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Water and electrolyte transport in Henle's loop and distal tubule after renal sympathectomy in the rat.

The effect of acute renal denervation (AD) on transtubular movement of water and electrolytes in Henle's loop (LH) and distal tubule (DT) of superficial nephrons was studied by the technique of in vivo microperfusion in Inactin-anesthetized hydropenic male rats. In time-control experiments (n = 6) no changes in either whole kidney or single nephron function were detected. Predenervation results of denervation experiments (n = 10) were similar to those of the time-control series. AD resulted in increased urine, sodium, and potassium excretion in the absence of changes of either arterial blood pressure or GFR. Compared with predenervation data, AD led to significant decreases in net transtubular fluxes (J) of water and sodium in both perfused segments with delivery kept constant (delta JNa: LH = 144 +/- 26 peq X min-1, 9.1 +/- 0.3%; DT = 94 +/- 11 peq X min-1, 22.3 +/- 1.1%). Potassium absorption in the loop decreased but an increase in net addition (secretion) of K in the distal tubule on denervation occurred (delta JK = 8.7 +/- 1.9 peq X min-1, 68.7 +/- 3.2%). Use of the reperfusion technique gave similar results. These results indicate that efferent renal sympathetic nerve activity directly influences water, sodium, and potassium transport in the loop of Henle and distal tubule of the anesthetized rat.

Animals↗

Glucose transport in the short loop of Henle of the rat kidney. Its characterisation by transport constants.

Short loops of Henle in rat kidneys were perfused with proximal tubular equilibrium solutions containing radioactive D-glucose. The transport rates follow 2-parameter kinetics: Km = 5.7 mmol/I. Vmax - 145 X 10(-12) MOL/MIN-1; Phlorrhizin inhibits transport almost completely, indicating little passive diffusion of D-glucose. The glucose transport capacity in the short loops of Henle, presumably in the pars rects, is much smaller than that in superficial segments.

Animals↗

Micropuncture study of water, electrolytes, and urea movements along the loops of henle in psammomys.

The mechanism by which the osmotic pressure increases in tubular fluid along the descending limb of the loop of Henle was examined in Psammomys undergoing salt diuresis. In two series of experiments, micropuncture samples were collected either from proximal and distal convolutions at the surface of the cortex, or from loops of Henle and collecting ducts at the surface of the extrarenal part of the papilla. Inulin-(3)H, urea-(14)C, Na(+), and K(+) concentrations, as well as osmotic pressure, were determined in all micropuncture samples.Net movements of water along the descending and ascending limbs of the loop could not be deduced by comparing inulin data obtained from convoluted tubules and from loops of Henle, since there appeared to be a large difference in the filtration rate of the superficial glomeruli (9 nl/min) and the deep ones (21.4 nl/min) under the conditions of these experiments. The results indicate that no large net movement of water occurred along the loop since a) only 23% of the filtrate was reabsorbed along the loop of Henle (including pars recta) of superficial nephrons despite the fact that all these loops reached markedly hypertonic regions; b) there was no positive correlation between (F/P)(In) in early distal samples and the simultaneous osmotic pressure of the urine; c) when (F/P)(In) and (F/P)(Osm) in loop samples were correlated, the increase in inulin concentration accounted only for 15% of the increase in osmotic pressure. Therefore, 85% of the concentrating process taking place along the descending limb must have resulted from net addition of solutes; this was directly supported by Na(+) and K(+) measurements in the loop samples, which showed that, at the tip of the loops, the Na(+) and K(+) flow rates were correlated to the osmotic pressure. Moreover, since the load of Na(+) urea flow rate in superficial early distal tubules was constant and independent of the urinary osmotic pressure, it is suggested that a medullary recycling of both ions occurred between ascending and descending limbs.Urea-(14)C concentration in the loop samples indicates a net addition of urea into the descending limb; the mean and K(+) delivered to the distal superficial tubules was 4.18 times its filtration rate, suggesting a recycling of urea from collecting ducts to Henle's loops. The permeability properties of the wall of the thin descending limb are discussed in relation to the obtained results.

Animals↗

Thick ascending tubular cells in the loop of Henle: regulation of electrolyte homeostasis.

Renal medullary tubular cells in the loop of Henle have crucial importance for the regulation of homeostasis of the extracellular fluid. These cells receive limited amount of blood and oxygen, and are also constantly challenged by the hypertonic environment. The medullary tubular cells in the last part of the loop of Henle have one of the highest known contents of mitochondria of all mammalian cells, reflecting their need for oxidative metabolism in order to sustain high ATP production for active transepithelial electrolyte transport. The commonly used diureticum furosemide targets one of the transporters present in these tubular cells with resulting diuresis. Several pathological states are associated with altered function of the medullary tubular cells, and the nephrotoxic substances tacrolimus and cyclosporine act on these cells. The specific Tamm-Horsfall glycoprotein is produced by medullary tubular cells. Alteration in the urinary excretion of this protein is used as marker of tubular damage.

Animals↗

Physical factors influencing fluid reabsorption from Henle's loop.

Our objective was to produce reductions in the luminal volume of Henle's loop and increases in linear flow velocity through the loop. We did this in a recollection micropuncture study by collecting fluid with and without suction from early distal tubules. With suction, transit time of fast green dye through the loop decreased by 34%, calculated loop volume decreased by 28%, and fractional water reabsorption fell from 73.6 to 70.3% (p smaller than 0.025) in water diuretic rats. Absolute water reabsorption did not decrease significantly. In urea-saline dieuretic rats transit time decreased 25%, calculated loop volume decreased 22%, fractional reabsorption fell from 59.0 to 51.7% (smaller than 0.001), and absolute reabsorption decreased by 2.3 nl/min (p smaller than 0.025). Single nephron glomerular filtration rate, distal tubular sodium concentration, and osmolality were unaffected. The less pronounced effect of collection with suction in water diuretic rats may be related to the lower medullary fluid osmolality, which was 338 plus or minus 9 (S.E.) mOsmol/kg as compared to 497 plus or minus 35 in urea saline diuretic rats. Collecting fluid with suction from late proximal tubules did not alter glomerular filtration rate or fractional water reabsorption. Stumpe et al. ((1970) J. Clin. Invest. 49, 1200-1212) noted an inverse correlation between fluid reabsorption from Henle's loop and flow velocity in rats with hypertension or congestive heart failure. One can reproduce this correlation by artificially altering the transmural pressure gradient in the loop.

Animals↗

Renin secretion and loop of Henle chloride reabsorption in the adrenalectomized rat.

Renin release is increased in the adrenalectomized rat and is not inhibited by sodium chloride administration. The purpose of this study was to determine whether increased renin release is related to impaired absorptive chloride transport in the loop of Henle. Chloride transport in the loop was measured before and after acute saline infusion in three groups of rats: 1) saline-drinking adrenalectomized rats (Adx); 2) saline-drinking dexamethasone-treated adrenalectomized rats (Dex); and 3) water-drinking sham-operated controls. Unrelated to differences of arterial pressure, glomerular filtration rate, or net sodium chloride balance, chloride reabsorption in the loop of Henle of Adx [836 +/- 172 peq/min (SE)] was less (P less than 0.01) than in controls (1,646 +/- 353) and Dex (1,377 +/- 318) before saline infusion. After saline infusion, chloride delivery to the loop increased (P less than 0.05) in all three groups. However, loop chloride reabsorption increased (P less than 0.01) only in controls and Dex but not in Adx. Before saline infusion, plasma renin concentration (PRC) of Adx (350 +/- 108 U/ml) was greater (P less than 0.01) than that in controls (56 +/- 6) or Dex (108 +/- 36); sodium chloride infusion failed to inhibit PRC in Adx, whereas PRC was suppressed (P less than 0.01) by saline in Dex and controls. Thus stimulation of renin release in adrenalectomized animals was associated with decreased absorptive chloride transport in the loop of Henle. Dexamethasone normalized loop function and renin responsiveness to sodium chloride.

Absorption↗

Histotopography and ultrastructure of the thin limbs of the loop of Henle in the hamster.

In the kidney of the Syrian hamster the descending thin limbs of both the short and long loops of Henle are not spatially separated from each other and descend between the vascular bundles. Ultrastructurally, five different epithelial types are distinguished in the thin limbs of the short and long loops of Henle. Short loops possess only a descending thin limb with a simply organized epithelium (type 1). Long loops comprise an upper and a lower part of the descending thin limb and the ascending thin limb. The upper part of the long descending thin limb is equipped with a complex and highly interdigitating epithelium with shallow junctions (type 2), which gradually transforms into the simple noninterdigitating type-3 epithelium of the lower part. In a minor portion of long descending thin limbs, however, the upper part begins with an even more complexly organized epithelium (type 2a) than type 2. Type-2a epithelium is conspicuously thicker and possesses a more elaborate mode of cellular interdigitation. Along the descent of this tubular part through the inner stripe of the outer medulla, type-2a epithelium transforms into type-2 epithelium. It is suggested that the long descending thin limbs, which start with type-2a epithelium, belong to the longest loops. The type-4 epithelium of the ascending thin limbs is characterized by flat and extensively interdigitating cells with shallow junctions. The unique pattern of the type-2a epithelium favors the assumption that solute secretion essentially contributes to the increase in concentration of tubular fluid in long descending thin limbs.

Animals↗

Basic amino acid transport in renal papilla: microinfusion of Henle's loops and vasa recta.

To determine whether basic amino acids, like acidic and neutral amino acids, could be reabsorbed distal to tips of Henle's loops and recycled between loops and vasa recta in the renal papilla, we continuously microinfused ascending Henle's loops and vasa recta with 14C-labeled L-lysine (L-Lys; 1.28 mM) or L-arginine (L-Arg; 1.17 mM) and 3H-labeled inulin. We also determined percent of recovered radiolabel as intact amino acid. Like acidic and neutral amino acids, relative to inulin, approximately 30% of L-Lys and approximately 45% of L-Arg microinfused into Henle's loops were reabsorbed. However, whereas radiolabeled L-Lys reabsorption, like reabsorption of acidic and neutral amino acids, was not readily inhibited, radiolabeled L-Arg reabsorption was reduced to approximately 25% by addition of unlabled L-Arg (50 mM) or L-homoarginine (L-Homo-Arg) (50 mM) to infusate. This observation provides greater evidence for specific, carrier-mediated reabsorption for L-Arg than for acidic or neutral amino acids. About 36% (relative to inulin) of each of these amino acids microinfused into ascending vasa recta apparently was transferred directly into ipsilateral tubular structures (probably thin descending limbs of Henle's loops). Transfer of radiolabeled L-Arg was reduced to approximately 8% by the inclusion of unlabeled L-Arg (50 mM) in infusate. Transfer of unlabeled L-Lys was unaffected by inclusion of unlabeled L-Lys (50 mM) in infusate but was reduced to approximately 20% by inclusion of unlabeled L-Arg (50 mM) or L-Homo-Arg (50 mM) in infusate. (ABSTRACT TRUNCATED AT 250 WORDS)

Amino Acids↗

Consequences of potassium recycling in the renal medulla. Effects of ion transport by the medullary thick ascending limb of Henle's loop.

The consequences of K recycling and accumulation in the renal medulla were examined by measuring the effect of elevated K concentration on ion transport by the medullary thick ascending limb of Henle's loop. Perfused and bathed in vitro, thick limbs from both mouse and rabbit displayed a graded, reversible reduction of transepithelial voltage after increasing K concentration from 5 to 10, 15, or 25 mM. The effect was reproducible whether osmolality was 328 or 445 mosmol/kg H2O, and whether K replaced Na or choline. Net chloride absorption and transepithelial voltage were reduced by almost 90% when ambient K concentration was 25 mM. When either lumen or bath K was increased to 25 mM, net Na absorption was reduced. There was spontaneous net K absorption when perfusate and bath K concentration was 5 mM. Analysis of transepithelial K transfer after imposition of chemical gradients demonstrated rectification in the absorptive direction. Absorption of K by this segment provides a means to maintain high medullary interstitial concentration. Accumulation of K in the outer medulla, by reducing NaCl absorption, would increase volume flow through the loop of Henle and increase Na and water delivery to the distal nephron. K recycling thus might provide optimum conditions for K secretion by the distal nephron.

Absorption↗

Regulation by adrenal corticosteroids of sodium and potassium transport in loop of Henle and distal tubule of rat kidney.

Studies were conducted to examine the effects of adrenalectomy (ADX) and selective, physiological adrenal corticosteroid replacement on sodium and potassium transport by the superficial loop of Henle and distal tubule of rat kidney in vivo. In the loop of Henle, ADX inhibited sodium reabsorption by 33%. Whereas dexamethasone had no effect on reabsorption, aldosterone increased sodium transport to control levels. Thus, physiological levels of mineralocorticoids, but not glucocorticoids, control a fraction of sodium reabsorption in the loop of Henle. ADX also inhibited potassium reabsorption in the loop of Henle. Both dexamethasone and aldosterone reversed the inhibition, although only aldosterone increased reabsorption to control levels. In the distal tubule, ADX reduced sodium reabsorption by 44%. Both aldosterone and dexamethasone stimulated reabsorption: however, only aldosterone increased transport to control. Potassium secretion by the distal tubule was also reduced 34% by ADX. Aldosterone, but not dexamethasone, stimulated secretion. Thus, physiological levels of aldosterone regulate a fraction of sodium reabsorption and potassium secretion in the distal tubule.

Absorption↗

Role of Ca2+-activated K+ channel in regulation of NaCl reabsorption in thick ascending limb of Henle's loop.

1. Reabsorption of NaCl in the thick ascending limb of Henle's loop involves the integrated function of the Na+,K+,Cl- -cotransport system and a Ca2+-activated K+ channel in the luminal membrane with the Na+,K+-pump and a net Cl- conductance in the basolateral membrane. 2. Assay of K+ channel activity after reconstitution into phospholipid vesicles shows that the K+ channel is stimulated by Ca2+ in physiological concentrations and that its activity is regulated by calmodulin and phosphorylation from cAMP dependent protein kinase. 3. For purification luminal plasma membrane vesicles are isolated and solubilized in CHAPS. K+ channel protein is isolated by affinity chromatography on calmodulin columns. The purified protein has high Ca2+-activated K+ channel activity after reconstitution into vesicles. 4. The purified K+ channel consists of two proteins of 51 and 36 kDa. Phosphorylation from cAMP dependent protein kinase stimulates K+ channel activity and labels the 51 kDa band. The 36 kDa band is rapidly cleaved by trypsin and may be involved in Ca2+ stimulation. 5. Opening of the K+ channel by Ca2+ in physiological concentrations and regulation by calmodulin and phosphorylation by protein kinase may mediate kinetic and hormonal regulation of NaCl transport across the tubule cells in TAL.

Animals↗

Proteomic analysis of cellular response to osmotic stress in thick ascending limb of Henle's loop (TALH) cells.

Epithelial cells of the thick ascending limb of Henle's loop (TALH cells) play a major role in the urinary concentrating mechanism. They are normally exposed to variable and often very high osmotic stress, which is particularly due to high sodium and chloride reabsorption and very low water permeability of the luminal membrane. It is already established that elevation of the activity of aldose reductase and hence an increase in intracellular sorbitol are indispensable for the osmotic adaptation and stability of the TALH cells. To identify new molecular factors potentially associated with the osmotic stress-resistant phenotype in kidney cells, TALH cells exhibiting low or high levels of resistance to osmotic stress were characterized using proteomic tools. Two-dimensional gel analysis showed a total number of 40 proteins that were differentially expressed in TALH cells under osmotic stress. Twenty-five proteins were overexpressed, whereas 15 proteins showed a down-regulation. Besides the sorbitol pathway enzyme aldose reductase, whose expression was 15 times increased, many other metabolic enzymes like glutathione S-transferase, malate dehydrogenase, lactate dehydrogenase, alpha enolase, glyceraldehyde-3-phosphate dehydrogenase, and triose-phosphate isomerase were up-regulated. Among the cytoskeleton proteins and cytoskeleton-associated proteins vimentin, cytokeratin, tropomyosin 4, and annexins I, II, and V were up-regulated, whereas tubulin and tropomyosins 1, 2, and 3 were down-regulated. The heat shock proteins alpha-crystallin chain B, HSP70, and HSP90 were found to be overexpressed. In contrast to the results in oxidative stress the endoplasmic reticulum stress proteins like glucose-regulated proteins (GRP78, GRP94, and GRP96), calreticulin, and protein-disulfide isomerase were down-regulated under hypertonic stress.

Animals↗

Effect of loop of Henle flow rate on glomerular capillary pressure.

Studies were performed in male Munich-Wistar rats weighing a mean of 100 g to evaluate further the hemodynamic effects of changing loop of Henle flow rate. When late proximal flow rate was varied by microperfusion from the late proximal tubule, single nephron glomerular filtration rate (SNGFR) measured in the early proximal tubule fell from 17.2 +/- 0.86 nl min-1 at a perfusion rate of zero to 11.8 +/- 0.88 nl min-1 at a perfusion rate of 14 nl min-1. Further increases in loop flow did not further depress SNGFR. Glomerular capillary pressure (Pgc) was measured directly in superficial glomeruli while loop of Henle flow was varied. Elevating loop flow was followed by a rapid decrease in Pgc, which was reversible when flow was reduced. The mean response to maximum stimulation was a decrease in Pgc of 6.6 +/- 0.7 mm Hg, and the response was close to maximum at a perfusion rate of 14 nl min-1. Parallel changes in stop-flow pressure were observed. These findings, together with previous observations that increasing loop of Henle flow results in decreased single nephron blood flow, permit the conclusion that the dominant vascular response to elevated loop of Henle flow is constriction of the afferent arteriole.

Animals↗

Adenosinetriphosphatase activity in the cell membranes of kidney tubule cells.

This cytochemical study demonstrates high levels of apparent ATPase activity in the infolded cell membranes of the proximal tubules (dog, rat, human, mouse, monkey, and opossum) and ascending loops of Henle (dog, rat, human and, to a variable degree, mouse). Electron microscopy has shown (see Rhodin (1)) that these membranes separate adjacent cells where they interlock with one another by multiple cytoplasmic lamellae containing oriented mitochondria. The significance of the high ATPase activity is considered in relation to possible movements of the membranes and to "active transport" believed to occur there. In the rat, enzyme activity in the proximal tubule membranes does not survive formol-calcium fixation, and it is therefore necessary to use unfixed sections for its demonstration. However, in edematous rats with experimental nephrosis (induced by the injection of aminonucleoside or with antikidney serum) marked ATPase activity is exhibited in these membranes even after formol-calcium fixation. When proximal tubule or Henle loop cells of the dog acquire an altered metabolism, as indicated by accumulated lipide spheres or by "droplets," the infolded ATPase-rich membranes are no longer demonstrable.

Adenosine Triphosphatases↗

Kidney in the aging cat: hexokinase, aldolase, L-alpha-glycerophosphate dehydrogenase, and lactate dehydrogenase histochemistry.

Hexokinase, aldolase, L-alpha-glycerophosphate dehydrogenase, and lactate dehydrogenase activities were determined in the kidney of the aging cat. Kidneys from 24 domestic cats 2 months to 7.5 years old in 6 age groups were examined by light microscopic and histochemical methods. Enzyme activities in anatomic components of the kidney were assessed on a quantitative basis for evaluation of mean activity between the age groups. In the cats with advancing age, renal components generally had stable activity. A significant (P less than 0.05) increase in hexokinase activity occurred with advancing age in the ascending part of the renal loop (Henle's loop) and in the distal convoluted tubule. Significant (P less than 0.05) increases in aldolase activity with aging were in cortical connective tissue, internal part of the glomerular capsule (podocytes), distal convoluted tubule, and convoluted segment (Pi) of the proximal portion of the nephron tubule. L-alpha-glycerophosphate dehydrogenase and lactate dehydrogenase activity increased significantly with aging in the convoluted (Pi) segment of the proximal portion of the nephron tubule.

Aging↗

Does defective chloride reabsorption at the loop of Henle play a major role in the pathogenesis of Bartter's syndrome?

In a patient with typical features of Bartter's syndrome, intrarenal reabsorption of sodium and water at different sites of the nephron, during maximal water diuresis, was studied twice in the course of the disease. During the first study, chloride reabsorption at the loop of Henle and renal diluting ability were not impaired. Chloride reabsorption at the loop of Henle was 0.72 (normal values 0.66-0.90), minimal urine osmolality 71 mosm/kg (normal values less than 75 mosm/kg) and chloride fractional clearance 0.44 (normal values 0.3-6.8). An impaired chloride reabsorption at the loop of Henle was found 6 months later. Chloride reabsorption at the loop of Henle was 0.28, minimal urine osmolality 197 mosm/kg and chloride fractional clearance 8.23. These data suggest that the defect in chloride reabsorption at the loop of Henle is not a pathophysiological abnormality of the syndrome because it appeared at a later time than the other symptoms.

Absorption↗

Periodic disklike structures of the adult human loop of Henle. Light and electron microscopic studies.

A total of 51 adult human kidneys was examined for the presence of the periodic disklike thickenings recently described by Belliveau in the basement membrane of the loop of Henle. These "Belliveau bodies" formed a PAS-positive rectangular lattice within the basement membrane and appeared limited to the loop of Henle. The bodies were seen in the medulla of every kidney examined but varied considerably in the frequency of their appearance from case to case. This variability did not correlate with predominant illness, organ weight, or time interval between death and autopsy. They averaged 9 to 14 micron in diameter with a horizontal and vertical periodicity of 12 and 16 micron, respectively. Although the bodies were PAS positive and diastase resistant, they did not stain with Congo red, mucicarmine, reticulin, or elastin staining techniques. Ultrastructurally, they consisted of multilaminated areas of basement membrane material. The constancy of their appearance in the material examined suggests that they are a normal anatomic feature or possibly an age-related change of the basement membrane of Henle's loop in the human kidney.

Adult↗

Axial heterogeneity of potassium transport across hamster thick ascending limb of Henle's loop.

Functional significance of morphological heterogeneities along the thick ascending limb of Henle's loop of hamsters was explored by the in vitro microperfusion technique with special reference to K+ transport. The transmission electron microscopic study confirmed that there are two types of cells, with smooth surface (S-cell) and rough surface (R-cell), respectively, and that the former is abundant in the medullary thick ascending limb (MTAL), whereas the latter is in the cortical portion (CTAL). The electrophysiological study revealed that in both segments there are two cell populations, one having high basolateral and low apical membrane K+ conductances (HBC) and the other having low basolateral and high apical K+ conductances (LBC). Random cell puncture revealed that the ratios of HBC/LBC were 24/7 (77%/23%) in the MTAL and 7/22 (24%/76%) in the CTAL, suggesting that HBC corresponds to S-cell, whereas LBC corresponds to R-cell. Net K+ transport was determined in two segments by measuring K+ concentration in the collected and perfused fluid by ultramicroflame photometry. In all six tubules of MTAL, net K+ flux had a direction to reabsorption with a mean of 4.87 +/- 0.46 pmol.min-1.mm-1. In marked contrast, in all six tubules of CTAL, we observed K+ secretion with a mean of -3.81 +/- 0.49 pmol.min-1.mm-1. The transmural voltage was positive in both segments and was significantly higher in the CTAL (7.8 +/- 0.5 mV) than in the MTAL (2.5 +/- 0.2 mV). From these observations, we conclude that the S-cell corresponding to the HBC cell reabsorbs K+, whereas the R-cell corresponding to the LBC cell secrets K+.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗