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F C Rector

Publications and source records attributed to F C Rector.

At least 37 records · Page 2Linked to original sources

Active and passive components of NaCl absorption in the proximal convoluted tubule of the rat kidney.

The active and passive components of NaCl absorption were examined in doubly perfused proximal convoluted tubules (PCT) of the rat kidney. When anion concentration gradients were generated by perfusing the lumen with a high chloride, low bicarbonate solution and the peritubular capillaries with a complete solution resembling plasma ultrafiltrate, volume absorption (JV) was 1.79 nl/mm/min and estimated chloride absorption (JCl) was 270 pEq/mm/min. When anion gradients were eliminated by perfusing the peritubular capillaries with a high chloride solution, JV was reduced to 0.91 nl/mm/min and JCl to 140 pEq/mm/min. These residual rates of absorption were reduced to zero by removing potassium from the perfusates. In the presence of anion gradients, removal of potassium reduced JV from 1.79 to 0.60 nl/mm/min and JCl from 270 to 90 pEq/mm/min. It is concluded that: (1) when PCT lumen are perfused with high chloride solution and the peritubular capillaries perfused with an ultrafiltrate-like solution, approximately 50% of NaCl and water absorption is passive, driven by the anion gradients, and 50% is active; (2) when PCT lumen and peritubular capillaries are both perfused with high chloride solution, anion gradients are absent and all NaCl absorption is active.

Animals↗

Increased Vmax for Na+/H+ antiporter activity in proximal tubule brush border vesicles from rabbits with metabolic acidosis.

Na+/H+ antiporter activity in renal brush border vesicles from control rabbits and rabbits made acidotic with 3 days of NH4Cl gavage was measured using the acridine orange method. Acidotic rabbits exhibited a significantly higher Vmax for antiporter activity (2.80 +/- 0.45 fluorescence units X s-1 X mg protein-1) compared with controls (1.31 +/- 0.13) but the Km for Na+ was unchanged (23.7 +/- 3.5 for acidotic, 19.1 +/- 3.2 mM for controls). When the Vmax for Na+/H+ antiporter activity was considered in relation to the degree of acidosis achieved in the experimental animals, there was a correlation (r = -0.75) between Vmax and plasma total CO2 concentration. Amiloride (100 microM) inhibited Na+/H+ exchange (Na+ = 90 mM) by 59 +/- 7% in both control and acidotic animals, indicating that the observed stimulation in Na+/H+ antiporter activity was not due to increased electrically coupled cation exchange. These findings suggest that the response of the proximal tubule to chronic metabolic acidosis involves an adaptive increase in the Vmax for Na+/H+ antiporter activity in the brush border membrane that is correlated to the degree of acidosis in the animals.

Acidosis↗

Axial heterogeneity in the rat proximal convoluted tubule. II. Osmolality and osmotic water permeability.

To assess whether proximal luminal fluid becomes hypotonic with respect to plasma, free-flow micropuncture measurements were made sequentially from the end-proximal tubule to Bowman's space in 10 tubules of hydropenic Munich-Wistar rats. Osmolality in Bowman's space was 2.8 +/- 0.3 mosmol less than in plasma. Tubular fluid osmolality fell along the tubule and by the end-proximal tubule was 7.5 +/- 0.7 mosmol/kg less than in plasma or 4.7 mosmol/kg less than in Bowman's space. Since luminal fluid became hypotonic, the reabsorbate was hypertonic. The transepithelial osmotic water permeability (Pf) was calculated using simultaneously measured water reabsorption rates. The osmotic gradient responsible for water reabsorption was assumed to be either lumen-to-reabsorbate or lumen-to-peritubular plasma, with a reflection coefficient for sodium chloride of 0.7-1.0. The Pf was then estimated to be between 0.2 and 2.0 cm/s in the first millimeter of tubule and to have fallen to 0.1-0.2 cm/s by the end of the tubule. In conclusion, luminal hypotonicity develops in the rat proximal convoluted tubule and must be considered as part of the osmotic driving force for water reabsorption.

Animals↗

Proton transport and cell function.

The past five years have witnessed an explosion of information on the many and varied roles of H+ transport in cell function. H+ transport is involved in three broad areas of cell function: (a) maintenance and alteration of intracellular pH for initiation of specific cellular events, (b) generation of pH gradients in localized regions of the cell, including gradients involved in energy transduction, and (c) transepithelial ion transport. These processes each involve one or more of several H+ translocating mechanisms. The first section of this review will discuss these H+ translocating mechanisms and the second part will deal with the cellular functions controlled by H+ transport.

Anions↗

Gas production after reaction of sodium bicarbonate and hydrochloric acid.

Ingestion of sodium bicarbonate has been implicated as one of the proximate causes of spontaneous gastric rupture. However, the volume and rate of gas released from the reaction of ingested sodium bicarbonate and gastric acid has not been previously studied in detail. We, therefore, developed an in vitro method for measuring gas release after addition of sodium bicarbonate to a solution containing hydrochloric acid. From the results of our studies, we conclude that even though hydrochloric acid and sodium bicarbonate react instantaneously, the resulting gas production is slow, mainly because CO2 produced from the dehydration of carbonic acid dissolves in water and is only slowly released into the gas phase. The major exogenous factors that determine the rate of gas release are the volume of the solution, the quantity of reactants, the air volume over the reaction mixture, the partial pressure of CO2 of the acid solution before the addition of bicarbonate, and the stirring rate. The presence of food, alcohol, and carbonic anhydrase had relatively little if any effect. Based on our results, we believe that ingestion of the recommended dose of sodium bicarbonate (one-half teaspoon) would result in only small amounts of sudden gas release, probably not enough to be an important factor in causing spontaneous gastric rupture. On the other hand, we measured the amount of sodium bicarbonate that people actually select to take for indigestion, and all exceeded the recommended dose. Some people selected doses of bicarbonate that would result in several hundred milliliters of gas release within 3 min; it seems likely that such injudicious ingestion of sodium bicarbonate, if taken when the stomach was distended with air, food, and liquid, could be an important factor in spontaneous gastric rupture.

Bicarbonates↗

Effect of potassium concentration on bicarbonate reabsorption in the rabbit proximal convoluted tubule.

The direct effects of acute changes in K+ concentration on HCO-3 (JnettCO2) and volume reabsorption (Jv) were examined in isolated perfused rabbit proximal convoluted tubules (PCT). Increasing ambient K+ concentration from 5 to 8 mM did not change JnettCO2 (94.5 +/- 16.1 vs. 98.8 +/- 17.7 pmol X mm-1 X min-1) or Jv (1.27 +/- 0.15 vs. 1.24 +/- 0.16 nl X mm-1 X min-1). In contrast, reducing ambient K+ concentration from 5 to 2 mM inhibited JnettCO2 by 22% and Jv by 29%. Reducing luminal K+ concentration from 5 to 0 mM with constant bath K+ concentration at 5 mM did not affect JnettCO2 or Jv. Further reductions in bath K+ concentration to 0.5 and 0 mM showed a similar dependence of both fluxes on K+ concentration. Half-maximum inhibition of JnettCO2 was obtained at 1.1 mM ambient K+ concentration and of Jv at 0.85 mM. At zero bath K+ concentration JnettCO2 was 6.6 +/- 2.5 pmol X mm-1 X min-1 and Jv was 0.03 +/- 0.04 nl X mm-1 X min-1. To determine whether this rate of acidification was significantly different from zero, we examined the ability of the PCT to generate tCO2 concentration gradients with zero bath K+ concentration at slow perfusion rates. The tCO2 concentration gradient generated (0.94 mM) was not different from that found when the perfusate was inserted directly into the collection pipette in the absence of a tubule (0.71 mM). These data are consistent with the view that HCO-3 reabsorption is totally dependent on the Na+-K+-ATPase pump system.

Animals↗

Sodium, bicarbonate, and chloride absorption by the proximal tubule.

Proximal tubules are lined with epithelial cells that contain Na-K-ATPase in their basolateral cell membrane. The luminal cell membrane contains transport proteins that couple movement of many solutes to the active transport of sodium. The cells are connected by low-resistance junctional complexes that permit passive movement of solutes via a paracellular shunt pathway. Acidification is mediated by a Na/H antiporter localized specifically in the luminal membrane and a chloride-independent, voltage-dependent bicarbonate exit process in the basolateral membrane. The rate of acidification is controlled by the pH of the luminal and peritubular fluids. Reabsorption of NaCl from the high-chloride, low-bicarbonate fluid in the late proximal tubule is approximately 40% passive and 60% active. In proximal straight tubules the active component is entirely by simple rheogenic sodium transport, with chloride absorption driven through the paracellular shunt pathway by the lumen-negative PD. In convoluted tubules the active component is primarily neutral, with both sodium and chloride transported in approximately equivalent amounts through the cell. The mechanisms for neutral NaCl transport across the luminal membrane and for chloride exit across the basolateral membrane are unknown. A reduction in peritubular Starling forces (hydraulic and oncotic pressures) suppresses net proximal reabsorption by two mechanisms: 1) increased paracellular permeability with modest backleak of solutes (bicarbonate, glucose, amino acids) whose luminal concentration falls below their plasma level, and 2) specific inhibition of active neutral transcellular transport of NaCl by reduced peritubular protein concentration by some mechanism other than inhibition of Na-K-ATPase.

Absorption↗

Flow dependence of proximal tubular bicarbonate absorption.

Rat proximal convoluted tubules were microperfused in vivo to examine the effect of flow rate on bicarbonate absorption. When tubules were perfused with 25 mM bicarbonate, increases in perfusion rate from 15 to 33 to 49 nl/min caused bicarbonate absorption to increase from 105 +/- 4 to 176 +/- 8 to 209 +/- 7 pmol X mm-1 X min-1, respectively. Only 15% of this stimulation could be attributed to a flow-induced increase in the measured axial luminal bicarbonate concentration profile. In addition, effects of flow on passive bicarbonate diffusion or convection could not account for the observed stimulation. When tubules were perfused with 58 mM bicarbonate (a concentration previously shown to achieve maximal rates of proton secretion), increasing flow rate from 15 to 49 nl/min did not stimulate bicarbonate absorption. Thus, when examined as a function of mean luminal bicarbonate concentration, increases in flow increased the rate of proton secretion without affecting the maximal rate. The data are most consistent with flow-dependent stimulation of bicarbonate absorption, secondary to flow-dependent changes in luminal bicarbonate concentration, occurring by two mechanisms: 1) flow-dependent increases in the measured axial luminal bicarbonate concentration profile and 2) flow-dependent decreases in radial luminal bicarbonate concentration gradients.

Absorption↗

Effects of extracellular fluid volume and plasma bicarbonate concentration on proximal acidification in the rat.

The effects of systemic bicarbonate concentration and extracellular fluid volume status on proximal tubular bicarbonate absorption, independent of changes in luminal composition and flow rate, were examined with in vivo luminal microperfusion of rat superficial proximal convoluted tubules. Net bicarbonate absorption and bicarbonate permeability were measured using microcalorimetry. From these data, net bicarbonate absorption was divided into two parallel components: proton secretion and passive bicarbonate diffusion. The rate of net bicarbonate absorption was similar in hydropenic and volume-expanded rats when tubules were perfused with 24 mM bicarbonate, but was inhibited in volume-expanded rats when tubules were perfused with 5 mM bicarbonate. Volume expansion caused a 50% increase in bicarbonate permeability, which totally accounted for the above inhibition. The rate of proton secretion was unaffected by volume expansion in both studies. The rate of net bicarbonate absorption was markedly inhibited in alkalotic expansion as compared with isohydric expansion. Bicarbonate permeabilities were not different in these two conditions, and the calculated rates of proton secretion were decreased by greater than 50% in alkalosis. Net bicarbonate absorption was stimulated in acidotic rats compared to hydropenic rats. This stimulation was attributable to a 25% increase in the rate of proton secretion. We conclude that (a) proton secretion is stimulated in acidosis, inhibited in alkalosis, and is not altered by volume status; (b) bicarbonate permeability is increased by volume expansion but is not altered by increases in plasma bicarbonate concentration; (c) when luminal bicarbonate concentrations are similar to those of plasma, net bicarbonate absorption is dominated by proton secretion and is thus sensitive to peritubular bicarbonate concentrations, and insensitive to extracellular fluid volume; (d) when luminal bicarbonate concentrations are low and proton secretion is slowed, bicarbonate permeability and thus extracellular fluid volume have a greater influence on net bicarbonate absorption.

Absorption↗

Evidence for coupled sodium/hydrogen exchange in the rat superficial proximal convoluted tubule.

Recent in vitro studies from the rat and rabbit have suggested a tightly coupled sodium/hydrogen ion exchanger on the luminal membrane of proximal tubules. The steep sodium gradient from the lumen to cell supplies indirect energy for hydrogen ions to be pumped from the cell to the lumen. However, a proton translocating pump has been demonstrated in other epithelia, which is independent of sodium transport and directly driven by ATP. To examine the role that sodium might play in the process of acidification, rat proximal convoluted tubules and their surrounding peritubular capillaries were perfused in vivo with artificial ultrafiltrate-like perfusion solutions. Total CO2 absorption was measured by microcalorimetry during alterations in sodium transport by replacement of the sodium with an impermeant cation, choline, or by inhibition of the (Na+ + K+)-ATPase by removing potassium from both perfusion solutions. Under control conditions the absolute rate of total CO2 absorption was 140 pmol/mm X min. In the choline substitution and potassium removal experiments, absolute total CO2 absorption fell to 23 and 28 pmol/mm X min, respectively. The data suggest that: 1) in the rat superficial proximal convoluted tubule approximately 80% of the bicarbonate absorption is tightly coupled to sodium transport; 2) this process is driven indirectly by the (Na+ + K+)-ATPase system; and 3) the residual 20% of acidification appears to be mediated by another mechanism or may be a consequence of technical limitations.

Absorption↗

Determinants of proximal bicarbonate, chloride, and water reabsorption during carbonic anhydrase inhibition.

To examine the magnitude and load dependency of proximal reabsorption during carbonic anhydrase inhibition with acetazolamide, Munich-Wistar rats were studied in hydropenia and following an increase in single nephron glomerular filtration rate (SNGFR) by either plasma or colloid-free Ringer expansion. During acetazolamide administration, when plasma loading increased SNGFR 50% compared with hydropenia, total CO2, chloride, and water proximal reabsorption rose proportionally, so that fractional proximal reabsorption rates remained constant (0.20-0.27). When SNGFR was comparably increased with Ringer expansion, total CO2, chloride, and water reabsorption were decreased relative to their respective rates during plasma expansion and, in fact, were not changed compared with hydropenic values. At all flow rates during carbonic anhydrase inhibition, end-proximal total CO2 and chloride concentrations were only slightly higher than in the glomerular ultrafiltrate, so that reabsorption was isohydric. In further studies, proximal reabsorption remained isohydric when the glomerular ultrafiltrate bicarbonate concentration was reduced in metabolic acidosis. In conclusion, absolute proximal reabsorption of bicarbonate, chloride, and water during carbonic anhydrase inhibition 1) proceeds isohydrically, with minimal anion concentration gradients generated, over a wide range of filtered loads, and 2) can be flow dependent (with plasma loading) as well as modulated by peritubular protein concentration, even though passive chloride transport is minimized.

Absorption↗

Proximal reabsorption during metabolic acidosis in the rat.

The mechanism by which proximal volume reabsorption is reduced during hyperchloremic metabolic acidosis was studied using free-flow micropuncture techniques in Munich-Wistar rats. Compared with control hydropenic conditions, absolute rates of proximal total CO2 and water reabsorption rates during NH4Cl-induced metabolic acidosis were diminished: from 557 +/- 35 to 204 +/- 19 pmol/min and from 13.0 +/- 1.0 to 9.7 +/- 0.6 nl/min, respectively. Inhibition of proximal volume reabsorption during metabolic acidosis was not attributable to alterations in the reabsorptive Starling forces, since peritubular capillary oncotic and hydraulic pressures were normal, or to acidemia itself, since acute respiratory acidosis was not found to decrease reabsorption. When partial repair of the acidosis was achieved by NaHCO3 infusion, absolute reabsorption of both total CO2 (390 +/- 48 pmol/min) and water (12.2 +/- 1.1 nl/min) significantly increased despite modest extracellular volume expansion. NaCl infusion in acidotic animals had no restorative effect on volume reabsorption. Mean values for single nephron glomerular filtration rate were similar under all conditions. Absolute chloride reabsorption tended to correlate better with absolute bicarbonate reabsorption and, hence, with the magnitude of the chloride concentration gradient developed than with the filtered chloride load. In conclusion, absolute proximal volume reabsorption during metabolic acidosis and its partial repair correlated with the absolute magnitude of bicarbonate filtered and reabsorbed. It is proposed that proximal volume reabsorption may be regulated, at least in part, by the anion composition of the glomerular ultrafiltrate.

Absorption↗

Effect of luminal bicarbonate concentration on proximal acidification in the rat.

The effect of luminal bicarbonate concentration on proximal tubular acidification was studied. Rat proximal convoluted tubules were perfused in vivo with solutions of varying bicarbonate concentration, and bicarbonate absorption was measured using microcalorimetry. Bicarbonate absorption was found to increase linearly with mean luminal bicarbonate concentrations up to 45 mM, but above this level it showed evidence of partial saturation. Bicarbonate permeability was measured and found to be 2.6 +/- 0.3 x 10(-7) cm2/s. Using this permeability, net bicarbonate absorption could be divided into two parallel components, both sensitive to luminal bicarbonate concentration: 1) proton secretion and 2) a passive bicarbonate leak. Proton secretion, when examined as a function of luminal bicarbonate concentration, exhibited saturation kinetics with an apparent Km of 16 mM and a Vmax of 200 pmol . mm-1 . min-1.

Animals↗

Effect of luminal and peritubular HCO3(-) concentrations and PCO2 on HCO3(-) reabsorption in rabbit proximal convoluted tubules perfused in vitro.

The effect of luminal and peritubular HCO3(-) concentrations and PCO2 on HCO3(-) reabsorption was examined in rabbit proximal convoluted tubules perfused in vitro. Increasing luminal HCO3(-) concentration from 25 to 40 mM without changing either peritubular HCO3(-) concentration or PCO2, stimulated HCO3(-) reabsorption by 41%. When luminal HCO3(-) concentration was constant at 40 mM and peritubular HCO3(-) concentration was increased from 25 to 40 mM without changing peritubular PCO2, a 45% reduction in HCO3(-) reabsorption was observed. This inhibitory effect of increasing peritubular HCO3(-) concentration was reversed when peritubular pH was normalized by increasing PCO2. Passive permeability for HCO3(-) was also measured and found to be 1.09 +/- 0.17 X 10(-7) cm2 s-1. Using this value, the passive flux of HCO3(-) could be calculated. Only a small portion (less than 23%) of the observed changes in net HCO3(-) reabsorption can be explained by the passive HCO3(-) flux. We conclude that luminal and peritubular HCO3(-) concentrations after HCO3(-) reabsorption by changing the active H+ secretion rate. Analysis of these data suggest that both luminal and peritubular pH are major determinants of HCO3(-) reabsorption.

Animals↗

Mineralocorticoid-resistant renal hyperkalemia without salt wasting (type II pseudohypoaldosteronism): role of increased renal chloride reabsorption.

A rare syndrome has been described in which mineralocorticoid-resistant hyperkalemia of renal origin occurs in the absence of glomerular insufficiency and renal sodium wasting and in which hyperchloremic acidosis, hypertension, and hyporeninemia coexist. The primary abnormality has been postulated to be a defect of the potassium secretory mechanism of the distal nephron. The present studies were carried out to investigate the mechanism of impaired renal potassium secretion in a patient with this syndrome. When dietary intake of sodium chloride was normal, renal clearance of potassium was subnormal (CK/GFR = 3.6 +/- 0.2%; normal subjects, 9.0 +/- 0.9%, N = 4) despite high normal or supernormal levels of plasma and urinary aldosterone. The fractional clearance of potassium remained subnormal (CK/GFR = 5.1 +/- 0.2%) during superimposed chronic administration of superphysiologic doses of mineralocorticoid hormone. Little increase in renal potassium clearance occurred when the delivery of sodium to distal nephron segments was increased further by the i.v. infusion of sodium chloride, despite experimentally sustained hypermineralocorticoidism. But potassium clearance increased greatly when delivery of sodium to the distal nephron was increased by infusion of nonchloride anions: sulfate (sodium sulfate infusion, low sodium chloride diet; CK/GFR = 63.7 +/- 0.4%) or bicarbonate (sodium bicarbonate plus acetazolamide infusion; CK/GFR = 81.7 +/- 1.7%). These findings indicate that mineralocorticoid-resistant renal hyperkalemia in this patient cannot be attributed to the absence of a renal potassium secretory capability or to diminished delivery of sodium to distal nephron segments; instead it may be dependent on chloride delivery to the distal nephron. We suggest that the primary abnormality in this syndrome increases the reabsorptive avidity of the distal nephron for chloride, which (1) limits the sodium and mineralocorticoid-dependent voltage driving force for potassium and hydrogen ion secretion, resulting in hyperkalemia and acidosis and (2) augments distal sodium chloride reabsorption resulting in hyperchloremia, volume expansion, hyporeninemia, and hypertension.

Adult↗