5 microA with gluconate and 7.5 microA with methyl sulfate). Addition of Cl- to the mucosal surface had no effect on JH. In contrast, serosal addition of Cl- restored JH to control. The apparent Km for Cl- in gluconate Ringer was 0.13 mM. Serosal furosemide (1 mM) inhibited JH by 55% in Cl- Ringer. We conclude that HCO-3 exit across the basolateral membrane of the H+-secreting cell occurs via a Cl-HCO3 exchanger that has a high affinity for chloride.
Passive driving forces of proximal tubular fluid and bicarbonate transport: gradient dependence of H+ secretion
Authors: Chan Y. L. , Malnic G. , Giebisch G.
Published: December 2017

The effect of oncotic pressure changes on fluid (Jv) and net bicarbonate transport (JHCO-3) and the transepithelial bicarbonate permeability (PHCO-3) were measured by an improved luminal and capillary microperfusion method that allows paired experiments on the same tubule. Rat proximal tubules were pump-perfused and Jv and [HCO-3] measured with [14C]inulin and a pH glass electrode. Raising peritubular protein (0-8-15 g/100 ml bovine serum albumin) stimulated Jv and HCO-3 reabsorption. The response to oncotic pressure changes was asymmetrical since changes of the luminal protein concentration had no significant effects. Whereas transepithelial solvent drag effects on HCO-3 must be minimal, peritubular protein most likely stimulates translocation of fluid and bicarbonate from intercellular spaces into peritubular capillaries. PHCO-3 was measured from HCO-3 net flux along a lumen-to-capillary-directed electrochemical potential gradient. In these experiments active H+ transport and Jv were minimized by 10(-4) M acetazolamide and luminal raffinose. PHCO-3 was 1.77 X 10(-5) cm X s-1 and was unaffected by increasing luminal flow rate from 10 to 45 nl X min-1. Since bicarbonate backflux is only a small fraction of physiological rates of JHCO-3, net transport alterations at varying [HCO-3] in the lumen must be due to changes in active HCO-3 (H+) transport. Thus, active H+ ion secretion across the luminal membrane of the proximal tubule is gradient dependent.
Inhibition of vasopressin action by vanadate in the cortical collecting tubule
Authors: Edwards R. M. , Grantham J. J.
Published: December 2017

The effect of vanadate, a potent inhibitor of Na-K-ATPase, on the hydroosmotic response to vasopressin (AVP) and transepithelial voltage (Vt) in cortical collecting tubules was examined. At 37 degrees C, exposure of collecting tubules to bath vanadate (10(-4) M) for 30 min inhibited the increase in hydraulic water permeability (Lp) in response to AVP or 8-bromo-cyclic adenosine monophosphate by 68 and 76%, respectively. When vanadate was present only in the lumen no inhibition of the AVP response was observed. Incubation of tubules with ouabain (10(-5) M) for 30 min inhibited the AVP-induced increase in Lp to the same extent as vanadate. At 25 degrees C, vanadate inhibited the increase in Lp by AVP if added before but not after the hormone. Addition of vanadate to the bath caused a rapid decrease in the lumen-negative Vt that is consistent with Na-K-ATPase inhibition. Luminal vanadate also inhibited Vt but the rate of decrease of Vt was much slower than in the presence of bath vanadate. We conclude that vanadate inhibits the development but not the maintenance of the AVP-induced increase in water permeability in the collecting tubule. Since the effect of ouabain was similar to that of vanadate, the results suggest that inhibition of Na-K-ATPase directly or indirectly interferes with the initiation of the AVP-induced increase in luminal membrane water permeability at a site distal to cAMP formation.
Altered osmotic thresholds for vasopressin secretion and thirst in human pregnancy
Authors: Davison J. M. , Gilmore E. A. , Durr J. , Robertson G. L. , Lindheimer M. D.
Published: December 2017

Osmoregulation was studied in eight women during late pregnancy and again 8-10 wk postpartum. Base-line plasma osmolality (Posmol) was significantly lower during (280.9 +/- 2.1 mosmol/kg, SD) than after (289.4 +/- 2.1 mosmol/kg) pregnancy yet 24-h urinary volume and plasma arginine vasopressin (PAVP) measured in vasopressinase-inactivated blood was similar in both groups (pregnancy, 1.39 +/- 0.56 pg/ml; postpartum, 1.25 +/- 0.62 pg/ml). After 12 h of dehydration PAVP rose similarly and significantly both during (2.25 +/- 0.81 pg/ml) and after (2.89 +/- 1.19 pg/ml) gestation, and Uosmol was similar on both occasions (pregnancy, 779 +/- 121 mosmol/kg; postpartum, 784 +/- 102 mosmol/kg). When Posmol was increased by the slow infusion of 5% saline PAVP increased as soon as body tonicity did both during and after pregnancy. PAVP correlated significantly with Posmol in each subject (range of r, 0.75-0.99) and the mean regression lines [pregnancy, PAVP = 0.32 (Posmol; -279), r = 0.79; postpartum, PAVP = 0.38 (Posmol, -285), r = 0.86] demonstrated that the apparent osmotic threshold for AVP secretion was 6 mosmol/kg lower during than after gestation. Similarly the Posmol at which the subject experienced a conscious desire to drink was lower in pregnant (287 +/- 1.6 mosmol/kg) compared with postpartum subjects (298 +/- 2.0 mosmol/kg; P less than 0.001). These data demonstrate decreased osmotic thresholds for AVP release and thirst during human pregnancy and explain why gravidas can maintain their new lower Posmol within narrow limits.
Inner medullary hemodynamics in chronic salt-depleted dogs
Authors: Chou S. Y. , Spitalewitz S. , Faubert P. F. , Park I. Y. , Porush J. G.
Published: December 2017

The effects of chronic salt depletion on medullary hemodynamics remain unknown. In the present study, sodium excretion, renal hemodynamics including papillary plasma flow, measured by the albumin-accumulation technique, and papillary tissue solute content were determined during hydropenia in 13 anesthetized sodium-replete and 10 sodium-depleted dogs. Salt depletion induced a significant rise in plasma renin activity and aldosterone without potassium depletion. Mean arterial pressure, GFR, and renal blood flow were similar in sodium-depleted and sodium-replete dogs. Despite a similar distribution of cortical blood flow (measured by the microsphere method) in the two groups, papillary plasma flow was markedly reduced in sodium-depleted dogs (8.8 +/- 1.7 vs. 22.8 +/- 1.9 ml X min-1 X 100 g-1 in sodium-replete dogs), associated with a significant decrease in renal sodium excretion. Furthermore, papillary osmolality and sodium concentration were significantly greater in sodium-depleted dogs. Ultrastructure examination revealed smooth muscle cells surrounding the efferent arterioles and pericytes with contractile potential encircling descending vasa recta. These results suggest that included in the complex hemodynamic adjustment to chronic sodium depletion is a significant reduction in inner medullary blood flow that may be important in maintaining enhanced papillary solute concentration. In addition, the anatomy of the medullary vasculature is compatible with regional regulation of medullary blood flow.
Nephron heterogeneity of phosphate reabsorption: effect of parathyroid hormone
Authors: Haramati A. , Haas J. A. , Knox F. G.
Published: December 2017

We evaluated the response of superficial and deep nephron proximal tubules to PTH in thyroparathyroidectomized (TPTX) rats fed a normal phosphate diet (0.7%). As phosphate reabsorption is not detectable in the ascending limb of the loop of Henle, fractional phosphate delivery (FDPi%) to the superficial early distal tubule and papillary loop of Henle reflects delivery from superficial and deep nephron proximal tubules, respectively. Re-collection micropuncture experiments were performed in nine acutely TPTX rats before and after the infusion of PTH (33 U/kg bolus; 1 U X kg-1 X min-1). In response to PTH, fractional phosphate excretion increased from 3.3 to 26.2% (P less than 0.05). FDPi% was less from the deep than from the superficial proximal tubule (5.7 vs. 15.7%, P less than 0.05) prior to PTH, indicating enhanced phosphate reabsorption by deep compared with superficial proximal tubules. During PTH infusion, FDPi% was increased in both nephron groups compared with control (P less than 0.05), but there were no differences in phosphate delivery between deep (28.0%) and superficial (29.7%) proximal tubules. We conclude that in acutely volume-expanded TPTX rats, infusion of a pharmacologic dose of PTH decreases phosphate reabsorption in both superficial and deep nephrons. Furthermore, the heterogeneity of FDPi% from deep compared with superficial proximal tubules seen in TPTX rats is absent during PTH infusion.
Sympathetic nervous system in the loss of autoregulation in acute renal failure
Authors: Kelleher S. P. , Robinette J. B. , Conger J. D.
Published: December 2017

The responsiveness of the renal vascular system was investigated in uninephrectomized Sprague-Dawley rats in which acute renal failure had been induced by norepinephrine. The animals were studied at 1' and 3 wk after norepinephrine infusion. Uninephrectomized littermates served as controls. Compared with controls, there was an absence of renal blood flow autoregulation in 1-wk acute renal failure that returned in part by 3 wk. In 1-wk rats there was a marked increase, rather than decrease, in renovascular resistance as renal perfusion pressure was decreased. The renal vasculature was significantly less responsive in 1-wk rats than in control or 3-wk animals when acetylcholine, angiotensin II, or norepinephrine was infused into the renal artery at minimal vasoactive doses (all P less than 0.01). Paradoxically, renal vasoconstriction in response to renal nerve stimulation was greater in 1-wk than in 3-wk and control rats (P less than 0.01) and was not inhibited by renal artery infusion of phenoxybenzamine. Renal denervation significantly improved renal blood flow autoregulation in 1-wk animals (P less than 0.001) and completely abolished the increase in renovascular resistance as renal perfusion pressure was lowered. No effects of renal denervation on renal blood flow autoregulation were seen in control and 3-wk rats. It is concluded that renovascular responses to neurohumoral stimuli are aberrant in acute renal failure. The loss of renal blood flow autoregulation is related to an increased renovascular resistance that is due to increased activity of non-alpha-adrenergic mechanisms of the autonomic nervous system.
Heterogeneity of uridine incorporation along the rabbit nephron. II. Effect of DOCA
Authors: Vandewalle A.
Published: December 2017

Uridine uptake was investigated in microdissected segments from the entire length of tubules from normal rabbit kidneys and the effect DOCA treatment was studied. Uridine uptake was measured after in vitro incubation of kidney pyramids with [14C]uridine (0.25 mM). Determinations were done on intact segments, and also using TCA precipitation on both the acid-soluble fraction considered as representative of the pyrimidine nucleoside precursor pool and the acid-insoluble one containing the precipitated nucleic acids. In normal rabbits the fraction of uridine incorporated in the insoluble fraction represented about 5% of that in the soluble one for all segments. Maximal values were found in the distal convoluted tubule (DCT) and the connecting tubule (CNT), where uridine uptake was two- to fourfold higher than in other segments. After DOCA treatment, a significant increase in uridine uptake in the soluble fraction was present in the cortical collecting (CCT) (+90%, P less than 0.001) and medullary collecting tubules (MCT) (+63%, P less than 0.05) when values were corrected for the cellular swelling induced by DOCA in all segments. An even more marked rise in uridine incorporation was observed in the insoluble fraction of the DCT (+88%, P less than 0.05), CCT (+291%, P less than 0.001), and MCT (+422%, P less than 0.05). These results indicate that 1) the level of uridine uptake in soluble and insoluble fractions widely differs along the tubular length of the normal rabbit nephron; and 2) DOCA treatment induces an increase of uridine incorporation in both the pyrimidine precursor pool and nucleic acids of the collecting tubule, known to be the target site for corticosteroid hormones.
Tetraethylammonium transport by isolated perfused snake renal tubules
Authors: Hawk C. T. , Dantzler W. H.
Published: December 2017

Tetraethylammonium (TEA) transport was studied in isolated perfused snake (Thamnophis spp.) proximal renal tubules. Unidirectional lumen-to-bath (J1----bTEA) and bath-to-lumen (J1----bTEA) fluxes exhibited saturation kinetics, but Jb----1TEA also exhibited an apparent diffusive component and J1----bTEA did not. Jb----1TEA exceeded J1----bTEA at all concentrations studied, resulting in net TEA secretion. Transport into cells across both luminal and peritubular membranes was apparently against an electro-chemical gradient and was inhibited by cyanide. Km for J1----bTEA (5.9 microM) was about one-third Km for Jb----1TEA (19.9 microM), indicating greater affinity of the luminal transporter for TEA; but Vmax for Jb----1TEA (153 fmol X min-1 X mm-1) was about six times Vmax for J1----bTEA (27 fmol X min-1 X mm-1), indicating a greater capacity of the peritubular transporter for TEA, which could account for net TEA secretion. Jb----1TEA was inhibited by N-methylnicotinamide (NMN) in the bath, but J1----bTEA was inhibited initially and then apparently transstimulated by NMN in the lumen, indicating possible countertransport. J1----bTEA, but not Jb----1TEA, was significantly reduced by replacement of sodium with sucrose, indicating possible sodium dependency of the luminal transporter. All data indicate active (either primary or secondary) TEA transport at both luminal and peritubular membranes but net transepithelial transport in the bath-to-lumen direction.
Aldosterone effects on papillary collecting duct pH profile of the rat
Authors: Higashihara E. , Carter N. W. , Pucacco L. , Kokko J. P.
Published: December 2017

A number of recent studies have suggested that the medullary and papillary collecting ducts possess a high capacity for bicarbonate reabsorption. Because 3H binding studies have suggested the presence of mineralocorticoid receptors along the papillary collecting duct and because clearance studies have suggested that aldosterone affects urinary acidification, we designed a series of clearance and micropuncture studies to examine the role of the papillary collecting duct in final urinary acidification. The studies were conducted in intact and adrenalectomized rats receiving either replacement glucocorticoids alone or glucocorticoids with acute infusion of physiological doses of aldosterone. The results show that under normal acid-base status all rats had a pH profile along the papillary collecting duct with more terminal portions being more acidic than the proximal portions. This decreasing pH profile was diminished in acidotic adrenalectomized rats but returned to normal with acute administration of aldosterone. In all cases the luminal potential difference was slightly positive and did not change with adrenalectomy or administration of aldosterone. These results demonstrate that the papillary collecting duct participates in the generation of maximal urinary pH gradient and that this process is partly influenced by aldosterone.
Binding of [3H]bradykinin in isolated nephron segments of the rabbit
Authors: Tomita K. , Pisano J. J.
Published: December 2017

Binding of intrinsically labeled [3H]bradykinin was studied in isolated nephron segments of the rabbit. Highest binding was observed in the cortical collecting tubule (5.76 +/- 0.34 X 10(-18) mol/mm) and the outer medullary collecting tubule (5.24 +/- 0.25 X 10(-18) mol/mm, means +/- SE, n = 6). Small but significant binding was also seen in the glomerulus, proximal straight tubule, cortical thick ascending limb of Henle's loop, and distal convoluted tubule. Lysyl-bradykinin, methionyl-lysyl-bradykinin, and tyrosine-8-bradykinin (but not des-arginine-9-bradykinin, vasopressin, angiotensins I and II, or prostaglandins) competed with [3H]bradykinin. The site of highest kinin binding (collecting tubule) is downstream from the highest concentration of kallikrein (granular portion of distal convoluted tubule). The binding data indicate that the major sites of kinin action in the kidney are the cortical and medullary collecting tubules. One action of kinins could be the stimulation of prostaglandin synthesis in the collecting tubules, which are known to actively synthesize prostaglandins.
Transport and metabolism of glucose by renal proximal tubular cells in primary culture
Authors: Sakhrani L. M. , Badie-Dezfooly B. , Trizna W. , Mikhail N. , Lowe A. G. , Taub M. , Fine L. G.
Published: December 2017

A highly purified suspension of rabbit proximal tubules was cultured in a hormone-supplemented serum-free medium. This primary culture yielded a homogeneous population of cells that demonstrated functional and morphological polarity in mono-layers. The characteristics of the Na-dependent glucose transporter in the luminal membrane were studied by measuring the uptake of alpha-methylglucoside (AMG). The kinetics of Na-dependent AMG uptake were consistent with a single saturable system with an apparent Km of 0.8 mM and Jmax of 0.14 nmol X mg-1 X min-1. AMG permeability was 0.10 microliter X mg-1 X min-1. Uptake was inhibited 95% by 0.1 mM phlorizin and by removal of sodium. The stoichiometry of Na/glucose interaction with the carrier was 2:1. These characteristics are typical of the characteristics described for the late proximal tubule. To examine whether the glucose that enters the cell across the luminal membrane is incorporated into the metabolic pool of the cell, we studied the oxidation of [14C]glucose to 14CO2 in the absence and presence of phlorizin. Significant decarboxylation of [1-14C]glucose and [6-14C]glucose was observed, consistent with the existence of aerobic metabolism and a hexose monophosphate shunt. In the presence of 0.1 mM phlorizin, uptake and oxidation of D-glucose were inhibited to an identical degree, suggesting that luminal uptake is a rate-limiting step in the oxidation of glucose by these proximal tubular cells. These studies indicate that proximal tubular cells in primary culture utilize glucose as an energy source and that the glucose derived from transport across the luminal membrane is incorporated into the metabolic pool of the cell.