bluuebunny/crossref_metadata_2025 huggingface dataset:
Bulk flow of cerebrospinal fluid into brain in response to acute hyperosmolality
Authors: Pullen R. G. , DePasquale M. , Cserr H. F.
Published: December 2017

Brain volume is regulated during acute hyperosmolality based, in part, on the tissue gain of Na, Cl, and K. This study evaluates the contribution of bulk flow of cerebrospinal fluid (CSF) into brain to the volume regulatory gain of electrolytes. Artificial CSF containing radiolabeled albumin and diethylenetriamine penta-acetic acid (DTPA) was perfused for 60 min through the ventricles and/or subarachnoid space of anesthetized rats, and tracer clearances from CSF to brain were measured as a function of plasma osmolality. Osmolality was elevated after 30 min of perfusion by intraperitoneal injection of hypertonic NaCl or mannitol. Albumin and DTPA clearances increased with osmolality at the same rate, despite a sevenfold difference in diffusion coefficient, consistent with osmotic stimulation of a bulk flow component of tracer influx into brain. The volume shift estimated on the basis of this data is 114 microliters CSF/g dry wt brain for a 60-mosmol/kg increase in osmolality. Results indicate that CSF is a major source of the volume regulatory gain of Na and Cl, but not of K.
Postnatal maturation of rabbit renal collecting duct: intercalated cell function
Authors: Satlin L. M. , Schwartz G. J.
Published: December 2017

Intercalated cells play a major role in renal regulation of acid-base balance. We used fluorescent dyes to characterize postnatal maturation of intercalated cells. We stained rabbit collecting ducts with the pH-sensitive dye 6-carboxyfluorescein diacetate and identified individual intercalated cells by their bright green fluorescence. Number of fluorescent cells per millimeter tubule doubled during maturation in midcortex (68 +/- 7 to 121 +/- 9; P less than 0.01) but did not change in outer stripe of outer medulla. Excitation-ratio (490/450 nm) fluorometry of individual cells in nonperfused tubules revealed an increase in pH of cortical intercalated cell from 7.28 +/- 0.03 in newborn to 7.43 +/- 0.03 in adult (P less than 0.005); principal cell pH did not change with age, averaging 7.10 +/- 0.05. The smaller difference in pH between intercalated and principal cells in neonates suggested a paucity of H+ pumps in immature intercalated cells. Indeed, few cortical intercalated cells trapped the weak base acridine orange in cytoplasmic vesicles that contained H+ pumps or demonstrated selective uptake of 3,3'+-dipentyloxacarbocyanine, a fluorescent cation that probes for mitochondrial potential. Intercalated cells in neonatal medullary collecting ducts had a cell pH similar to that measured in the adult, as well as numerous acidic cytoplasmic vesicles and significant mitochondrial potentials. In conclusion, intercalated cells in cortical collecting duct underwent postnatal proliferation and maturation, whereas those cells present in outer medullary collecting duct, where proliferation was virtually complete by 1 wk of age, were nearly differentiated. Signals directing this centrifugal pattern of postnatal renal maturation are presently unknown.
Possible modulatory role of angiotensin II on atrial peptide-induced natriuresis
Authors: Salazar F. J. , Granger J. P. , Fiksen-Olsen M. J. , Bentley M. D. , Romero J. C.
Published: December 2017

Studies showing that atrial natriuretic peptides (ANP) induce a suppression of the renin-angiotensin system suggest that there might be a modulatory influence of angiotensin II (ANG II) on the natriuretic effect of the ANP system. To evaluate this possibility we assessed, in anesthetized dogs, the net increments in fractional excretion of sodium (FENa) and lithium (FELi) produced by ANP and by the inhibition of ANG II formation with captopril. These agents were infused at separate time periods into the renal artery at a maximal level that has been shown not to alter glomerular filtration rate (GFR). ANP caused an increment in FENa of 4.0 +/- 0.2, whereas captopril caused a much smaller increase of 0.2 +/- 0.04, indicating that most of the natriuretic effect of ANP is unlikely to be solely accounted for by inhibition of ANG II. The administration of both ANP and captopril produced increases in the FELi used as a marker for proximal tubular reabsorption. An infusion of ANG II superimposed on the infusion of captopril reduced the FENa from 1.5 +/- 0.3 to 0.8 +/- 0.1. Under these conditions the administration of ANP produced an increment of 2.7 +/- 0.4 in the FENa. This increase in FENa is 32.5% less than the net increase obtained when ANP was given in the absence of ANG II, whereas under these conditions FELi remained statistically unchanged. These results suggest that the modulatory activity of ANG II on the natriuretic affect of ANP could be negligible under normal conditions.
Effect of acute renal failure on insulin disposition in the isolated perfused rat kidney
Authors: Brier M. E. , Aronoff G. R. , Mayer P. R.
Published: December 2017

The renal disposition of insulin in acute renal failure has not been evaluated. We used the isolated perfused rat kidney to test the hypothesis that acute renal failure (ARF) decreases renal insulin clearance. We used warm ischemia for 45 min, uranyl nitrate 5 mg/kg, ureter ligation, and nonfiltering kidneys as methods of inducing ARF. Comparisons were made with normal control kidneys. The concentrations of insulin in perfusate and urine was determined by radioimmunoassay. Acute renal failure caused significant reductions in glomerular filtration rate, sodium and potassium reabsorption, and an increased urine pH. Warm ischemia and uranyl nitrate toxicity caused a 50% decrease in the renal clearance of insulin. Nonfiltering kidneys cleared insulin at a rate 90% decreased from controls. Ureteral ligation caused a 32% decrease in insulin clearance. Filtration was necessary for insulin to be cleared from perfusate. We conclude that ARF decreased renal insulin clearance through a decrease in insulin uptake from both the tubular lumen and peritubular surface.
Detection of a Na+-H+ antiporter in cultured rat renal papillary collecting duct cells
Authors: Wall S. M. , Muallem S. , Kraut J. A.
Published: December 2017

To examine whether Na+-dependent H+ transport is present in the papillary collecting duct, changes in intracellular pH (pHi) were evaluated in cultured papillary collecting duct cells acidified to a pHi of 6.3 and then placed into Na+-free or Na+-containing solutions. pHi was determined from changes in the fluorescent signal of the pH-sensitive dye BCECF. pHi did not change significantly when cells were placed in tetramethylammonium chloride- or KCl-containing solutions; however, a significant rise in pHi occurred when acid-loaded cells were placed in solutions containing 140 mM NaCl. The Na+-dependent rise in pHi was blocked by high concentrations of amiloride, but was not affected by alterations in membrane potential across the cell. The rate of rise in pHi was a function of extracellular sodium concentration with a Km for Na+ of 30 +/- 12 mM (n = 6). The properties of this Na+-dependent H+ efflux supports the presence of a Na+-H+ antiporter in the papillary collecting duct.
Kinetics of bicarbonate transport in the early proximal convoluted tubule
Authors: Liu F. Y. , Cogan M. G.
Published: December 2017

Bicarbonate permeability and bicarbonate transport kinetics in the S1 segment of the proximal convoluted tubule (PCT) have not been previously studied. In vivo microperfusion at a rate of 30 nl/min was performed in early and late PCT of Munich-Wistar rats. Bicarbonate permeability was first defined, using a bicarbonate-free, acetazolamide-containing perfusate, and was over fourfold higher in the early compared with the late PCT (20.4 +/- 1.8 vs. 4.6 +/- 0.4 X 10(-7) cm2/s, P less than 0.001). Net bicarbonate absorption was then measured using perfusate bicarbonate concentrations of 15, 25, 40, and 100 mM at 30 nl/min perfusion rate. Proton secretory rate was calculated for each group by subtracting the passive bicarbonate transport component from the net flux. Saturation kinetics of acidification were observed in both the early and late PCT. The maximal proton secretory rate at the highest luminal bicarbonate concentration (Vmax) in the early PCT was about twice that in the late PCT (504 +/- 37 vs. 265 +/- 15 peq.mm-1.min-1, P less than 0.001). However, the luminal bicarbonate concentration eliciting half-maximal proton secretion (apparent Km) was approximately the same (11 mM) in the early and late PCT. In conclusion, the early PCT has a higher bicarbonate permeability and proton secretory capacity than the late PCT. Increased Vmax but axial constancy of Km suggests that there is amplification of similar transport mechanism(s) affecting bicarbonate absorption in S1 compared with S2 cells.
Characterization of alpha-adrenoceptors on isolated rabbit renal arterioles
Authors: Edwards R. M. , Trizna W.
Published: December 2017

Postsynaptic alpha-adrenoceptors were characterized in afferent and efferent arterioles isolated from rabbit renal cortex. In both the afferent and efferent arteriole the selective alpha 1-adrenoceptor agonists phenylephrine and cirazoline produced concentration-dependent vasoconstrictor responses with the maximum responses being equal to that of norepinephrine. The selective alpha 2-receptor agonists B-HT 933 and UK 14304 were examined over a concentration range of 10(-9) to 10(-4) M. At these concentrations, they were either inactive (B-HT 933) or produced maximum contractile responses only 60% (UK 14304) of that produced by norepinephrine or the alpha 1-receptor agonists. Prazosin (10(-7) M), an alpha 1-receptor antagonist, produced a rightward shift in the concentration-response curve to norepinephrine, yielding apparent dissociation constants of 1.7 and 1.2 X 10(-8) M for the afferent and efferent arteriole, respectively. Prazosin also antagonized the contractile effects of UK 14304 (apparent dissociation constants, 0.9 and 1.1 X 10(-8) M for afferent and efferent arterioles, respectively). The selective alpha 2-receptor antagonist, rauwolscine (10(-7) M), had no effect on norepinephrine-mediated vasoconstriction. These results confirm the presence of alpha-adrenoceptors on the glomerular arterioles that mediated vasoconstriction. Furthermore, these receptors appear to be exclusively of the alpha 1-subtype.
Renal adrenergic effector mechanisms: glomerular sites for prostaglandin interaction
Authors: Pelayo J. C.
Published: December 2017

Micropuncture experiments were performed in Munich-Wistar rats to ascertain the renal microcirculatory sites at which prostaglandins interact with the renal nerve and angiotensin II. Renal nerve stimulation (RNS) of 3 Hz alone decreased single-nephron glomerular filtration rate (SNGFR) by 30%, the consequence of 10 and 35% reductions in the glomerular capillary hydrostatic pressure difference (delta P) and the single-nephron plasma flow (SNPF), respectively. Pre- and postglomerular vascular resistances increased. RNS during prostaglandin inhibition (indomethacin) resulted in a 70% reduction in SNGFR, secondary to 1) a further diminution in delta P and in SNPF, via heightened pre- and postglomerular vasoconstriction and 2) a marked decline in the glomerular ultrafiltration coefficient (LpA), from 0.058 +/- 0.006 to 0.027 +/- 0.002 nl.s-1.mmHg-1.g kidney wt-1 (P less than 0.005). Acute angiotensin II inhibition (MK-421 and [Sar1,Ala8]angiotensin II) in rats pretreated with indomethacin partially attenuated the effects of RNS on vascular resistances and therefore on delta P, SNPF, and SNGFR and prevented the reduction in LpA. Thus vasodilatory prostaglandins act as local modulators of both renal nerve and angiotensin II constrictive actions on glomeruli and renal microcirculation.
Mechanisms of H+ secretion by inner medullary collecting duct cells
Authors: Selvaggio A. M. , Schwartz J. H. , Bengele H. H. , Gordon F. D. , Alexander E. A.
Published: December 2017

Inner medullary collecting duct cells were isolated from rat papillae and grown to confluence on cover slips. H+ secretion was estimated by intracellular pH (pHi) changes measured with the fluorescent probe 2,7-biscarboxyethyl-5(6)-carboxyfluorescein. In buffered NaCl, pHi was 7.14 +/- 0.04 (n = 78). After acidification about 40% of monolayers exhibited Na+-independent alkalinization. In 5 mM glucose, cell alkalinization occurred at a rate of 47 +/- 4 nM H+/min. However, cell alkalinization did not occur in the presence of 2-deoxy-D-glucose (5–15 mM), iodoacetate (5 mM), or KCN (5 mM). All monolayers tested exhibited amiloride-inhibitable Na+-dependent cell alkalinization that appeared to be a first-order kinetic process; Km [Na+] was approximately 52 mM and Vmax was approximately 250 nM [H+]/min. At a constant extracellular [Na+] (110 mM), Na+-dependent H+ efflux was a first-order function of pHi; Km for intracellular [H+] = 321 nM and Vmax = 182 nM H+/min. The data are consistent with the presence of a primary active H+ pump and a secondary active Na+ exchanger. The metabolic energy for the active H+ pump can be provided by glycolysis and oxidative phosphorylation.
Role of prostaglandins in protein-induced glomerular hyperfiltration in normal humans
Authors: Vanrenterghem Y. F. , Verberckmoes R. K. , Roels L. M. , Michielsen P. J.
Published: December 2017

In six healthy male volunteers we studied the effect of indomethacin on the renal hemodynamic adaptations to an acute oral protein load. Control and protein loading studies without and with indomethacin were performed under sustained water diuresis. In the control studies without indomethacin, creatinine (CCr) and inulin clearance (CIn) remained stable and p-aminohippurate (PAH) clearance (CPAH) and kaliuresis slightly decreased while natriuresis progressively increased. Urinary prostaglandin E2 (PGE2) excretion remained stable. Indomethacin had no effect on these control values except on natriuresis and PGE2 excretion. After protein loading CCr, CIn, and CPAH progressively increased to reach a peak 2 h after protein intake. Natriuresis, kaliuresis, and urinary PGE2 excretion also increased significantly. After indomethacin the peak increase of CCr, CIn, and CPAH during the 2nd h was significantly blunted. Indomethacin had no effect on the increased sodium and potassium excretion. The increase of PGE2 excretion was significantly attenuated by indomethacin. The results presented suggest that renal prostaglandins play at least a partial role in the renal hemodynamic adaptations to protein loading.
Renin and angiotensinogen gene expression in maturing rat kidney
Authors: Gomez R. A. , Lynch K. R. , Chevalier R. L. , Wilfong N. , Everett A. , Carey R. M. , Peach M. J.
Published: December 2017

To determine whether angiotensinogen (Ao) and renin are synthesized by the immature kidney and to assess the changes in intrarenal renin distribution that occur with maturation, the kidneys from 24 newborn and 12 adult Wistar-Kyoto (WKY) and spontaneously hypertensive rats (SHR) were processed for renin immunocytochemistry using a highly specific anti-rat renin antibody. Kidney renin and Ao relative mRNA levels (mRNA/total RNA) were detected by Northern and dot blot techniques, using full-length rat renin and Ao cDNAs. Renal renin concentration (RRC) was measured by radioimmunoassay of angiotensin I (ANG I) and expressed as ng ANG I.h-1.mg protein-1 in the incubation media. RRC was higher in newborn than in adult SHR (979 +/- 164 vs. 206 +/- 47) and WKY (573 +/- 69 vs. 297 +/- 74) (P less than 0.05). In the newborn kidneys of both rat strains, renin was distributed throughout the entire length of the afferent arterioles and interlobular arteries, whereas in the adult kidneys renin was confined to the classical juxtaglomerular position. With maturation, there was a decrease in the proportion of immunoreactive juxtaglomerular apparatuses and arterial segments that contained renin. Kidney renin mRNA levels were 7.9-fold higher in the newborn than in the adult animal. Ao mRNA was detected in the newborn and adult kidneys of both rat strains. This study demonstrates conclusively that both renin and Ao genes are expressed in the newborn kidney, providing evidence for a local renin-angiotensin system that is subjected to developmental changes.
Nicotinamide as a rapid-acting inhibitor of renal brush-border phosphate transport
Authors: Wu K. I. , Bacon R. A. , Al-Mahrouq H. A. , Kempson S. A.
Published: December 2017

Administration of nicotinamide to rats produces specific dose-dependent inhibition of Na+-dependent phosphate transport across the renal brush-border membrane (BBM) and an increase in urinary excretion of phosphate. The intracellular mechanism of action of nicotinamide is not well established. As a step in this direction, the present studies determined whether nicotinamide was a rapid- or slow-acting regulator of the BBM phosphate transport system. Nicotinamide (0.5 g/kg) inhibited Na+-dependent BBM phosphate transport under conditions when de novo protein synthesis was inhibited by cycloheximide (1.0 mg/kg). Furthermore, the degree of inhibition was not different from that achieved by nicotinamide alone, suggesting that the action of nicotinamide does not require de novo protein synthesis. Studies on the time course of the onset of nicotinamide action revealed inhibition of BBM phosphate transport within 1 h after injection of nicotinamide, even in rats pretreated with cycloheximide. The rapid response to nicotinamide and its independence of de novo protein synthesis characterize nicotinamide as a rapid-acting regulator of the Na+-dependent phosphate transport system in renal BBM.
Antidiuretic hormone moves membranes
Authors: Handler J. S.
Published: December 2017

This review focuses on events at the apical plasma membrane of toad urinary bladder and mammalian collecting duct as their permeability to water changes in response to antidiuretic hormone (ADH) and to its withdrawal. The major marker of the permeability change is observed in freeze-fracture electron microscopy of the apical plasma membrane and consists of a dramatic increase in membrane particle aggregates and, in toad bladder but not in collecting duct, in fused vesicles (aggrephores) that contain particle aggregates in their limiting membranes. Withdrawal of ADH is accompanied by endocytosis at the apical membrane, reflecting retrieval of water-permeable, particle aggregate-containing membrane. Covalent labeling of the external surface of the apical membrane of toad bladder identifies specific proteins that are present in the apical membrane only during the response to ADH. Proteins of the same molecular weights are also present in the retrieved membrane when ADH is withdrawn. Several controversial areas are considered, including the extent of cell swelling as water flows across the epithelium from dilute apical solution to isotonic basal solution, whether only principal cells or principal cells and intercalated cells participate in the water permeability response of the collecting duct, the role of the cytoskeleton in the water permeability response, and the proposed second water permeability barrier that is affected by ADH, but not by adenosine 3',5'-cyclic monophosphate.
Low concentrations of ANP cause pressure-dependent natriuresis in the isolated kidney
Authors: Firth J. D. , Raine A. E. , Ledingham J. G.
Published: December 2017

The effect of alteration in renal perfusion pressure on the response of the isolated perfused rat kidney to concentrations of alpha-human atrial natriuretic peptide (ANP) within the pathophysiological range has been examined. At a perfusion pressure of 90 mmHg ANP concentrations of 50, 200, and 1,000 pmol/l were without effect on any parameter tested. At a perfusion pressure of 130 mmHg 50 pmol/l ANP produced an increase of 3.13 +/- 0.68 mumol/min in sodium excretion (UNa V), compared with a fall of 0.33 +/- 1.04 mumol/min in controls (P less than 0.02); fractional excretion of sodium (FENa) rose by 1.45 +/- 0.36% vs. -0.12 +/- 0.47% (P less than 0.05); glomerular filtration rate (GFR) was unchanged. At 200 and 1,000 pmol/l larger changes in UNa V and FENa were seen; only at 1,000 pmol/l was a significant effect on GFR observed. In contrast, frusemide (furosemide) at concentrations of 10 and 100 mumol/l was natriuretic at both 90 and 130 mmHg, with lesser absolute but greater proportional changes being seen at the lower pressure. It was concluded 1) the response of the isolated kidney to ANP is critically dependent on perfusion pressure, 2) at elevated levels of perfusion pressure the isolated kidney can respond to levels of ANP within the upper physiological and pathophysiological range.
Guanosine 3',5'-cyclic monophosphate as a mediator of inhibition of renin release
Authors: Henrich W. L. , McAllister E. A. , Smith P. B. , Campbell W. B.
Published: December 2017

The role of guanosine 3',5'-cyclic monophosphate (cGMP) as an inhibitory mediator of tissue renin release was examined in two different in vitro preparations. In rat superficial cortical slices, renin release stimulated by isoproterenol (10(-5) M) was ablated by atriopeptin III (ANP, 2.1 x 10(-8) M), nitroprusside (NP, 10(-3) M), and 8-bromoguanosine 3',5'-cyclic monophosphate (8-BrcGMP, 10(-3) and 10(-6) M). Arachidonic acid (10(-3) M)-stimulated renin release was also inhibited by ANP and 8-BrcGMP (10(-3) and 10(-6) M). Both ANP and NP increased tissue cGMP concentrations significantly (P less than 0.05), but neither had an effect on adenosine 3',5'-cyclic monophosphate (cAMP) concentrations. When methylene blue (10(-5) M), an inhibitor of guanylate cyclase, was added to slices incubated with isoproterenol and ANP, the inhibition of renin release by ANP was abolished. These results were confirmed in a preparation of isolated cultured rat juxtaglomerular cells. In these cells, isoproterenol induced a significant increase (58%, P less than 0.01) in renin release, which was inhibited by the addition of 8-BrcGMP (10(-6) M). These data demonstrate a direct inhibitory effect of ANP on isoproterenol- and arachidonic acid-induced renin release. The results with NP, 8-BrcGMP, and methylene blue suggest that cGMP is an intracellular mediator of this inhibition.
Renal effects of atrial natriuretic factor are independent of dopamine1 receptors
Authors: Murphy M. B. , Bass A. S. , Goldberg L. I.
Published: December 2017

This study examined whether the renal effects of atrial natriuretic factor (ANF) are mediated by dopamine1 (DA1) receptor activation. Intravenous infusion of low-dose ANF (0.0025 micrograms.kg-1.min-1) in euvolemic, pentobarbital sodium-anesthetized male mongrel dogs enhanced urine flow (V) by 71 +/- 14% (mean +/- SE) and urinary sodium excretion (UNaV) by 457 +/- 172% (P less than 0.05). Renal blood flow (RBF) was unchanged. Administration of pharmacological doses of ANF (0.1 microgram.kg-1.min-1) into the renal artery in volume-expanded dogs increased RBF by 26 +/- 6, V by 56 +/- 15, and UNaV by 101 +/- 42%. The selective DA1 receptor antagonist SCH-23390 (0.5 microgram.kg-1.min-1 iv) did not affect the response to ANF at either dose. The selective DA1 agonist, fenoldopam, increased RBF by 45 +/- 3, V by 94 +/- 27, and UNaV by 61 +/- 15% in volume-expanded dogs. With SCH-23390, fenoldopam increased RBF by only 16 +/- 6% whereas V and UNaV decreased by 16 +/- 10 and 17 +/- 10%, respectively. Accordingly, the failure of DA1 receptor-blocking doses of SCH-23390 to antagonize the response to ANF, at pharmacological or physiological doses, indicates that the renal effects of ANF, in the dog, are independent of DA1 receptor activation.
Model of renal cell volume regulation without active transport: role of a heteroporous membrane
Authors: Welling D. J. , Welling L. W.
Published: December 2017

Isolated proximal renal tubules of rabbit reach a passive steady-state volume in isotonic medium after active transport is inhibited by ouabain or by inhibition of cellular metabolism or lack of metabolic substrates. If the tubules are then placed in a hypotonic NaCl medium they swell rapidly and then exhibit a volume regulatory decrease (VRD) similar to that seen when active transport is present. We have mathematically modeled these transient events by assuming that the basolateral cell membrane is permeated by pores having at least two distinct reflection coefficients with respect to sodium, potassium, and chloride. VRD depends on the difference of the values of the reflection coefficients of the pore types. As hydrostatic pressure is exerted by the stretching basement membrane, water and ions can be expelled from the cells across the lower reflection coefficient pores and cause VRD. When the hydrostatic pressure compliance is removed, the cells fail to volume decrease unless sufficient extracellular impermeant solute is present to provide an osmotic force for water and ion exit. We conclude that a heteroporous membrane may be an essential feature for cell volume regulation and maintenance.
Glomerular uracil nucleotide synthesis
Authors: Cortes P. , Dumler F. , Levin N. W.
Published: December 2017

The biosynthesis of basement membrane material requires the sugar derivatives of uridine 5'-triphosphate (UTP) for protein glycosylation. Uridine and orotate utilization for the biosynthesis of uracil ribonucleotides was studied in isolated rat glomeruli incubated in vitro. At a 1 microM concentration total orotate utilization was 9.6 +/- 1.8 pmol.min-1.mg DNA-1 (1 mg DNA approximately 0.175 X 10(6) glomeruli), 51% of the total amount metabolized was used in ribonucleotide formation, and there was a significant UTP accretion. Except at a high initial concentration (50 microM), exogenous uridine failed to increase the UTP pool due to rapid uridine breakdown by a cytosolic phosphorylase. Inhibition of this enzyme with benzylacyclouridine resulted in increased biosynthesis and accretion of UTP, and in a 17-fold higher concentration of uridine, primarily produced from performed sources of nucleosides. Continuous addition of exogenous uridine to maintain its concentration at 1 microM resulted in a total uridine utilization of 550 +/- 30 pmol.min-1.mg DNA-1. Uridine salvage for ribonucleotide biosynthesis was only 3% of the total metabolized. In contrast to uridine, and presumably due to UTP pool compartmentation, orotate incorporation into uridine 5'-diphosphosugars was prominent. The metabolism of exogenous orotate was not decreased by the presence of large amounts of uridine and by an expanded UTP pool. It is concluded that when exogenous orotate is present, it is an important precursor for glomerular uracil ribonucleotide biosynthesis. Due to its rapid rate of catabolism, uridine cannot maintain ribonucleotide biosynthesis at a rate sufficient to result in UTP accretion unless it is provided continuously in substantial quantities.
Interactions between angiotensin II and renal nerves during chronic sodium deprivation
Authors: Mizelle H. L. , Hall J. E. , Woods L. L.
Published: December 2017

The aim of this study was to examine the importance of the renal nerves in mediating the sodium-retaining actions of angiotensin II (ANG II) during chronic sodium deprivation. In seven female dogs the left kidney was denervated and the urinary bladder was split, allowing continuous urine collection from separate innervated and denervated kidneys in the same dog. The dogs were maintained on a low-sodium diet (7 meq/day) for 9 days and then infused with the converting-enzyme inhibitor captopril (CAP, 14 micrograms.kg-1.min-1, 7 days) followed by CAP plus ANG II (10 ng.kg-1.min-1, 7 days). Mean arterial pressure (MAP) fell from a control of 88 +/- 4 (average for 5 days preceding CAP) to 65 +/- 3 mmHg during CAP. Infusion of ANG II along with CAP restored MAP to 97 +/- 5 mmHg. There were no significant differences in urine volume or sodium, potassium, chloride, or osmolar excretions between innervated and denervated kidneys during the control period, CAP infusion, or CAP plus ANG II infusion. Norepinephrine content was reduced by 99 +/- 1% in denervated kidneys. Because a differential response was not observed between innervated and denervated kidneys during ANG II blockade or infusion of ANG II, we conclude that the renal nerves do not play a major role in mediating the sodium-retaining effects of ANG II during chronic sodium restriction.
Phospholipase C activation by prostaglandins and thromboxane A2 in cultured mesangial cells
Authors: Mene P. , Dubyak G. R. , Abboud H. E. , Scarpa A. , Dunn M. J.
Published: December 2017

Phospholipase C activation by prostaglandins (PG) and thromboxane A2 (TxA2) was studied in cultured rat and human glomerular mesangial cells, measuring accumulation of radiolabeled inositol phosphates and cytosolic free calcium ([Ca2+]i) with the fluorescent intracellular probe fura-2. Prostaglandin F2 alpha (PGF2 alpha) and TxA2 were found to be the major eicosanoids active on this signaling pathway in rat and human cells, respectively, whereas other PG had lesser or no effects. PGF2 alpha and TxA2 rapidly induced accumulation of inositol trisphosphate accompanied by a simultaneous transient rise of [Ca2+]i, followed by sustained elevation or, in human cells, by a distinct second increase of [Ca2+]i within 45 s. A minor initial accumulation of inositol monophosphate was followed by marked elevation greater than 5 min after the early responses. Responses to different eicosanoids were mediated by separate receptors, functionally characterized using receptor antagonists or heterologous desensitization during sequential applications. Protein kinase C activation by serum and phorbol esters potently inhibited inositol phosphate accumulation and/or [Ca2+]i transients, indicating a pathway for a negative feedback on PG-evoked intracellular signals. We conclude that receptor-mediated phospholipase C activation underlies the biological effects of certain eicosanoids on the glomerular mesangium.
Diluting power of thick limbs of Henle. II. Bumetanide-sensitive 22Na+ influx in medullary vesicles
Authors: Reeves W. B. , Dudley M. A. , Mehta P. , Andreoli T. E.
Published: December 2017

We evaluated the effects of osmotic gradients on 22Na+ influx in vesicles prepared from rat outer renal medulla. 22Na+ influx driven in a coflow mode by an inwardly directed 100 mM KCl gradient was measured at 20 and 60 s; 1 mM bumetanide inhibited approximately 30% of 22Na+ influx. The bumetanide-sensitive 22Na+ influx was reduced by approximately 65% when either K+ or Cl- was omitted from the aqueous phases. We found that an osmotic gradient for vesicle shrinkage, that is, 600 mM urea in the extravesicular medium, enhanced the bumetanide-sensitive 22Na+ influx twofold. Conversely, an osmotic gradient for vesicle swelling, that is, with vesicles but not extravesicular media loaded with 600 mM urea, produced a 50% suppression of bumetanide-sensitive 22Na+ influx. Moreover, 600 mM extravesicular urea, an osmotic gradient for vesicle shrinkage, also reduced uptake of the nonspecific marker [14C]mannitol. These effects of osmotic gradients were not due to alterations in ionic driving forces, since bumetanide-sensitive 22Na+ influx driven in a counterflow mode by loading the vesicles with 100 mM NaCl also was activated or suppressed by osmotic gradients for vesicle shrinkage or swelling, respectively. We conclude that osmotic gradients, and/or vesicle volume changes, modulate bumetanide-sensitive Na+:K+:2Cl- activity.
Diluting power of thick limbs of Henle. III. Modulation of in vitro diluting power
Authors: Reeves W. B. , Molony D. A. , Andreoli T. E.
Published: December 2017

This paper considers the quantitative interplay of various factors in modulating diluting power of in vitro medullary and cortical thick ascending limbs of Henle (MTAL and CTAL, respectively) segments from mouse and rabbit. Experimentally, the measured diluting power of the in vitro rabbit CTAL is greater than that of the rabbit MTAL, although the inherent rate of net Cl- absorption at high perfusion rates is considerably greater in the rabbit MTAL than in the rabbit CTAL. Similar results apply when comparing the rabbit CTAL to the mouse MTAL exposed to antidiuretic hormone (ADH). Our calculations show that, in the rabbit CTAL, the measured static head luminal salt concentration can be accounted for quantitatively by the measured rate of net salt absorption at a high perfusion rate together with the passive permeability coefficients for Na+ and Cl-. Moreover, with perfusion rates of 10% of single-nephron glomerular filtration rate, the transport properties of the CTAL predict that, at the end of the CTAL, the static head luminal Cl- concentration occurs if the initial perfusate contains either 50 or 150 mM Cl-. Thus one can argue that, in vivo the CTAL may be the cardinal determinant of the TAL contribution to diluting power and to external salt balance. The relatively blunted diluting power of in vitro MTAL segments can be accounted for quantitatively by assuming that luminal dilution, and the attendant osmotic gradient from lumen to cell, suppresses the inherent rate of transcellular Cl- transport. Our calculations also show that prostaglandin E2 and peritubular osmolality increases blunt tubular diluting power. Thus in vivo, the MTAL segment may be the cardinal determinant of TAL contribution to concentrating power and to intrarenal salt balance.
Acidification adaptation along the inner medullary collecting duct
Authors: Bengele H. H. , McNamara E. R. , Schwartz J. H. , Alexander E. A.
Published: December 2017

Chronic acid feeding (ACD) stimulates and chronic alkali (AKL) feeding suppresses acid secretion along the inner medullary collecting duct (IMCD) of the rat. The purpose of these experiments was to determine whether these stimuli produce IMCD acidification adaptation. We tested this hypothesis by acutely changing systemic PCO2 in rats chronically fed ACD or ALK. Microcatheterization was used to measure pH and PCO2 and samples were simultaneously obtained for measurement of bicarbonate, titratable acid (TA), and ammonium. In 10 ACD rats (arterial pH, 7.26 +/- 0.01; PCO2, 88 +/- 1 mmHg) acid secretion along the IMCD was 506 +/- 88 nmol/min. In 10 ALK rats with similar arterial gases (pH, 7.16 +/- 0.02; PCO2, 82 +/- 1 mmHg) IMCD acid secretion was only 284 +/- 57 nmol/min, P less than 0.05. In ACD rats made hypocarbic (pH, 7.26 +/- 0.03; PCO2, 24 +/- 1 mmHg), IMCD acid secretion was 163 +/- 55 nmol/min. These data were compared with previously studied rats eating a regular diet. Acute hypocarbia (pH, 7.54 +/- 0.02; PCO2, 20 +/- 1 mmHg) completely suppressed acid secretion, 4 +/- 23 nmol/min, along the IMCD. We conclude that chronic alterations in acid-base status provide an IMCD "set" where comparable stimuli produce significant differences in IMCD acidification. These data provide additional support for the concept of IMCD acidification adaptation.
Effects of intrarenal adenosine on renal function and medullary blood flow in the rat
Authors: Miyamoto M. , Yagil Y. , Larson T. , Robertson C. , Jamison R. L.
Published: December 2017

Adenosine is a potent vasodilator of the systemic circulation. Infusion of adenosine into the aorta causes water and sodium retention and a fall in glomerular filtration rate and renal blood flow. The effect of adenosine on medullary blood flow is unknown. Because systemic vasodilatory effects may confound its renal actions, adenosine was infused into the renal artery of anesthetized Munich-Wistar rats at doses of 2, 6, and 15 micrograms/min. A marked dose-dependent increase in urinary flow and sodium excretion was observed. Inulin and p-aminohippuric acid clearance did not change significantly. Blood flow in vasa recta in the exposed renal papilla, as determined by fluorescence videomicroscopy, increased significantly only with the highest dose of adenosine. In control animals infused with the vehicle only, there was no change in any of the above variables. These results indicate that direct intrarenal infusion of adenosine in the rat increases urinary flow and sodium excretion and at higher doses also increases vasa recta blood flow. The effects on urinary flow and sodium excretion were therefore mediated by a mechanism other than an increase in vasa recta blood flow.
Effect of acute hyponatremia on rat brain pH and rat brain buffering
Authors: Adler S. , Simplaceanu V.
Published: December 2017

Acute hyponatremia was induced in rats by subcutaneous pitressin and intraperitoneal injection of 17.5 ml of 2.5% dextrose three times over 2 days. Brain pH was measured by use of 31P-nuclear magnetic resonance (31P-NMR). Two-hour infusion of a 500 meq sodium solution raised plasma sodium concentration (PNa) from 124.6 to 147 meq/l. Brain pH rose 0.1 pH units in the absence of changes in phosphocreatine or ATP. By contrast, increasing PNa in normal rats from 141 to 158 meq/l did not affect brain pH. Brain buffering was examined by 58-min exposure to 20% CO2 followed by a 38-min recovery. Intrinsic brain buffering (Bi) in hyponatremic and normal rats following 15 min of CO2 was similar, 49.1 and 45.5, respectively. After the next 40 min of CO2 exposure Bi was unchanged in hyponatremic rats at 45.8 but increased in controls to 102.5. In recovery Bi was higher initially in controls and rose significantly only in the controls. These results are consistent with the hypothesis that acute hyponatremia impairs the function of the Na+-H+ exchanger in brain.
Low-sodium environment induces adaptation in salamander diluting segments
Authors: Morgunov N. S. , Hirsch D. J. , Mobbs I. G.
Published: December 2017

The urodele Ambystoma tigrinum adapts to a distilled water environment by decreasing renal sodium excretion, but the site and mechanism of renal adaptation is unknown. Isolated diluting segments of Ambystoma kidney were studied after a 2-wk exposure of the animals to either distilled or artificial pond ([Na] = 1.2 meq/l) water. Identification of diluting segments was confirmed by electron microscopy. Morphometric study revealed evidence of increased tubular diameter and cellular hypertrophy in the distilled water group. Na+-K+-ATPase activity was increased in tubules from the distilled water group compared with tubules from the pond water group; 40.2 +/- 6.9 vs. 21.7 +/- 4.3 nM ADP generated.min-1.mm tubule length-1, P less than 0.036. This alteration in ATPase activity was due to an increase in the number of pump units present on the basolateral membrane, since specific ouabain binding was also doubled in distilled water compound with pond water group tubules, 20.7 +/- 2.3 vs. 9.1 +/- 0.9 fmol/mm tubule length tubules, P less than 0.011. An increase in transepithelial potential difference of the diluting segment was noted in distilled water group tubules (19.5 +/- 1.4 mV) compared with tubules in the pond water group (13.2 +/- 1.8 mV), P less than 0.015. We conclude that distilled water adaptation is associated with specific diluting segment structural and functional alterations, which are probably linked to an increase in sodium transport rate.
Phospholipids in signal transduction of mesangial cells
Authors: Mene P. , Simonson M. S. , Dunn M. J.
Published: December 2017

Glomerular mesangial cells respond to a variety of hormones, cytokines, and autacoids with an immediate hydrolysis of membrane phospholipids, including phosphatidylinositol, phosphatidylcholine, and phosphatidylethanolamine. Phospholipid metabolites such as inositol phosphates and diacylglycerol serve as initiating signals for subsequent intracellular events, including release of intracellular Ca2+ stores, increased conductance of the plasma membrane to various ions, changes of intracellular pH, and protein phosphorylation. The resting functional responses, including cell contraction and proliferation, are in turn regulated by simultaneous release of bioactive lipids such as eicosanoids and platelet-activating factor, which recognize receptors on both intrinsic glomerular cells and circulating, blood-borne cells. Through this complex signaling network, phospholipid metabolism plays a central regulatory role in mesangial functions in physiological conditions as well as in inflammatory or metabolic derangements of the kidney glomerulus.
Pathogenetic mechanisms in experimental hemoglobinuric acute renal failure
Authors: Zager R. A. , Gamelin L. M.
Published: December 2017

To evaluate mechanisms in hemoglobinuric acute renal failure (ARF) rats were infused with hemoglobin under aciduric or alkalinuric conditions. Aciduric rats developed azotemia, distal heme casts, and proximal tubular cell (PTC) necrosis, whereas alkalinuric rats developed no renal damage. Aciduria converted hemoglobin to met-hemoglobin, which precipitated, forming distal casts and inducing ARF. Hematin formation was not observed. The importance of met-hemoglobin production was indicated by its greater toxicity than hemoglobin during aciduria and by its ability to induce ARF even under alkalinuric conditions. A link between obstructing casts and PTC necrosis was identified; tubular obstruction induced by various mechanisms (met-hemoglobin casts; ureteral ligation; ischemic ARF) increased PTC hemoglobin uptake, producing lysosomal overload (giant endolysosomes) and PTC necrosis. This worsened ischemic ARF despite an otherwise subtoxic hemoglobin dose being used that had no discernible acute renal vasoconstrictive effect. Iron chelation (deferoxamine)/hydroxyl radical scavenger (Na benzoate) therapy did not mitigate this exacerbation of ischemic injury, suggesting a nonoxidant mechanism. We conclude that H is nephrotoxic, particularly when intratubular obstruction facilitates PTC heme uptake. Thus aciduria-induced met-hemoglobin cast formation and concomitant ischemic renal injury predispose to its nephrotoxic effect.
Modulation of natriuresis by sympathetic nerves and angiotensin II in conscious dogs
Authors: Persson P. B. , Ehmke H. , Kogler U. , Kirchheim H.
Published: December 2017

The effects of renal perfusion pressure and reflex sympathetic nerve stimulation on sodium excretion were studied in six conscious foxhounds on a normal sodium diet. This was done before, during common carotid occlusion (CCO), and during a recovery period following CCO. Three protocols were used 1) control (n = 6), 2) converting-enzyme inhibition (CEI, n = 6), and 3) CEI combined with a constant renal artery pressure (RAP, n = 5). In protocol 1, CCO increased RAP markedly (140.5 +/- 5.1 vs. 103.0 +/- 4.4 mmHg; P less than 0.001) along with a considerable natriuresis (128.4 +/- 20.1 vs. 86.3 +/- 15.1 mumol Na+/min; P less than 0.05). In protocol 2, CEI increased control sodium excretion but did not impair the natriuresis by CCO. Maintaining RAP at control levels in protocol 3 lead to an antinatriuresis (53.1 +/- 16.8 vs. 128.3 +/- 32.2 mumol Na+/min; P less than 0.05). Creatinine clearance was unaffected by all procedures. In conclusion, a change in ANG II formation shifts but does not impair the natriuretic response to CCO. A moderate sympathetic activation has a pronounced pressure-independent antinatriuretic effect, which is not mediated by angiotensin II.
Aldose reductase activities in microdissected rat renal tubule segments
Authors: Sands J. M. , Terada Y. , Bernard L. M. , Knepper M. A.
Published: December 2017

Osmoregulation in inner medullary cells depends in part on cellular accumulation of sorbitol, the production of which from glucose is catalyzed by aldose reductase. To identify nephron segments that contain aldose reductase, we developed a fluorometric ultramicroassay to measure aldose reductase activity in microdissected nephron segments from collagenase-treated kidneys of Sprague-Dawley rats. DL-Glyceraldehyde (10 mM) was used as a substrate. Substantial aldose reductase activities were found in all three inner medullary renal tubule segments: thin descending limbs, thin ascending limbs, and inner medullary collecting ducts. Activity increased with depth into the inner medulla in all three segments. When aldose reductase activities were normalized by cell volume the activities in the three inner medullary segments were similar. Little or no aldose reductase activity was measured in glomeruli or any cortical or outer medullary nephron segment. Both proximal convoluted and proximal straight tubules were found to have a substantial capacity to reduce DL-glyceraldehyde, but the finding of greater reductase activity with D-glucuronate (10 mM) than with D-xylose (10 mM) indicated that the activity was due to aldehyde reductase. Sorbitol dehydrogenase (measured by a similar ultramicroassay method) was present in substantial amounts in proximal tubules, but not in inner medullary collecting ducts. The overall pattern of enzyme activities is consistent with the proposed osmoregulatory role for sorbitol in all three inner medullary renal tubule segments.(ABSTRACT TRUNCATED AT 250 WORDS)
Clonal sublines that are morphologically and functionally distinct from parental OK cells
Authors: Cole J. A. , Forte L. R. , Krause W. J. , Thorne P. K.
Published: December 2017

Three clonal subpopulations of opossum kidney (OK) cells were derived from the parental line. The distribution of apical microvilli suggested that the OK cell line was heterogeneous. The clonal OK sublines appeared homogeneous as reflected by microvilli, which were uniformly distributed on the apical surface. Parathyroid hormone (PTH), forskolin (FSK), and prostaglandin E1 (PGE1) increased adenosine 3',5'-cyclic monophosphate (cAMP) formation in OK cells and all of the clones. PTH inhibited sodium-dependent phosphate transport in parental cells and in OK/B and OK/P clones with maximal effects appearing at 4, 2, and 1 h, respectively. PTH had no effect on phosphate transport in OK/H cells. FSK inhibited phosphate transport in parental cells and OK/B and OK/P clones but was relatively ineffective in OK/H cells. PGE1 decreased phosphate transport in OK/B and OK/P cells but was ineffective in the parental line and in OK/H cells. Phorbol 12-myristate 13-acetate, a potent inhibitor of phosphate transport in the parental OK cell line, had little effect in the clonal sublines. These clonal lines have remained phenotypically stable for 10 passages and should prove useful in studying the regulation of phosphate transport by PTH as well as addressing the question of whether PTH receptor subclasses exist which couple to cAMP and/or calcium effector systems in kidney cells.
Dietary protein intake modulates glomerular eicosanoid production in the rat
Authors: Don B. R. , Blake S. , Hutchison F. N. , Kaysen G. A. , Schambelan M.
Published: December 2017

The quantity of protein in the diet modulates glomerular function. To study the effect of dietary protein intake on glomerular eicosanoid production, rats were randomized to either a high- (40%) or low- (8.5%) protein isocaloric diet. Ten to fourteen days later glomeruli were isolated and incubated in the absence (basal) and presence (stimulated conditions) of arachidonic acid, and production rates of prostaglandin (PG) E2, PGF2 alpha, and thromboxane B2 (TxB2) were determined by direct radioimmunoassay. Under basal conditions, glomerular production of all three eicosanoids was significantly greater in rats ingesting the high-protein diet. Glomerular production of PGE2 and TxB2 was also greater in animals fed the high-protein diet in the presence of arachidonic acid, suggesting that glomerular cyclooxygenase activity was augmented. In contrast, ingestion of a high-protein diet was not associated with a significant increase in eicosanoid production by renal papillae or in TxB2 release by clotting blood. To investigate the potential role of the renin-angiotensin system in the dietary protein-induced modulation of glomerular eicosanoid production, rats ingesting a high- or low-protein diet were randomized to treatment with an angiotensin-converting enzyme inhibitor or no therapy. Enalapril attenuated the dietary protein-induced augmentation in glomerular eicosanoid production. This effect occurred only when administered in vivo, since the active metabolite enalapril did not alter PGE2 production by isolated glomeruli when added in vitro. Dietary protein intake also modulated glomerular eicosanoid production in three models of experimental renal disease in the rat (streptozotocin-induced diabetes mellitus, Heymann nephritis, and partial renal ablation).(ABSTRACT TRUNCATED AT 250 WORDS)