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  • 1
    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.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1432-2013
    Keywords: 2-Oxoglutarate ; Lactate ; Pyruvate ; Nitrate
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract In order to study the characteristics of contraluminal para-aminohippurate transport into proximal tubular cells the stopped flow capillary perfusion method was applied. The disappearance of3H-paraaminohippurate from the capillary perfusate at different concentrations and contact times was measured and saturation type behaviour was found with aK m of 0.08±0.01 (SE) mmol/l,J max of 1.1±0.1 pmol·s−1·cm−1 andr, the final extracellular/intracellular distribution ratio of 0.93±0.03. Omission of Na+ from the capillary test perfusate caused a small reduction of contraluminal PAH uptake at small transport rates (0.1 mmol/l PAH in the test perfusate) but not at high transport rates (1.0 mmol/l PAH in the test perfusate). Change of K+ between 0 and 40 mmol/l and pH between 6.0 and 8.0 did not influence contraluminal PAH uptake. Isotonic replacement of chloride by gluconate, nitrate, sulfate, phosphate, methanesulfonate or increase in bicarbonate to 50 mmol/l did not influence PAH uptake at small transport rates. But isotonic sulfate and phosphate, as well as 50 mmol/l HCO 3 − and 25 mmol/l Hepes in isotonic solutions reduced PAH uptake at high transport rates. Addition of 5 mmol/l Ca2+, Mg2+, Mn2+, Ba2+, Cd2+ to isotonic Na+-gluconate solution did not influence PAH uptake except for Mg2+ and Mn2+ which inhibited uptake at small transport rates only. Preperfusion of the peritubular capillaries with rat serum, Na+ gluconate (Ca2+-+Mg2+-free), Na+ gluconate (Ca2+-+Mg2+-free) plus 10 mmol/l lactate or pyruvate or 0.1 mmol/l 2-oxoglutarate did not influence PAH uptake at small PAH transport rates, but inhibited at high transport rates. Preperfusion of the capillaries for 10 s with Na+-, Ca2+- and Mg2+-free solutions reduced PAH uptake in the presence of Na+ at both transport rates. A second 10 s preperfusion — after the first 10 s Na+-, Ca2+-, Mg2+-free preperfusion — with serum or solutions which contained Na+ and Ca2+ or Mg2+ restored the PAH fluxes to control values. The data are compatible with the hypothesis that contraluminal PAH uptake occurs by a saturable transport mechanism in exchange for other intracellular anions rather than in cotransport with Na+ ions. It was, however, not possible to identify the type of counteranions involved. The large effect of cation replacement on para-aminohippurate transport, which was reported in many previous studies with kidney slices, is not a direct effect on the para-aminohippurate transporter, but is rather caused indirectly via cell metabolism and/or changed ion gradients.
    Type of Medium: Electronic Resource
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  • 3
    Electronic Resource
    Electronic Resource
    Springer
    Pflügers Archiv 410 (1987), S. 501-504 
    ISSN: 1432-2013
    Keywords: Na+-dependence ; Cl−-dependence ; Sulphate dependence ; DIDS ; Carbonic anhydrase inhibitors ; Nitrophenylglyoxal
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract In order to measure the contraluminal bicarbonate flux in situ we applied the stopped flow capillary microperfusion technique and measured the influx of14C-bicarbonate buffer into cortical tubular cells at pH 8. It was found that the influx in percent of the starting concentration is larger at 20 mmol/l bicarbonate than at 1 mmol/l, indicating a sigmoidal type influx curve. At 20 mmol/l bicarbonate the influx was inhibited by 44%, when Na+ was replaced by choline. Replacement of gluconate by chloride or sulfate did not change H14CO 3 − influx. At this bicarbonate concentration, influx is inhibited by 10 mmol/l 4,4′-diisothiocyanato-2,2′-stilbenedisulfonate (DIDS) (22%), 5 mmol/l of the carbonic anhydrase blocker ethoxyzolamide (40%) as well as by 5 mmol/l of the arginine reagent 4-nitrophenylglyoxal (31%). At 1 mmol/l bicarbonate starting concentration, bicarbonate influx was inhibited when chloride in the perfusate was present or when sulphate was added. Replacement of sodium by choline did not change bicarbonate influx. Addition of DIDS and 8-anilino-naphthalene-1-sulfonate (5 mmol/l each) inhibited 1 mmol/l bicarbonate influx 39 and 49%, respectively. The para-aminohippurate transport blocker dipropylsulfamoyl-benzoate (probenecid), the chloride channel blocker 5-nitro-2′-(3-phenylpropylamino)-benzoate (NPPB), the SH group blocker 2-(3-hydroxymercuri-2-methoxypropyl)-carbamoyl-phenoxyacetate (mersalyl), and formate did not inhibit bicarbonate influx, at 20 and at 1 mmol/l H14CO 3 − starting concentration. The data are compatible with the assumption of 1. a contraluminal (HCO 3 − )3/Na+ cotransporter inhibitable by DIDS, carbonic anhydrase inhibitors and 4-nitrophenylglyoxal, 2. a HCO 3 − /anion exchange system, which accepts sulfate and chloride and is inhibitable by the anion exchange blockers DIDS and 8-anilino-naphthalene-1-sulfonate, and 3. a HCO 3 − influx component which could not be influenced by Na+, Cl−, nor by the inhibitors applied.
    Type of Medium: Electronic Resource
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