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  • 1
    Electronic Resource
    Electronic Resource
    Springer
    Pflügers Archiv 368 (1977), S. 245-252 
    ISSN: 1432-2013
    Keywords: Renal tubule ; H+ ion secretion ; Na+ coupled transport ; Ouabain ; SITS
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary The rate of active transport by the proximal renal tubule of amino acid (l-histidine), sugar (α-methyl-d-glycoside), H+ ions (glycodiazine), phosphate and para-aminohippurate was evaluated by measuring the zero net flux concentration difference (Δc) of these substances. In the case of calcium the electrochemical potential differenceΔc +zFci Δϕ/RT) was the criterion employed. The rate of isotonic Na+-absorption (JNa) was measured with the shrinking droplet method. The effect of ouabain on the transport of these substances was tested in the golden hamster and the effect of SITS (4-acetamido-4′isothiocyanatostilbene 2,2′-disulfonic acid) was observed in rats. Ouabain (1 mM) applied peritubularly incompletely inhibited JNa (80%), but in combination with acetazolamide (0.2 mM) the inhibition was almost complete (93%). In addition, ouabain inhibited the sodium coupled (secondary active) transport processes ofl-histidine, α-methyl-d-glycoside, calcium and phosphate by more than 75%. It did not affect H+ (glycodiazine) transport and PAH transport was only slightly affected. When SITS (1 mM) was applied from both sides of the cell it inhibited H+ (glycodiazine) transport by 72% and reduced JNa by 38% when given from only the peritubular cell side. SITS (1 mM), however, had no significant affect on H+ secretion and sodium reabsorption if it was applied from only the luminal side. Furthermore it had no affect on the other transport processes tested, regardless of the cell side to which it was applied. When the HCO 3 − buffer or physically related buffers were omitted from the perfusate the absorption of Na+ was reduced by 66%, phosphate by 44%, andl-histidine by 15%. All the other transport processes tested were not significantly affected. The data are consistent with the hypothesis that the active transport processes of histidine, α-methyl-d-glycoside and phosphate, which are located in the brush border, are driven by a sodium gradient which is abolished by ouabain. This may also apply to the Na+-Ca2+ countertransport located at the contraluminal cell side. The residual Na+ transport remaining in the presence of ouabain is likely to be passively driven by the continuing H+ transport which probably is driven directly by ATP. SITS seems to inhibit the exit step of HCO 3 − from the cell and secondary to that, the luminal H+-Na+ exchange and consequently the Na+ reabsorption. In the absence of HCO 3 − buffer in the perfusates the luminal H+-Na+ exchange seems to be affected and the pattern of inhibition of the other transport processes is almost the same as with SITS. The different effects onP i reabsorption observed under these conditions might be explained by possible variations in intracellular pH.
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    Springer
    Pflügers Archiv 357 (1975), S. 149-163 
    ISSN: 1432-2013
    Keywords: Renal Tubule ; H+ Transport ; Sodium Dependence ; Carbonic-Anhydrase Inhibitors ; Adaptation (Acid Base Balance)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Using the stop flow microperfusion technique with simultaneous capillary perfusion the secretory rate of H+ ions in the proximal tubule was evaluated by measuring the level flow reabsorption as well as the static head concentration difference of3H labelled glycodiazine. At ambient glycodiazine concentration of 21 mmol/l the level flow reabsorption is in the same range as that of bicarbonate. In the early proximal loops the reabsorption is 20% greater than in the late proximal loops. The carbonic anhydrase inhibitors acetazolamide and 3,4-methylenedioxyphenyl-sulfonamide (both 10−4 M) as well as furosemide (10−3 M) inhibit the glycodiazine reabsorption 43%, 27% and 22% respectively. Thiocyanate (2 · 10−2 M), however, exerted only an insignificant inhibition (12%). When Na+ in the ambient perfusion solutions was replaced by Li+ or choline+ the glycodiazine transport was strongly reduced. Ouabain (5 · 10−2 M) inhibited too, but amiloride (10−3 M) had no effect on glycodiazine transport. The glycodiazine transport was 28% reduced in metabolic alkalosis and to a smaller although significant extent (17%) in metabolic acidosis; it was unchanged in chronic hypercapnia. In chronic K+ depletion the glycodiazine reabsorption was accelerated by 12% only in the early proximal loops. Chronic parathyroidectomy as well as acute substitution with parathyroid hormone had no effect on the glycodiazine absorption. The main conclusions are: Proximal H+ transport proceeds with suitable buffers. Although independent of HCO3 − and carbonic anhydrase, it could be partially inhibited by CA inhibitors. H+ transport is supposed to proceed as countertransport with Na+ ions. In chronic alkalosis the H+ transport is reduced.
    Type of Medium: Electronic Resource
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  • 3
    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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  • 4
    Electronic Resource
    Electronic Resource
    Springer
    Pflügers Archiv 379 (1979), S. 49-52 
    ISSN: 1432-2013
    Keywords: Renal collecting duct ; Na+ reabsorption ; Adrenalectomy ; Acetazolamide ; Amiloride
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Using the shrinking droplet method and simultaneous perfusion of the peritubular capillaries the isotonic reabsorption of Ringer's solution from the papillary collecting ducts was measured. Under control conditions the volume reabsorption from the papillary collecting ducts wasJ v±SE=2.6±0.1 · 10−5 cm3 · cm−2 · s−1. In rats which were on low Na+ diet,J v increased to 127%, and in adrenalectomized animals it decreased to 34% of the control value. Three hours after application of aldosterone in the adrenalectomized animalsJ v was partially restored to 63% of control rats. Amiloride 10−4 M, added to the luminal perfusate, produced a strong inhibition ofJ v (to 32% of control). Acetazolamide, 10−4 M, added to both perfusates, reducedJ v very strongly (to 40% of control), while omission of bicarbonate reduced it only to 77% of control. Acetazolamide, added to bicarbonate-free perfusates, did not result in a significant further reduction ofJ v. The data indicate that the Na+ reabsorption from the papillary collecting duct is controlled by mineralocorticoids. Furthermore, they suggest the existence of two transport mechanisms in the luminal cell membrane: 1. An amiloride-sensitive entry step and 2. an entry step via a Na+−H+-countertransport mechanism, the latter being less important.
    Type of Medium: Electronic Resource
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  • 5
    Electronic Resource
    Electronic Resource
    Springer
    Pflügers Archiv 389 (1981), S. 271-275 
    ISSN: 1432-2013
    Keywords: Adaptation, HCO 3 − transport ; Glycodiazine transport ; Metabolic acidosis ; Metabolic alkalosis ; Acetazolamide ; SITS ; Potassium deficiency
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Using the technique of capillary perfusion and simultaneous luminal stop flow microperfusion the reabsorption of bicarbonate and glycodiazine from the papillary collecting duct was evaluated. Starting with equal H14CO 3 − and3H-glycodiazine concentrations in the luminal and peritubular perfusates, the decrease in the luminal concentration at 10 and 45 s contact time was measured. In control rats with 25 mmol/l HCO 3 − in the perfusates the rate of HCO 3 − reabsorption calculated from the 10 s values was 0.34 nmol cm−2s−1. In acute metabolic acidosis, the rate of bicarbonate reabsorption was 2,3 times higher. In metabolic alkalosis, the rate of bicarbonate absorption dropped to 13% of the control values. Also the 45 s values of acidotic and alkalotic animals differed significantly from each other. With 25 mmol/l glycodiazine in both perfusates the rate of biffer reabsorption as calculated from the 10 s values was 0.76 nmol cm−2s−1 in control rats and did not deviate significantly from this value in acidotic and alkalotic animals. In control rats the bicarbonate reabsorption in % was the same, no matter whether both luminal and capillary perfusate contained 25 mmol/l bicarbonate or 10 mmol/l. In acidotic rats the rate of HCO 3 − reabsorption did not change significantly if all Na+ in the perfusates was replaced by choline (0.88 versus 0.79 nmol cm−2s−1 at 25 mmol/l HCO 3 − ). When in acidotic rats 0.1 mmol/l acetazolamide or 1 mmol/l SITS (4-acetamido-4′-isothiocyanatostilbene-2,2′-disulfonic acid) was added to both perfusates the rate of HCO 3 − reabsorption dropped by 75 and 58%, respectively. A potassium deficient diet for one week and DOCA administration had no influence on the bicarbonate reabsorption of rats which were on standard diet. The data indicate that (1) the buffer reabsorption from the papillary collecting duct is rather due to H+ ion secretion than to buffer anion reabsorption. (2) The adaptation to metabolic acidosis and alkalosis is specific for bicarbonate and not seen with glycodiazine. (3) Within the concentration range tested the HCO 3 − reabsorption rises linearly with the HCO 3 t- concentration. (4) The HCO 3 − reabsorption in the papillary collecting duct is Na+-independent, it can be inhibited by acetazolamide and SITS, but is not influenced by K+-deficient diet plus DOCA.
    Type of Medium: Electronic Resource
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  • 6
    Electronic Resource
    Electronic Resource
    Springer
    Pflügers Archiv 404 (1985), S. 150-156 
    ISSN: 1432-2013
    Keywords: Glucose ; 2-Deoxy-d-glucose ; Intracellular compartmentalization ; Proximal tubule ; Contraluminal membrane
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract In order to study contraluminal hexose transport, concentration and time-dependent influx of3H-2-deoxy-d-glucose from the interstitium into cortical tubular cells has been measured. The influx curves fit to a two parameter kinetics (K m 1.3±0.2 mmol/l,J max 0.67±0.16 pmol/s · cm) plus an additional diffusion term (withP=6·10−8 cm2/s) and a distribution ratio extracellular to intracellular amount of 2-deoxy-d-glucose of 1∶0.6. Since the extracellular to intracellular free water space as estimated from morphological data was 1∶2, one must conclude that glucose has only free access to 1/3 of the cell water. The intracellularly accessible space was augmented when the tubules were preperfused for 10 s with hypotonic saline. Thereby an increase of the compartment into which diffusion occurs was revealed and a final rupture of this intracellular compartment at 1/4 isotonic solutions was observed. Total replacement of ions in the peritubular perfusate by mannitol did not change 2-deoxy-d-glucose influx, indicating that it is Na+-independent. By adding isotonic concentrations of the respective sugars to the capillary perfusate, three degrees of inhibition of 2-deoxy-d-glucose influx could be revealed: strong inhibition byd-glucose, methyl-β-d-glucoside,d-mannose, 3-O-methyl-d-glucose, 2-deoxy-d-galactose, methyl-β-d-galactoside and 6-deoxy-d-glucose, moderate inhibition byd-galactose,l-glucose,l-mannose andd-fructose, no or borderline inhibition by methyl α-d-glucoside, 2-deoxy-methyl-α-d-galactoside, 1-thio-β-d-glucose, 1-thio-β-d-galactose, 5-thio-α-d-glucose, myo-inositol and mannitol. The contraluminal 2-deoxy-d-glucose influx was also inhibited by phloretin, chlormerodrin and preperfusion with cytochalasin B. Starvation as well as streptozotocin diabetes has no influence on contraluminal 2-deoxy-d-glucose transport. Thus, in contrast to the luminal hexose transport system the contraluminal system is Na+-independent, does not require on OH-group at C-atom 2, acceptsl-glucose and fructose, but not an α-methyl group at C-atom 1.
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  • 7
    Electronic Resource
    Electronic Resource
    Springer
    Pflügers Archiv 407 (1986), S. 488-492 
    ISSN: 1432-2013
    Keywords: Lactate ; Pyruvate ; 3-hydroxybutyrate ; Acetoacetate ; Nonspecific anion channel
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract In order to study the characteristic of contraluminal transport of hydrophylic small fatty acids the in situ stopped flow microperfusion technique [12] has been applied. By measuring with 4 s contact time the decrease in the contraluminal concentration of the respective radiolabelled substances the concentration dependence of the influx into the cortical cells was tested. The 4 s decrease in contraluminal concentration of chloroacetate,l-lactate,d-lactate, 3-hydroxybutyrate and acetoacetate was between 26% and 31%. For each substance the percent decrease was the same, no matter whether it was offered in a concentration of 0.1 or 10 mmol/l. Contraluminal disappearance of 0.1 mmol/ll-lactate was not influenced by 5 mmol/l H2DIDS, probenecid, phloretin, mersalyl or cyanocinnamate, but it was significantly (37%) inhibited by 5-nitro-2-(phenyl-propyl-amino) benzoate, a blocker of the nonspecific anion channel. The percent decrease in propionate uptake was somewhat larger — between 36% and 39% — but again not different at 0.01, 0.1, 1.0 and 10 mmol/l. With pyruvate the contraluminal decrease was 20% at 0.1 mmol/l and 31% at 10 mmol/l. The percent disappearance of the aromatic pyrazinoate was 38% and 34% at 0.1 and 10 mmol/l and for nicotinate 42% and 22%, respectively. The disappearance of nicotinate (0.1 mmol/l) was significantly inhibited by 10 mmol/l pyrazinoate and paraaminohippurate (PAH). The data are in agreement with the hypothesis that the hydrophilic small fatty acids traverse the contraluminal cell side by simple diffusion, possibly via the unspecific anion channel [14], pyruvate via the dicarboxylic acid pathway in a cooperative manner and pyrazinoate, as well as nicotinate, via the PAH pathway.
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  • 8
    Electronic Resource
    Electronic Resource
    Springer
    Pflügers Archiv 405 (1985), S. S106 
    ISSN: 1432-2013
    Keywords: Epithelial transport ; Contraluminal cell membrane ; Dietary adaptation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract In order to study the characteristics of contraluminal phosphate transport the stopped flow microperfusion technique [13] has been applied. By measuring the time-dependent decrease of interstitial33Pi concentration at different starting concentrations a simple diffusion kinetics with a permeability coefficient of 7.5±1.0 · 10−8 cm2 s−1 was found. Such a kinetic was so far only observed with 2-deoxy-d-glucose. This substance, however, is transported in addition by facilitated diffusion as was seen by paraaminohippurate, methylsuccinate and sulfate. The contraluminal transport of phosphate was inhibited by H2-DIDS (5 mmol/l). It was, however, not influenced by omission of Na+ from the perfusates, by addition of sulfate (150 mmol/l), methylsuccinate (50 mmol/l), arsenate (50 mmol/l), the Hg-compound mersalyl (5 mmol/l), high and low phosphate diet and pH changes between 6.0 and 8.0. The data indicate that phosphate, which is reabsorbed from the lumen by a Na+-dependent transport system, leaves the cell by a rather unspecific contraluminal diffusion pathway.
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  • 9
    Electronic Resource
    Electronic Resource
    Springer
    Pflügers Archiv 305 (1969), S. 139-154 
    ISSN: 1432-2013
    Keywords: d-Glucose ; Microperfusion ; Proximal Tubule ; Active Reabsorption ; Kinetic Study ; d-Glucose ; Mikroperfusion ; proximaler Tubulus ; aktive Reabsorption ; kinetische Studien
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Description / Table of Contents: Zusammenfassung Proximale Konvolute von Rattennieren wurden bei fehlendem Nettofluß von Natriumionen und Wasser kontinuierlich mit Lösungen perfundiert, die eined-Glucosekonzentration zwischen 0,5 und 2,0 mmol/l enthielten. Der Abfall der intraluminalend-Glucosekonzentration entlang eines Konvolutes wurde durch Absaugen der perfundierten Lösung in abnehmender Entfernung von der Perfusionsstelle verfolgt. Die pro innere Tubulusoberfläche und Zeit transportierted-Glucosemenge wird mit Abnahme der intraluminalen Glucosekonzentration kleiner. Dieses Verhalten läßt sich durch eine 2-parametrige Gleichung analog der Michealis-Menten-Kinetik beschreiben. Es errechnet sich eine maximale Transportrate,V max, von 6 · 10−10 mol · cm−2 · sec−1 und eine Halbsättigungskonzentration,K m, von 0,6 mmol/l. Die so beschriebene aktive Resorption und die von uns gefundene passive Permeabilität des proximalen Konvolutes fürd-Glucose reichen, nach angestellten Computerberechnungen zu schließen, allein nicht aus, um den Nettoglucosetransport der Gesamtniere unter Freiflußbedingungen quantitativ zu beschreiben.
    Notes: Summary The proximal convoluted tubule of rat kidney was continuously perfused with a steady state solution containing 0.5 to 2.0 mM ofd-glucose. The gradual decrease of intraluminald-glucose concentration was investigated with repeated collections of perfusate from the same tubule whereby the sequence of punctures proceeded towards the site of perfusion. The rate ofd-glucose transport per unit area decreased with decreasing intraluminald-glucose concentration. This relationship could be expressed by a two parameter system corresponding to the Michaelis-Menten equation. It was found that the local maximal transport rateV max equals 6×10−10 mol×cm−2×sec−1 andK m equals 0.6 mM. Our data on active resorption and passive permeability ofd-glucose in the proximal convolution have been subjected to computer analysis. The sum of both components ofd-glucose transport alone as measured under the condition of zero netflux of sodium chloride and water did not match the amount of net glucose transport found for the whole kidney under free-flow-conditions.
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  • 10
    Electronic Resource
    Electronic Resource
    Springer
    Pflügers Archiv 360 (1975), S. 183-187 
    ISSN: 1432-2013
    Keywords: Renal tubule ; Phosphate transport ; pH dependence ; Micropuncture ; Microperfusion
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Early loops of the proximal convoluted tubule of parathyroidectomized rats (PTX-rats) were microperfused with a phosphate (4 mM) containing perfusate. With a perfusion solution of pH around 7.45 as estimated as anion deficit theP i reabsorption was two times greater than with a perfusion solution of pH around 6.85. TheP i reabsorption is reduced in PTX-rats made chronic alkalotic (PTX-cA-rats) but the same pH dependence ofP i reabsorption was found. The data indicate that the divalent phosphate is preferentially reabsorbed.
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