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A capacity for both net potassium absorption and net potassium secretion has been demonstrated in the inner medullary collecting duct. The quantitative importance, however, of the inner medullary collecting duct in the regulation of urinary potassium in potassium deficiency, however has not been established. To assess the contribution of this segment to potassium conservation, microcatherization studies were performed in male Sprague-Dawley rats maintained either on a control diet or on a potassium free diet for 72 h. In control animals approximately 15% of the filtered load of potassium was excreted. Analysis of tubule fluid along the inner medullary collecting duct failed to demonstrate evidence of net potassium movement. Administration of a potassium free diet resulted in a marked reduction in potassium excretion to 0.3% of the filtered load. In contrast to control the inner medullary collecting duct of experimental animals absorbed nearly 90% of the amount of potassium entering this segment, since fractional delivery to the terminal portion of the nephron was about 2%. These data indicate that the inner medullary collecting duct makes a significant contribution to maximal renal conservation of potassium, since previous studies have shown that only 5 to 10% of filtered potassium is present in the late distal tubule of surface nephrons in animals on a low potassium intake.

The purpose of this study was to characterize the alterations in collecting tubule Na+-K+-ATPase activity produced by sustained increments in dietary potassium in the rat and to evaluate the role of aldosterone in their generation. In adrenal-intact animals, feeding a high-potassium diet (10-fold that of control) or administration of a high physiological dose of aldosterone (5 micrograms X 100 g-1 X day-1), which simulates the delivery rate of this hormone during potassium loading (both for 7 days), caused marked increments in Na+-K+-ATPase activity in the cortical collecting tubule (CCT) but had no effect on the enzyme in the inner stripe of the medullary collecting tubule (MCT). A significant increase in enzyme activity was also observed after smaller dietary potassium increments (2.5 and 5 times the control) and after 4 (but not 2) days of dietary potassium load. In adrenalectomized rats provided with physiological replacement doses of corticosterone and aldosterone (0.8 micrograms X 100 g-1 X day-1), Na+-K+-ATPase activity in both CCT and MCT was similar to that of adrenal-intact controls but remained unchanged after 7 days on the potassium-enriched (10-fold) diet. In contrast, adrenalectomized animals receiving the high physiological dose of aldosterone displayed an increase in Na+-K+-ATPase activity of CCT comparable with that of adrenal-intact animals, whereas the enzyme activity in the MCT was unaffected. In conclusion, 1) following chronic potassium loading Na+-K+-ATPase activity increases significantly in the CCT with no change in its activity in the inner stripe of the MCT.(ABSTRACT TRUNCATED AT 250 WORDS)

To investigate the sites of action of aldosterone (Aldo) and chronic potassium loading (high K) and to determine whether dietary K can modulate collecting duct structure independent of Aldo, a morphometric study of the structure of the initial collecting tubule (ICT), the cortical collecting tubule (CCT), the medullary collecting duct in the outer stripe (MCDOS) and the inner stripe (MCDIS), and the papillary collecting duct (PCD) was carried out in a rat model in which plasma levels of Aldo were clamped at basal or physiologically elevated levels and high K could be induced without concomitant hyperaldosteronism. Elevated levels of Aldo increased cell area (Ac) and basolateral membrane surface (BLM) from control values in the ICT, CCT, and MCDOS but not in the MCDIS or PCD. High K with normal physiological levels of Aldo increased BLM in the ICT and CCT without a concomitant change in AC. No response was seen in MCDOS, MCDIS, or PCD. It is concluded that Aldo can modulate principal cell structure in the collecting duct system in segments extending from the ICT through the MCDOS in the rat and that high K independent of Aldo can influence collecting duct structure only in the ICT and CCT.

Chronic potassium loading results in an increased capacity of the distal nephron to secrete potassium. The cellular mechanism for this adaptation has been correlated to an increase in the activity of sodium-potassium-ATPase. Because adaptation may be dependent on the greater availability of potassium pumps in the basolateral membrane, a stereologic analysis of the membrane surface area was performed to determine whether the apparent increase in pump sites was due to an increase in cell membrane surface. With potssium adaptation, the number of microplicated cells in the outer medulla was reduced from 31 to 18%. There was a marked increase in the basolateral infolding of principal cells, and membrane surface increased by 32%. In papillary collecting duct cells, the basolateral membrane surface was unchanged but the surface density of the luminal membrane increased by 50%. These observations suggest that amplification of the basolateral cell membrane to increase the number of potassium pump sites per cell plays an important role in the mechanism of potassium adaptation in the outer medulla. A different mechanism involving the luminal membrane operates in the papillary collecting duct. Structural alterations in cell membrane surfaces are thus related to the regulation of the epithelial transport of electrolytes.

Studies were performed in the rat, under steady-state conditions, to determine the mode of potassium transport in the late distal tubule. In these experiments the potential difference and potassium activity were measured simultaneously in steady-state fluid droplets in control rats, during acute infusion of KCl, and in potassium-deficient rats. The results demonstrate active potassium secretion in the late distal tubule of the control and KCl-infused animals, since the observed potassium activity of 11.14 +/- 1.39 in control animals and 25.26 +/- 2.96 meq/liter in potassium-loaded animals was significantly greater than the predicted values of 7.75 +/- 0.59 and 14.01 +/- 0.94 meq/liter in the same groups, respectively. In potassium-deficient animals, however, the observed potassium activity in luminal fluid of 4.03 +/- 0.55 meq/liter was not different from the predicted value of 4.26 +/- 0.32. In summary, this study demonstrates that potassium secretion in the late distal tubule of the rat occurs by an active process and fails to demonstrate active absorption in the same tubular segment.

Studies were performed to 1) systematically compare Ling-Gerard microelectrodes and electrodes with large tips (3 to 5 mu O.D.) in the measurement of the transepithelial potential difference (PD) in the mammalian distal tubule and 2) to examine the sign and magnitude of the transepithelial PD along the length of the distal tubule. Recordings made with the tips of both types of electrodes placed together in the same tubule segment demonstrated no difference in transepithelial PD. Rapid influx of tubular fluid into the macroelectrode, however, was found to depolarize the tubular membrane, as recorded by both types of electrodes simultaneously. Technical critera were therefore established to prevent rapid influx of fluid and efflux of potassium chloride, filling the electrode, in the use of exploring electrodes with large tips. The sign of the transepithelial PD in early distal tubule was found to be negative and averaged -17.7 +/- 9.4 (mean +/- SEM) in Sprague-Dawley rats and approximately -12 mV in Wistar-Munich rats. The site of puncture was identified by dye transit time rate, microdissection, and cell morphology. The transepithelial PD increased along the length of the distal tubule and averaged -41.9 +/- 8.5 in the last portion accessible to examination.

Studies were performed in the rat to determine the transepithelial potential difference (PD) of the medullary collecting duct (MCD). With the microcatheter as an exploring electrode the PD from duct lumen to the reference electrode, placed in peritoneal fluid, was approximatley 1 mV, lumen positive, in the MCD. It was unlikely that the polarity and small magnitude of PD was due to an artifact created by the microcatheter, since essentially similar results were obtained by transmural puncture of the inner third of the duct using Ling-Gerard glass electrodes and glass microelectrodes, tip OD 3-5 micron. Moreover, a PD of a few millivolts, lumen positive, was observed in experimental states characterized by marked increases in potassium and hydrogen ion secretion and sodium absorption. Further studies showed that the diffusion potential existing within the interstitial fluid compartment, between papilla and reference electrode, was approximately +2 mV, owing to the higher NaCl concentration in the papillary area. These data demonstrate, therefore, that the transepithelial PD in MCD approaches zero in control animals, and is not substantially altered by marked changes in net electrolyte transport.

Studies were performed to examine some electrical properties of the mammalian distal tubule. Experiments were performed to evaluate the influence of changes in ion concentration in luminal fluid on transepithelial potential difference. Variations in the concentration of Na+ and K+, near the physiologic range, resulted in only modest changes in potential difference. During changes in Na+ the average slope, deltaV/ln C2/C1, was -1.52 +/- 0.39 mV and during variations in K+ the slope was -5.60 +/- 0.95 mV. Changes in Cl- concentration had no effect on transepithelial potential difference. Since the sum of transferase numbers did not equal 1, it seems likely that potential difference is influenced by a shunt current due to cellular rheogenic pumps. These data, therefore, indicate that variations in ion concentration in tubular fluid of the mammalian distal tubule do not significantly influence the resting transepithelial potential difference, and they suggest that analysis of permselective properties of the luminal cell membrane by transepithelial electrophysiologic approaches is probably invalid.

Previous studies have demonstrated that the rate of urinary potassium excretion is markedly greater in animals fed a high potassium diet than in animals on a control diet before and during the acute infusion of potassium salts. Because the contribution of the medullary collecting duct to urinary potassium under these conditions has not been determined, microcatheterization studies were performed to elucidate the role of that nephron segment in regulating potassium excretion in controls and in animals on a potassium-enriched diet. Although there was no evidence of net transport of potassium by the medullary collecting duct in control animals under basal conditions, net secretion of potassium by that nephron segment accounted for nearly one half of the increased rate of urinary excretion in animals fed a high potassium diet. During acute infusion of potassium salts, potassium secretion by the medullary portion of the collecting duct contributed approximately one half of the potassium in the final urine samples in both controls and animals fed a high potassium diet. Because the rate of urinary excretion was twofold greater in experimental animals during acute potassium loading, absolute potassium secretion in medullary collecting duct was twice the control rate in animals adapted to a high potassium dietary intake. These data demonstrate, therefore, that the medullary collecting duct plays an important role in regulating the excretion of potassium when the excretory load is either acutely or chronically increased.

Acute, reversible renal failure affecting primarily medullary structures, was produced by feeding rats a diet containing 5% oxonic acid and 2.5% uric acid for 7 days. Inulin clearance and selected tubular transport processes were studied and compared to controls. At the time of maximal renal injury, inulin clearance was reduced to 40% of control values, in association with a 60% reduction in urine concentrating ability, a 50% reduction in potassium excretion and a 56% reduction in ammonium excretion. During a 7-day recovery period, inulin clearance increased to 70% of control and potassium excretion returned to normal while the other tubular transport functions remained impaired. Thus, in acute renal failure, tubular functional impairments may recover at different rates.

Prevoius studies have suggested that the increase in specific activity of Na-K-ATPase in renal tissue during treatment with glucocorticoids occurred as a result of aconcurrent rise in net tubular reabsorption of sodium. Since recent data have indicated a specific effect of glucocortiocoids on epithelial cells, experiments were performed to determine whether enzyme activity and net sodium reabsorption could be dissociated. Evidence is provided demonstrating that base-line specific activity of Na-K-ATPase in rat renal cortex and outer medulla does not correlate directly with net sodium reabsorption since enzyme activity did not change after a chronic reduction in glomerularfiltration rate and the rate of sodium reabsorption. Further studies showed a markedrise in Na-K-ATPase after 4 days of treatment with methylprednisolone despite a fall in sodium absorption. These results suggest a direct effect of glucocorticoids onrenal Na-K-ATPase and illistrate the difficulty in assigning a transport role tothis enzyme from the correlation of specific activity with rates of net electrolyte transport.

Although clearance studies in man and experimental animals indicate that filtered lithium is reabsorbed primarily in the proximal tubule, it is unclear whether lithium is also reabsorbed in distal portions of the nephron. Micropuncture studies were, therefore, performed to determine the nephron sites involved in lithium transport during free flow. A method was established to estimate the concentration of lithium in nanoliter samples, using the Helium Glow photometer, which permitted the accurate measurement of lithium in tubular fluid samples over a range from 0.5--30.0 mM. Approximately 56% of filtered lithium and tubular fluid was reabsorbed at the end of the proximal convolution, while at the early distal tubule 75% of filtered lithium and water was reabsorbed. There was no change in net transepithelial movement of lithium beyond the loop of Henle. These data suggest that lithium transport is localized to the proximal tubule, including the pars recta. Lithium reabsorption does not occur in distal tubule or collecting duct. Beyond the early distal tubule net movement of lithium and sodium is dissociated.

Micropuncture studies were performed in rats infused with LiCl to induce stable plasma lithium concentrations of 2--3 mEq/l, or with an equivalent amount of NaCl. In free flow experiments LiCl reduced proximal tubule fractional reabsorption of sodium and potassium. Reduced reabsorption of bicarbonate, as reflected by a decrease in TF/PCl, was also observed. Proximal fractional reabsorption of chloride, however, was not affected. The TF/PIn at the end proximal tubule was 2.6 +/- 0.2 (mean +/- SEM) in controls and 2.1 +/- 0.1 in the experimental animals (P less than 0.025). In the distal portions of the nephron lithium treatment caused a fall in fractional reabsorption of water and sodium, while potassium secretion was stimulated in the distal tubule. Previous studies have indicated that lithium influences antidiuretic hormone stimulated water transport in the collecting duct. These experiments demonstrate that lithium also affects the transport of water and electrolytes in multiple nephron segments, including the proximal and distal convolution.

Studies were performed on the initial collecting tubule of the rat to determine whether potassium adaptation in this nephron segment is aldosterone-dependent. Previous studies demonstrated that chronic potassium loading, in animals with intact adrenal glands, caused an increase in transepithelial potential difference in late distal convolution, an increase in surface density of the basolateral cell membrane, SVBLM, of principal cells in the initial collecting duct, and a rise in plasma aldosterone levels. The present study shows that epithelial changes that characterize dietary potassium loading are not dependent on hyperaldosteronism, since potential difference (-47 +/- 1 vs. 40 +/- 3 mV, lumen negative) and SVBLM (2.91 +/- 0.11 vs. 2.53 +/- 0.09 micron2/micron3) increased significantly (P less than 0.05) in the late distal convolution of adrenalectomized, hormone-replaced animals in which plasma aldosterone levels were maintained at basal values of approximately 5 ng/dl. In addition, these experiments suggest that the initial collecting tubule is sensitive to the action of aldosterone, at physiological plasma levels, since chronic hyperaldosteronism, in the absence of potassium loading, increased SVBLM in initial collecting tubule cells. In contrast to other mineralocorticoid-sensitive tissues, however, neither the acute or chronic administration of aldosterone caused an increase in potential difference in late distal convolution. These results suggest that the mechanism by which aldosterone stimulates electrolyte movement is not identical in all target tissues.

The hypothesis that chronic metabolic acidosis encountered in some patients with primary hyperparathyroidism is due to inhibition of proximal HCO3 reabsorption has recently been challenged. Indeed, this action of parathyroid hormone (PTH) has only been observed in acute studies, whereas in animal models of chronic hyperparathyroidism a metabolic alkalosis has been induced, probably owing to the release of alkaline salts from bone tissue, and to the stimulation of tubular acid secretion by hypercalcaemia. Studies were, therefore, performed to determine the effect of PTH on the renal handling of HCO3 in an animal model in which changes in plasma calcium and phosphate, and nephrocalcinosis, all known to affect tubular acidification, did not occur. Thyroparathyroidectomized (TPTX) rats were infused with synthetic bovine hormone fragment bPTH 1-34 via Alzet minipumps at the rate of 0.7 U h-1 to simulate normal endogenous production of PTH (group I) and at the three-fold higher rate of 2.1 U h-1 (group II). In order to prevent changes in serum calcium and phosphate, and nephrocalcinosis, animals of group II were fed a Ca- and P-free diet prior to TPTX (compared with a regular diet for group I) and both groups were treated with dichloromethylene-diphosphonate (Cl2MDP, 10 mg kg-1 day-1) and a Ca-free diet during PTH infusion. During the course of PTH infusion both groups of animals had stable and normal levels of plasma calcium and phosphate.(ABSTRACT TRUNCATED AT 250 WORDS)

Acute infusions of isotonic saline in the rat cause an increase in glomerular filtration rate and in the excretion of salt and water. The kidney swells, due to expansion of tubular and interstitial volume. Despite the increase in tubular diameter, transit time through the proximal tubules and loops of Henle is decreased, presumably owing to a greatly accelerated rate of tubular flow. Proximal tubular reabsorption, measured in blocked tubules, is inhibited in a way that cannot be ascribed to changes in tubular diameter. The prolongation of proximal reabsorptive half-time is not affected by the administration of aldosterone. It occurs equally in rats chronically loaded with or deprived of salt, and it is therefore not likely that it is influenced by the renal content of renin. In contrast, reabsorption from the distal convoluted tubule is enhanced by saline infusion. This change is observed in segments of tubules blocked with oil and isolated from their glomeruli and thus appears to occur independently of changes in glomerular filtration or tubular flow.

The effect of increased hydrostatic pressure in the peritubular vessels on net sodium reabsorption from the proximal tubule was examined in the Necturus. An increase in the pressure gradient of 2.0 cm H(2)O across the wall of the proximal tubule, produced by ligation of the postcaval vein was associated with a marked reduction in net reabsorption and an increased back flux of water and electrolytes. This change was accompanied by a slight, but significant drop in the transepithelial electrical potential but not by an alteration in the steady-state chemical gradient. These studies highlight the importance of changes in the permeability characteristics of the proximal tubule on net sodium transport.

The renal response to chronic depletion of extracellular volume was examined using the techniques of micropuncture. Depletion of salt and water was produced by administration of furosemide to rats maintained on a sodium-free diet. There was a marked fall in body weight, plasma volume, and glomerular filtration rate. The intrinsic reabsorptive capacity of the proximal tubule, measured by the split-droplet technique, was greatly enhanced. The acceleration of proximal fluid reabsorption could not be accounted for by changes in filtration rate, tubular geometry, or aldosterone secretion. The half-time of droplet reabsorption in the distal tubule was not altered by sodium depletion. An increase in the reabsorption of fluid in the proximal tubule, as demonstrated directly in the present experiments, provides an explanation for a variety of clinical phenomena associated with volume depletion.