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  • 1
    Electronic Resource
    Electronic Resource
    Springer
    Journal of molecular medicine 60 (1982), S. 1103-1113 
    ISSN: 1432-1440
    Keywords: Proximal tubule ; Thick ascending limb ; Henle's loop ; Membrane transport ; Transport alteration ; Diuretics ; Proximaler Tubulus ; dicker aufsteigender Teil ; Henle'sche Schleife ; Membrantransport ; Transportstörung ; Diuretika
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Description / Table of Contents: Zusammenfassung Die Kenntnisse der renalen Transportmechanismen sind während den letzten Jahren dank den Fortschritten in der Methodik enorm angewachsen. Der transepitheliale Transport sowie seine pathologische und pharmakologische Veränderung können teilweise als Membranphänomene verstanden werden. Am Beispiel natriumgekoppelter Transportmechanismen im proximalen Tubulus sowie des Natrium-chlorid-Transportes in der dicken aufsteigenden Henle'schen Schleife werden in diesem Artikel Membrantransportmechanismen diskutiert. Es wird gezeigt, daß geänderte Nierenfunktion oft mit geändertem Membrantransport gleichzusetzen ist.
    Notes: Summary Over the past few years, our knowledge on renal tubular transport mechanisms has increased considerably. Due to new technical developments, it is now possible to understand in part transepithelial transport and its pathological and pharmacological alterations at the level of the cell membranes. Different membrane transport mechanisms are discussed in this article, whereby sodium coupled solute transport in the proximal tubule and sodium chloride transport in the thick ascending limb of Henle's loop are taken as examples. It is indicated that an altered function of the kidney can often be equated with an alteration of the membrane transport.
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    Springer
    Journal of molecular medicine 61 (1983), S. 1019-1027 
    ISSN: 1432-1440
    Keywords: Loop diuretics ; Thick ascending limb ; Henle's loop ; Diluting segment ; Na+ - 2Cl− ; K+-cotransport
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary During the passed few years the cellular mechanisms responsible for the NaCl reabsorption in the thick ascending limb of the Henle loop of mammalian nephron and of the early distal tubule of amphibian kidney have been extensively studied. From these studies a new type of secondarily active transport mechanism, i.e. the Na+ - 2Cl− - K+ symport has emerged. Meanwhile it has been recognized that this system is also present in many other epithelia. All these epithelia share in common that they are sensitive to the so called loop diuretics. The present article summarizes our current knowledge of how the loop diuretics, by reversible interaction with the above cotransport system, inhibit the NaCl reabsorption in the thick ascending limb of Henle's loop. It is shown that these drugs transfer the thick ascending limb cell to a state in which not only transepithelial NaCl reabsorption ceases but in which also very little energy is consumed since then K+ and Cl− “relax” to passive distribution across both cell membranes and Na+ entry into the cell is blocked.
    Type of Medium: Electronic Resource
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  • 3
    Electronic Resource
    Electronic Resource
    Springer
    Pflügers Archiv 390 (1981), S. 30-37 
    ISSN: 1432-2013
    Keywords: Rabbit kidney ; Thick ascending limb of Henle's loop ; Paracellular shunt ; Furosemide ; Ouabain
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract The cation permeability of the cortical thick ascending limb of Henle's loop was investigated with electrophysiological methods in the isolated perfused tubule preparation of rabbit kidney. The transepithelial specific resistance (R T) and electrical potential difference (PD) were determined in 4 experimental groups. In group 1 (n=51) the tubules were perfused with a modified Ringer's solution on both sides of the epithelium; the PD was 7±0.4 mV lumen positive, and theR T 34±2 Ω·cm2. In group 2 (n=12) one of both sides of the epithelium was perfused with dilute (54 mmol/l) NaCl solutions under control conditions and in the absence of active transport. Inhibition was obtained in four different ways: low temperature (22° C), zero K+ solutions on both sides of the epithelium, 5·10−5 mol/l furosemide, added to lumen perfusate, or 10−5 mol/l ouabain added to the bathing solution. In the presence and in the absence of active transport a NaCl gradient of 154 versus 54 mmol/l induced diffusion potentials across the epithelium which were symmetrical and of nearly equal magnitude: +12, −14 and +15, −14 mV respectively. In group 3 (n=51) Na+ was completely replaced by choline+, tetraethylammonium+, tris-hydroxymethyl-aminomethane+, or Li+ in either bath or lumen perfusate or in both perfusates. The biionic diffusion potentials were symmetric; the replacement of Na+ by these cations on both sides markedly increasedR T. Both kinds of measurements yielded a permeability sequence ofP Na +〉P Li +〉P organic cation. In group 4 (n=17) 50 mmol/l of Na+ was replaced by K+, Li+, Rb+, or Cs+ on one of the sides and active transport was inhibited by furosemide or ouabain. From the membrane diffusion potentials and theR T values in group 4 as well as in group 3 the following cation permeability sequence was calculatedP K +〉P Na +〉P Rb + =P Li +〉P Cs +〉P organic cation. It is concluded that the cortical thick ascending limb of Henle's loop has a low resistance pathway which is cation selective similar to that of “leaky” epithelia. Since the membrane diffusion potentials are symmetric and since they are not altered by inhibition of active transport, it is likely that this low resistance pathway is formed by a paracellular shunt.
    Type of Medium: Electronic Resource
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  • 4
    ISSN: 1432-2013
    Keywords: Amphibian kidney ; Potassium ; Chloride ; Barium ; Furosemide
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Experiments in the amphibian early distal tubule have shown that Cl transport is secondarily active, coupled to the flux of Na and dependent on the presence of luminal K. Omission of luminal K results in a decrease of cellular Na, a finding that suggests inhibition of luminal Na entry. In the present study intracellular chloride activity (Cli) and peritubular cell membrane potentials (PDpt) were evaluated before and after omission of luminal K. Furthermore, the effect of inhibition of the luminal K conductance by barium on the electrochemical gradient of Cl (E te Cl ) across the distal epithelium was determined at static head conditions. Experiments were performed in early distal segments of the isolated perfused kidney ofAmphiuma andRana esculenta. Cli and PDpt were measured simultaneously in single cells by double barreled Cl sensitive microelectrodes in the presence and absence of luminal K. E te Cl was determined at zero net flux conditions with single barreled electrodes in control tubules, in the presence of barium (3·10−3 mol/l) and in the presence of furosemide (5·10−5 mol/l). In 26 individual cellular impalements omission of luminal K hyperpolarized PDpt from 72.5±1.2 to 90.0±1.9 mV (cell interior negative). Concomitantly, Cli fell from 8.5±0.4 to 5.4±0.3 mmol/l. Both effects occurred within seconds and were fully reversible. Addition of barium to the luminal fluid diminished E te Cl (directed lumen positive) from a control value of 39.5±1.4 mV to 28.5±2.5mV. E te Cl could be further diminished to 14.1±2.1 mV and to 1.3±0.5 mV after application of barium on both sides and after luminal application of furosemide, respectively. The experiments indicate that active Cl uptake across the luminal cell membrane depends critically on the presence of luminal K. Omission of luminal K achieved either by perfusing the lumen with K-free solutions or by inhibition of K back flux from the cell interior into the lumen by barium reduces Cl reabsorption. Together with previous data on the K dependence of the Na uptake the present experiments support the hypothesis of a common transport system for K, Na, Cl located in the luminal cell membrane.
    Type of Medium: Electronic Resource
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  • 5
    ISSN: 1432-2013
    Keywords: Isolated perfused tubule ; Rabbit cortical thick ascending limb of Henle's loop ; Sodium potassium-chloride cotransport ; Ba2+
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Previous data from our laboratory have shown that active transport in the cortical thick ascending limb of Henle's loop (cTAL), as measured by the short circuit current (ISC, μA · cm−2), requires the presence of Na+ and Cl−. The data were compatible with the model of secondarily active Cl− reabsorption involving the cotransport of Na+ and Cl− across the luminal membrane. The data suggested, furthermore, that 1 Na+ and 2 Cl− interact with the luminal carrier. In the present study it was tested whether this reabsorptive mechanism also requires the presence of luminal K+. Isolated cTAL segments (n=40) were perfused at high flow rates with a modified Ringer's solution. Removal of K+ from the lumen reduced ISC significantly from 215 to 133 μA·cm−2. Addition of Ba2+ (10−3 mol·l−1) which blocks the K+ conductance of the luminal membrane, to the K+-containing lumen perfusate decreased ISC significantly from 234 to 141 μA·cm−2. Combination of both manoeuvres: perfusion with a K+-free and Ba2+-containing solution almost abolished ISC from a control of 237 to 56 μA · cm−2. The results are compatible with the view that in rabbit cTAL the luminal carrier interacts with all 3 ions, possibly 1 Na+, 2 Cl−, and 1 K+. K+ recycles across the luminal membrane through its conductive pathway.
    Type of Medium: Electronic Resource
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  • 6
    Electronic Resource
    Electronic Resource
    Springer
    Pflügers Archiv 401 (1984), S. 424-426 
    ISSN: 1432-2013
    Keywords: Proximal tubule ; patch-clamp ; K+-channels
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract A new method is described, which enables the recording of single ionic channels from the basolateral as well as the luminal membrane of renal proximal tubules with the patch-clamp technique. Segments of late proximal tubules of rabbit kidney are cannulated and perfused from one end. The other end is open and freely accessible to a patch pipette. The patch electrode can be moved against lateral cell membranes or can be inserted through the open end into the lumen and brought to contact with the brush-border membrane. Both, in the basolateral as well as in the luminal membrane, giga seals can be achieved. In both membranes, K+ selective channels could be identified.
    Type of Medium: Electronic Resource
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  • 7
    ISSN: 1432-2013
    Keywords: Pseudomonas aeruginosa toxin ; thick ascending limb of the loop of Henle ; rabbit
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract The present study examines directly the effect of a cytotoxin of Pseudomonas aeruginosa on the in vitro perfused rabbit cortical thick ascending limb of the loop of Henle (cTAL). 25 cTAL segments were perfused at high rate. The open circuit transepithelial electrical PD (PDte) and the specific electrical transepithelial resistance (Rt) were recorded continuously. From PDte/Rt the equivalent short circuit current (Isc) was calculated. The Isc was 214±30 μA·cm−2 under control conditions, and decreased significantly to 74±34 μA·cm−2 60 s after the addition of toxin (2 mg·l−1) to the lumen perfusate. Microscopic observation and photographs taken at that time clearly indicated swelling of the cTAL cells. Thereafter inhibition of active transport proceeded further, Rt fell progressively, and cells started to desquamate from the basement membrane. This effect of the toxin was dose dependent, and was half maximal at approximately 1.2 mg·l−1. From the bath side the effect was less marked and higher doses of toxin had to be used (half maximal effect at 5 mg·l−1). We conclude that this toxin of Pseudomonas aeruginosa exerts its toxic effect on the cTAL segment by increasing primarily the permeability of the lumen membrane.
    Type of Medium: Electronic Resource
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  • 8
    ISSN: 1432-2013
    Keywords: Intracellular sodium activity ; Early distal tubule ; Sodium chloride cotransport ; Potassium ; Sodium sensitive microelectrode
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract From previous studies it is known that a furosemide-sensitive sodium chloride cotransport system is operative in the luminal cell membrane of the early distal amphibian tubule. Since inhibition of sodium chloride cotransport prevents potassium reabsorption in this nephron segment, experiments were carried out to evaluate further the possible relationship between sodium chloride and potassium transport by studying the changes of cellular sodium activity following luminal deletion of potassium ions. Sodium-sensitive liquid ion exchange microelectrodes and conventional microelectrodes were employed to determine the transpithelial potential (PDte), the peritubular cell membrane potential (PDpt) and the intracellular sodium activity (Nai +) in the presence and absence of luminal potassium. The ratio of the luminal cell membrane resistance over the peritubular cell membrane resistance (Rlu/Rpt) was also estimated. When potassium ions are omitted from the luminal perfusate, PDpt hyperpolarizes by some 20 mV, PDte approaches zero and Nai + decreases by about 40%. Rlu/Rpt is more than doubled in the presence of a potassium-free perfusate. Both potential and resistance changes are fully reversible. Similar results were obtained in experiments in which Barium ions (1 mmol/l BaCl2) were present during the luminal potassium substitution. Our results indicate that absence of potassium inhibits luminal sodium chloride entry; as a result of continued peritubular sodium extrusion cellular sodium activity falls. The increase of Rlu/Rpt following perfusion with a potassium-free perfusate is interpreted as a decrease of a significant electrodiffusive potassium conductance in the luminal cell membrane.
    Type of Medium: Electronic Resource
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  • 9
    Electronic Resource
    Electronic Resource
    Springer
    Pflügers Archiv 389 (1981), S. 175-176 
    ISSN: 1432-2013
    Keywords: Isolated renal tubule ; in vitro perfusion
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract A modified system for the in vitro perfusion of isolated tubule segments is presented. The system consists of four holders each of which carries an acrylic cylinder. The acrylic cylinders are used to fix the glass pipettes in a concentric position. The four holders are mounted onto a support consisting of two holding pieces and three steel rods. Three of the holders contain ball-races so that they can slide on the rods with high accuracy and little friction. The holers to which the sylgard pipette and the perfusion pipette are attached are moved by electric motors. Compared with the classical V-track system this modification has the advantage of higher precision. Once the different pipettes are centered, concentricity is maintained even when the pipettes are moved forward or backward. Thus, this equipment facilitates the cannulation of tubules and increases the number of successfull experiments.
    Type of Medium: Electronic Resource
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  • 10
    Electronic Resource
    Electronic Resource
    Springer
    Pflügers Archiv 395 (1982), S. 121-125 
    ISSN: 1432-2013
    Keywords: Bicarbonate ; Renal tubular transport ; Carbonic anhydrase inhibition ; Permeability ; Microperfusion ; Micropuncture
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract The present study was designed to define the prerequisites of carbonic anhydrase independent bicarbonate reabsorption. In free flow experiments during systemic application of carbonic anhydrase inhibitor benzolamide (50 mg/kg B. W.) bicarbonate recovery in % of filtered load was found to be 74±8% in late proximal convoluted tubules, 39±6% in distal convoluted tubules and 32±4% in urine, indicating that most of carbonic anhydrase independent bicarbonate reabsorption occurs in tubule segments prior to distal convoluted tubules. In vivo continuous microperfusion experiments in proximal convoluted tubules demonstrated that luminal benzolamide (0.5 mmol/l) virtually abolishes net bicarbonate fluxes, when bicarbonate concentration in the luminal perfusate (25 mmol/l) is close to peritubular plasma concentration (24.4 mmol/l). In contrast, a significant downhill reabsorptive flux occurs, when perfusate bicarbonate concentration is 75 mmol/l and a significant downhill secretory flux is observed, when the perfusate is initially free of bicarbonate. The corresponding apparent permeabilities are 1.0±0.1·10−6 cm2/s for influx and 1.6±0.4·10−6 cm2/s for efflux of bicarbonate. Clearance studies reveal that carbonic anhydrase dependent and independent bicarbonate reabsorption are not saturable but depend on the rate of volume reabsorption in the kidney. In conclusion, passive movements of bicarbonate do occur in proximal convoluted tubules and most likely contribute to carbonic anhydrase independent bicarbonate reabsorption.
    Type of Medium: Electronic Resource
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