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  • 1
    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.
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  • 2
    Electronic Resource
    Electronic Resource
    Springer
    Pflügers Archiv 364 (1976), S. 223-228 
    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).
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  • 3
    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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  • 4
    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.
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  • 5
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    Electronic Resource
    Springer
    Pflügers Archiv 372 (1977), S. 269-274 
    ISSN: 1432-2013
    Keywords: Renal tubule ; Phosphate transport ; Parathyroidectomy ; Parathyroid hormone ; Phosphate diet
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary The standing droplet method was applied in combination with microperfusion of the peritubular blood capillaries to determine the build up of transtubular concentration differences of phosphate (Pi) in proximal convoluted tubules. As revealed in experiments with chronic parathyroidectomized (PTX) rats, the time dependent decrease of the intraluminal Pi concentration, or increase of transtubular Pi concentration difference ( $$\Delta {\text{c}}_{{\text{P}}_i }$$ ), changes along the proximal convolution in a ratio 4:2:1 in the first quarter: second plus third quarter: fourth quarter. In acute (〉2 h) PTX rats $$\Delta {\text{c}}_{{\text{P}}_i }$$ decreased by 31% in the first and by 41% in the fourth quarter of the convolution when parathyroid hormone (PTH; 5 U initially and 12 U/h continuously) was infused. In chronic (〉2 days) PTX rats the correspondent values of 17% and 29% were significantly smaller. When the rats were kept for 7–11 weeks on a low phosphate diet (〈0,15% P in the dry matter) their Pi transport was in the range of that of the PTX rats. PTH infusion, however, diminished the P i reabsorption rate in the fourth quarter of the convolution only, but not that in the early parts of the convolution. On the contrary, rats kept for the same time on a high phosphate diet (2%) showed all along the proximal convolution one by one third of the phosphate transport rate of animals on a low phosphate diet. Acute parathyroidectomy of the high P diet rats led to 51% increase in P i transport. The data show that 1. the phosphate transport decreases as a function of proximal convolution length, 2. PTH exerts a considerable inhibitory effect on P i transport only in acute PTX rats, while the effect in chronic PTX rats is rather small, 3. the P content of the diet inversely correlates with the P i transport. 4. further with low P diet the PTH inhibits P i transport in late, but not in early segments of the proximal convolution.
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  • 6
    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.
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  • 7
    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.
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  • 8
    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.
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