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  • 31
    Electronic Resource
    Electronic Resource
    Active Ca2+ reabsorption in the proximal tubule of the rat kidney (1976)
    Springer
    Pflügers Archiv 364 (1976), S. 223-228 
    Details
    ISSN: 1432-2013
    Keywords: Renal calcium transport ; Renal calcium permeability ; Sodium dependence ; H+ transport ; Ouabain
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Using the stop flow microperfusion technique with simultaneous capillary perfusion the rate of active Ca2+ reabsorption was evaluated by measuring the static head electrochemical potential difference as well as the permeability of the tubular wall for Ca2+ ions. Under control conditions the active Ca2+ transport was calculated to be 3.35×10−13 mol/cm·s. It declined toward zero if the ambient Na+ was replaced by choline or lithium. Parallel experiments in the golden hamster showed that active Ca2+ transport, vanished completely if active Na+ transport was blocked by ouabain (1 mM). These data indicate that the active Ca2+ reabsorption from the proximal tubule depends on the active reabsorption of Na2+ presumably via a Na+−Ca2+ countertransport at the contraluminal cell membrane. The static head electrochemical potential difference of Ca2+ is the same in late and early proximal tubules. It is also not affected by the presence of acetazolamide (10−4 M) by the absence of bicarbonate or glycodiazine buffer or by the absence or presence of phosphate (2 mM).
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00581759
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  • 32
    Electronic Resource
    Electronic Resource
    Coupling between proximal tubular transport processes (1977)
    Springer
    Pflügers Archiv 368 (1977), S. 245-252 
    Details
    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
    URL: http://dx.doi.org/10.1007/BF00585203
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  • 33
    Electronic Resource
    Electronic Resource
    Phosphate transport in the proximal convolution of the rat kidney (1978)
    Springer
    Pflügers Archiv 377 (1978), S. 33-42 
    Details
    ISSN: 1432-2013
    Keywords: Renal tubule ; Phosphate transport ; Extracellular pH ; Intracellular pH ; Acetazolamide
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Inorganic phosphate (Pi) transport was evaluated using the standing droplet method with simultaneous microperfusion of the peritubular capillaries. To evaluate rather small differences in Pi transport and to eliminate the influence of tubular heterogeneity, the technique of crossed paired samples was applied. 1. In chronic PTX rat changing the luminal or both luminal and peritubular pH by varying the HCO 3 − -concentration between 4 and 50 mmol/l at constant 5% CO2 had no influence on Pi transport. 2. If, however, bicarbonate was omitted from the perfusate and 2 mmol/l phosphate (pH 7.4) was the only buffer, Pi transport was decreased from the control. It was, however, further reduced when the perfusates were gased with 5% CO2 i. e. the starting pH was 5.6. 3. When the solutions contained HEPES buffer (25 mmol/l), Pi transport at pH 8 was much larger than at pH 6.0. 4. Raising the CO2 pressure from 35 to 70 mm Hg did not change the Pi transport when both perfusates had a HCO 3 − -concentration of 25 mmol/l. It reduced, however, the Pi transport, when the luminal perfusate had only 4 mmol/l bicarbonate. 5. Lowering the CO2 pressure from 38 to 7.6 mm Hg did hardly change the Pi transport when the luminal perfusate contained 4 mmol/l bicarbonate. It lowered, however, the Pi transport significantly when the luminal perfusate had 25 mmol/l bicarbonate. 6. Acetazolamide, 10−4 M, lowered the Pi transport when the luminal perfusate contained 4 or 25 mmol/l bicarbonate. At 4 mmol/l luminal HCO 3 − , raising thepCO2 to 228 mmol/l depressed Pi transport even more. At 25 mmol/l luminal bicarbonate, raising thepCO2 from 38 to 114 mm Hg reversed the acetazolamide inhibition of the Pi transport almost completely. The data indicate that luminal acidosis and intracellular alkalosis inhibits the transtubular Pi transport. A shift of the intracellular pH to a more alkaline value seems to be responsible for the inhibition of Pi transport by acetazolamide, while omission of buffer from the perfusate inhibits Pi transport by effecting an acidic luminal pH.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00584371
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  • 34
    Electronic Resource
    Electronic Resource
    Reabsorption of monocarboxylic acids in the proximal tubule of the rat kidney (1982)
    Springer
    Pflügers Archiv 395 (1982), S. 212-219 
    Details
    ISSN: 1432-2013
    Keywords: SITS ; Probenecid ; Phloretin ; Acetazolamide ; Lactate ; Renal tubule
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract The transport ofd-lactate across the epithelium of the late proximal convolution was investigated by two methods: 1. by measuring the zero net flux transtubular concentration difference (Δc tt,45s) and the permeability (P) ofd-lactate and calculating from both the transtubular active transport rate (J lac act ). 2. By measuring the 3.5 s efflux ofd-lactate from the tubular lumen, while blood was flowing through the capillaries. The 3.5 s efflux comprises two components, one going through the brush border (J lac bb ) and one going the paracellular pathway (J lac paracell =P lac·c lac lumen). Both,J lac act andJ lac bb ofd-lactate gave the sameK m 1.9 and 1.7 mmol/l and the same maximal transport rate 3.2 and 2.9 pmol cm−1 s−1. TheK i ofl-lactate tested againstJ lac act andJ lac bb ofd-lactate was also the same: 1.1 and 1.0 mmol/l. These data indicate that under our experimental conditions only the flux through the brush border seems to be rate limiting and thatd-lactate uses the same transport system asl-lactate. When Na+ was omitted from the perfusatesJ lac act disappeared completely, whileJ lac bb was reduced by 64%. These data reflect the Na+ dependence of thed-lactate transport through the brush border. Variation of intra-and extracellular pH by raisingpCO2, omitting HCO 3 − from the perfusates or adding acetazolamide had no effect on the transport ofd-lactate when α-ketoglutarate was used as fuel. However, when acetate was used as fuel, intracellular acidosis brought the reducedJ lac act back to the values obtained with α-ketoglutarate as fuel. It is suggested that this is an effect on a contraluminal transport step. Probenecid (5 mmol/l) and phloretin (0.25 mmol/l) inhibitedJ lac act significantly.J lac bb , however, was only inhibited by probenecid when acetate was used as fuel. These data indicate that both compounds act on thed-lactate exit at the contraluminal cell side, but that probenecid acts in addition at the luminal cell side. SITS (1 mmol/l) augmentedJ lac bb when acetate was used as fuel and is similar to the effect of lowering intracellular pH as described above. The SH reagents mersalyl (1.0 mmol/l) and maleolylglycine (1 mmol/l) did not influenceJ lac bb .
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00584812
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  • 35
    Electronic Resource
    Electronic Resource
    Renal phosphate transport: Inhomogeneity of local proximal transport rates and sodium dependence (1975)
    Springer
    Pflügers Archiv 356 (1975), S. 287-297 
    Details
    ISSN: 1432-2013
    Keywords: Renal Tubule ; Phosphate Transport ; Sodium Dependence ; Micropuncture ; Microperfusion
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary The standing droplet method has been used in combination with the peritubular perfusion of blood capillaries to determine the build up of transtubular concentration differences of phosphate (P i ) in the renal proximal convoluted tubule of parathyroidectomized rats. Electron probe analysis was used to estimate P i . At zero time both the intraluminal and the contraluminal P i concentration was 2 mM. The time dependent decrease of the intraluminal P i concentration was approximately 4 times faster in the early than in the late proximal convoluted tubule. After 45 sec an intraluminal steady state concentration of 0.20 mM P i was achieved in the early part. In the late part the intraluminal P i concentration approached a steady state value of 0.54 mM at 120 sec. When sodium free solutions were used the intraluminal P i concentration increased to 2.22 mM in the earlier and to 2.76 mM in the late part. The data indicate that in the proximal convoluted tubule 1. The rate of phosphate reabsorption is greater in the early part than in the later part, and 2. phosphate reabsorption might occur as co-transport with Na+ ions.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00580003
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  • 36
    Electronic Resource
    Electronic Resource
    Renal proximal tubular buffer-(glycodiazine) transport (1975)
    Springer
    Pflügers Archiv 357 (1975), S. 149-163 
    Details
    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
    URL: http://dx.doi.org/10.1007/BF00585971
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  • 37
    Electronic Resource
    Electronic Resource
    Effect of SH-, NH2- and COOH-site group reagents on the transport processes in the proximal convolution of the rat kidney (1973)
    Springer
    Pflügers Archiv 344 (1973), S. 51-68 
    Details
    ISSN: 1432-2013
    Keywords: Proximal Kidney Tubule ; Mercurials ; SH Reagents ; Site Group Reagents ; Transtubular Transport
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary The effects of site group reagents were tested on the following transport processes of the proximal convolution. Isotonic Na+ absorption, evaluated by the shrinking droplet procedure, histidine and glucose transport, evaluated by measuring the respective transtubular concentration difference at zero substance and water net flux. The test substances were applied either by continuous microperfusion of the peritubular capillaries or by luminal perfusion prior to the transport tests or by addition to the luminal test solution. The SH reagents (0.2 mM) N-ethylmaleimide,p-chloromercuribenzoate (pCMB) 3,6-bis-(acetatomercurimethyl)dioxane and Mersalyl (Salyrgane) caused 50% inhibition of the isotonic Na+ absorption in approximately 1.5 min when applied to the capillary perfusate. The same effect was reached in 2–3 min by 0.2 mMp-chloromercuriphenylsulfonate, benzamido-4-iodo-acetylstilbene-2,5-disulfonate and 2,2′-dihydroperoxy-2,2′-dibutylperoxide. However, the large molecular SH reagentspCMB-dextran T10 and benzoxanthene-3,4-dicarboxylic-N-iodoacetyloligoprolyl-2-aminoethylimid, did not inhibit the isotonic Na+ absorption. If an inhibitory effect was observed on the Na+ transport its onset was faster, when the substance was applied from the blood site than when it was given from the tubular lumen. Because SH reagents inhibit the isotonic Na transport faster when applied from the blood side, and because SH reagents with MW up to 690 are inhibitory whereas larger ones with MW over 1700 are not, it seems that they exert their inhibitory action on SH groups located a) predominantly on the blood side and b) deep within the membrane and not at the surface. Histidine- and glucose transport was inhibited only when the sodium transport was inhibited considerably. The oxygen consumption of teased kidney slices is not inhibited by 0.2 mMpCMB or Mersalyl within 10 min, but it is inhibited considerably by 1 mM of these substances in the same period of incubation time. The COOH reagents N,N′-carbonyl-diimidazole and N-ethyl-N′-(3-dimethyl-aminopropyl)carbodiimid (10 mM) and the NH2 reagents 4-acetamido-4′-isothiocyanatostilbene-2,2′-disulfonic acid, 2 Na+ (SITS) (1 mM) as well as danslychloride (applied from the lumen at 5 mM in paraffin oil) did not inhibit the isotonic Na+ absorption.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00587441
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  • 38
    Electronic Resource
    Electronic Resource
    Phosphate transport in the proximal convolution of the rat kidney (1978)
    Springer
    Pflügers Archiv 375 (1978), S. 97-103 
    Details
    ISSN: 1432-2013
    Keywords: Renal tubule ; Phosphate transport ; Paracellular shunt ; Calcium ; Ca2+ ionophore A 23187
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Proximal inorganic phosphate (P i ) transport was evaluated using the standing droplet method with simultaneous microperfusion of the peritubular blood capillaries. In chronic parathyroidectomized (PTX) rats addition of 3 μM of the Ca2+ ionophore A 23187 to the luminal perfusate had no effect on the P i transport, although the isotonic fluid reabsorption was reduced by 20%. When the Ca2+ concentration in the perfusates was raised from 1.5 mM to 3.0 mM the reabsorption did not change significantly. But when Ca2+ was omitted from the perfusates the P i reabsorption dropped by 19%, and when 2 mM EDTA were added to the perfusates P i transport decreased by 35%. The influx of P i from the interstitial space and from the cell into the phosphate-free luminal perfusate did not change, when the perfusates were Ca2+-free, but it increased by 23% in the presence of 2 mM EDTA. The data indicate that 1. a rise in intracellular Ca2+ above normal is not a factor which modifies “basal” P i transport i.e. when P i transport is independent of the action of parathyroid hormone. 2. A reduction of extracellular Ca2+ concentration from normal toward zero reduces P i transport without changing the paracellular leak permeability for P i . 3. With EDTA the paracellular leak permeability for P i is increased, thus causing an even greater reduction in net P i transport than with Ca2+-free solutions alone.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00584153
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  • 39
    Electronic Resource
    Electronic Resource
    Active sulfate reabsorption in the proximal convolution of the rat kidney: Specificity, Na+ and HCO 3 − dependence (1980)
    Springer
    Pflügers Archiv 383 (1980), S. 159-163 
    Details
    ISSN: 1432-2013
    Keywords: Renal tubule ; Sulfate transport ; Na+ coupled transport ; Thiosulfate ; Molybdate
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Using the standing droplet technique in the proximal convolution and simultaneous microperfusion of the peritubular capillaries, the decrease in luminal sulfate concentration with time and the zero net flux transtubular concentration difference of sulfate ( $$\Delta c_{{\text{SO}}_{\text{4}}^{{\text{2 - }}} } $$ ) at 45 s was determined — the latter being taken as a measure of the rate of active sulfate reabsorption. Starting with 0.5 mmol/l sulfate in both perfusates the $$\Delta c_{{\text{SO}}_{\text{4}}^{{\text{2 - }}} } $$ value of 0.35 mmol/l was approached exponentially with a half value time of 4.3 s. The $$\Delta c_{{\text{SO}}_{\text{4}}^{{\text{2 - }}} } $$ values in the early proximal and late proximal convolution did not deviate from each other. If the Na+ concentration in the perfusates was reduced, the $$\Delta c_{{\text{SO}}_{\text{4}}^{{\text{2 - }}} } $$ approached zero and extrapolated to a slightly negative value (c i〉c o). When 1 mmol/l ouabain was added to the perfusates $$\Delta c_{{\text{SO}}_{\text{4}}^{{\text{2 - }}} } $$ decreased by 66% (the latter experiments were performed in the golden hamster which is more sensitive to ouabain than the rat). 1 mmol/l thiosulfate diminished $$\Delta c_{{\text{SO}}_{\text{4}}^{{\text{2 - }}} } $$ by 68% and 1 mmol/l molybdate by 24%. Omitting or replacing bicarbonate by HEPES or glycodiazine reduced the sulfate reabsorption significantly, while acetazolamide (0.1 mmol/l) and increasing the CO2-pressure from 4.66 to 14.0 kPa (i.e. 5–15% CO2) had no effect. SITS 1 mmol/l had no effect on sulfate reabsorption. The data indicate that the sulfate reabsorption is driven by a Na+ gradient and inhibited by thiosulfate and molybdate, i.e. molecules which have a similar tetrahedral molecule structure. The sulfate reabsorption depends in an undefined manner on the presence of bicarbonate ions.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00581877
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  • 40
    Electronic Resource
    Electronic Resource
    A cytotoxin of pseudomonas aeruginosa acts directly on the cortical thick ascending limb of henle's loop of rabbit kidney (1982)
    Springer
    Pflügers Archiv 394 (1982), S. 271-273 
    Details
    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
    URL: http://dx.doi.org/10.1007/BF00589103
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