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  • 1
    Electronic Resource
    Electronic Resource
    Springer
    Pflügers Archiv 404 (1985), S. 300-306 
    ISSN: 1432-2013
    Keywords: Epithelial transport ; Contraluminal cell membrane
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract In order to study the specificity for the contraluminal sulfate transport system the inhibitory potency of disulfonates, di-, tricarboxylates and sulfocarboxylates on the35SO 4 2− influx from the interstitium into cortical tubular cells in situ has been determined. The following was found: 1) Methane- and ethane-disulfonate as well as benzene-1,3-disulfonate inhibit contraluminal35SO 4 2− influx (with an (app.K i of 〈6 mmol/l), while benzene-1,2- and 1,4-disulfonate do not. 2) The inhibitory potency of 1,3-benzene disulfonate is slightly augmented by an additional NH2 − or OH-group in position 4. However, OH-groups at position 4 and 5 or 4 and 6 abolish the inhibitory potency. 3) The naphthalene disulfonates tested inhibit only if they have an OH-group in ortho-position to one SO3H group. 4) The stilbene disulfonates H2DIDS and DNDS inhibit the contraluminal35SO 4 2− influx with high (app.K i≈0.8 mmol/l), DADS with lower potency (app.K i≈6 mmol/l). 5) Amongst the tested aliphatic di- and tricarboxylates inhibition was exerted by oxalate (app.K i 1.1 mmol/l) and maleate (app.K i 3.8 mmol/l), but not by malonate, hydroxymalonate and citrate. 6) Out of the tested benzenedicarboxylates only those inhibit which have the COO−-groups directly on the ring in 1,2 and 1,3 position (app.K i 4.0 and 2.7 mmol/l), but not in the 1,4 position. An additional OH-group in position 4 augments the inhibitory potency of 1,3 benzene-dicarboxylates (app.K i 0.8 mmol/l), while an OH group on position 5 abolishes it. 7) The benzene tricarboxylates (BTC) inhibit in the sequence 1,2,3-BTC〉1,3,5-BTC〉1,2,4-BTC (app.K i 0.9, 1.5 and 4.2 mmol/l, respectively). 8) The carboxy-benzene-sulfonates inhibit also in the 1,2 and 1,3 position only (app.K i 6.7 and 5 mmol/l), but not in the 1,4 position. Addition of an −OH-group to the 3-carboxy-1-benzene-sulfonate forming 4-hydroxy-3-carboxy-1-benzene-sulfate augments the inhibitory potency drastically (app.K i 0.32 mmol/l), while a NH2 substitution at the same position leaves it unchanged (app.K i 4.7 mmol/l). If, however, ethylamine instead of NH2 is used as substituent, the inhibitory potency is almost as high as of 4-hydroxy-3-carboxy-1-benzene-sulfonate (app.K i≈0.6 mmol/l). Amongst the dicarboxy-benzene-sulfonates, 3,4-carboxy-benzene-1-sulfonate inhibits (app.K i ca. 2 mmol/l), while 3,5-carboxy-benzene-1-sulfonate does not. The data indicate that a strong interaction of substrate with the sulfate transporter is given, when two charged groups (COO− and/or SO 3 − ) are present in a distance equivalent to the meta-position on the benzene ring and an additional hydrogen bond forming OH- or −NH-group. Hydrogen bond forming groups and charged groups in other positions usually abolish the inhibitory potency.
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  • 2
    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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  • 3
    ISSN: 1432-2013
    Keywords: Oxalate ; Succinate ; Glutarate ; 2-Oxoglutarate ; Citrate ; Sulfate
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract In order to study the specificity for contraluminal para-aminohippurate (PAH) transport, the inhibitory potency of aliphatic dicarboxylates on3H-PAH influx, as well as the inhibitory effect on35SO 4 2− - and3H-succinate influx, from the interstitium into cortical tubular cells in situ has been determined. The following was found: 1. Testing a homologous series of dicarboxylates-ranging from the 2 C oxalate to the 10 C sebacate — PAH transport was inhibited by succinate (app.K i 1.35 mmol/l), and all longer dicarboxylates, with high potency (app.K i 0.05–0.35 mmol/l). Sulfate transport was inhibited only by oxalate (app.K i 1.1 mmol/l), while dicarboxylate transport was inhibited by succinate, glutarate, adipate and pimelate with decreasing potency (app.K i 0.04, 0.24, 0.91, 4.0 mmol/l, respectively). 2. PAH transport was inhibited by succinate and glutarate with high potency (app.K i 1.35 and 0.05 mmol/l), by the correspondent monomethylester to a lesser extent (app.K i 1.7 and 0.74 mmol/l), but not by the dimethylester. On the other hand, the semialdehyde of succinate with aK i-value of 1.2 mmol/l, had the same inhibitory potency as succinate itself, while the dialdehyde of glutarate (app.K i 1.4 mmol/l) was much less potent as glutarate. 3. Introduction of an oxo-, methyl- or sulfhydroxylgroup onto the 2-position of succinate, or of an oxo-group onto the 2-position of glutarate moderately augmented the inhibitory potency against PAH-uptake. However, introduction of a 2-hydroxy group onto succinate or glutarate in thel-position reduced the inhibitory potency more than in thed-position. Introduction of two methyl-, sulfhydryl- or hydroxyl-groups in the 2–3-position of succinate reduced or abolished its inhibitory potency. The introduction of a 2-amino group onto succinate or glutarate abolished its effect on PAH transport. However, N-acetylation or N-benzoylation led to a restitution in inhibitory potency. 4. The trans-isomers fumarate and mesaconate inhibited PAH- and methylsuccinate transport, while the cis-isomers maleate and citraconate did so to a lesser extent or not at all. The effect was reversed with the tricarboxylic aconitates, because cis-aconitate bears a CH2-extended COOH-group in trans-position and trans-aconitate in cis-position. The data indicate that there exist three different anion transport systems at the contraluminal cell side of the proximal renal tubule: 1. a sulfate-oxalate transporter, 2. a sodium-dependent dicarboxylate transporter, and 3. a paraaminohippurate transporter. The PAH transport system accepts dicarboxylates with chain length higher than 7.5 Å (=distance between the terminal oxygen atoms), while the dicarboxylate transport interacts with dicarboxylates with a chain length between 6.5 and 10 Å. Both transport systems prefer the transconfiguration. The effect of side groups on the interaction of dicarboxylates with the PAH-transport system is due mainly to hydrophobicity and electron configuration.
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  • 4
    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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  • 5
    Electronic Resource
    Electronic Resource
    Springer
    Pflügers Archiv 404 (1985), S. 293-299 
    ISSN: 1432-2013
    Keywords: Epithelial transport ; Contraluminal cell membrane
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract In order to study the specificity for the contraluminal sulfate transport system the inhibitory potency of sulfate esters and sulfonate compounds on the35SO 4 2− influx from the interstitium into cortical tubular cells in situ has been determined. The following was found: 1. From 10 sulfate monoesters tested 9 inhibited contraluminal sulfate influx with an app.K i between 0.6 and 6 mmol/l; the two sulfate diesters tested, however, did not. 2. Out of 8 aliphatic sulfonate compounds only three, having a NH- or OH-group in a suitable position, exerted a moderate inhibition (app.K i ca. 2–6 mmol/l). 3. Amongst 14 benzene sulfonates tested only 2 compounds (5-nitrobenzene-sulfonate and 2-hydroxy-5-nitrobenzenesulfonate) inhibited with aK i〈5 mmol/l. 4. Out of 10 naphthalene sulfonates tested 8 inhibited with aK i〈5; the highest inhibition was seen with the NH-containing 8-anilinonaphthalene-1-sulfonate (ANS), but no inhibition with 2 compounds containing an amino group. 5. From the polycyclic sulfonates pyrene-3-sulfonate and anthracene-1-sulfonate inhibited with aK i of approximately 2 mmol/l, while no inhibition was seen with anthracene-2-sulfonate. 6. Out of 4 amino-sulfonates tested benzene-1-amino-sulfonate and a similar benzyl-analog inhibited with aK i of 1 mmol/l and smaller; cyclohexyl-1-amino-sulfonate (cyclamate), however, inhibited only slightly (app.K i of 6 mmol/l). The data indicate that sulfate monoesters are well accepted by the contraluminal sulfate transport system. The affinity of sulfonate compounds to this system depends on neighbouring OH-groups −NH-groups, meta-positioned electronegative groups or a hydrophobic moiety in an appropriate position.
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  • 6
    Electronic Resource
    Electronic Resource
    Springer
    Pflügers Archiv 404 (1985), S. 311-318 
    ISSN: 1432-2013
    Keywords: Epithelial transport ; Contraluminal cell membrane
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract In order to evaluate the specificity for the contraluminal sulfate transport system the inhibitory potency of phenol- and sulfonphthaleins, of sulfamoyl-compounds (diuretics) as well as diphenylamine-2-carboxylates (Cl− channel blockers) on the35SO 4 2− influx from the interstitium into cortical tubular cells in situ has been determined. The following was found: 1) Phenolsulfonphthalein (phenol-red) inhibited with an app.K i-value of 1.7 mmol/l, while analogs which had additional Br-atoms in position 3 and/or 5, i.e. bromphenol-blue, bromcresol-purple and bromcresol-green, inhibited with an apparentK i of 0.1 and 0.5 mmol/l respectively. 2) Phenolphthalein and tetrabromphenolphthalein did not inhibit, while the disulfonate dyes bromsulfalein, fuchsin acid and indigocarmine inhibited with aK i between ≈1 and 3 mmol/l. The highest inhibitory potency in this class of compounds was seen with orange G (app.K i 0.07 mmol/l). The monosulfonate dyes tested, fluoresceinsulfonate and orange I inhibited moderately with an app.K i of ≈5 mmol/l. 3) The 3-sulfamoyl compounds inhibited to a varying degree, when they had a neighbouring −NH-group (furylmethylamino-group), i.e. in position 6 to the COOH or SO3H-group, or when they had a phenoxy-group in position 4. 4) 4-sulfamoylbenzoate and the related compounds probenecid, acetazolamide and hydrochlorothiazide inhibited with an app.K i between 4 and 7 mmol/l. 5) All diphenylamine-2-carboxylate analogs inhibited with an app.K i between 3 and 5 mmol/l, even when the −NH-group was replaced by an =O-group or the benzene ring was replaced by a pyrimidine ring, but not when it was replaced by a thiophen ring. In contrast, 4-phenylaminepyridine-3-sulfonate was ineffective, while diphenylamine-2-amino sulfonate exerted the highest inhibition of this group with an app.K i of 1.4 mmol/l. When, however, the aminosulfonate group was replaced by a methylsulfonamide, the inhibitory potency disappeared. The data can be explained by inhibitory patterns found in previous papers for disulfonates [29], sulfonates with a hydrophobic moiety [28] or neighbouring OH-group [28, 29], carboxylates with a neighbouring −NH- or OH-group in position 2- and an electron-attracting group in position 5 [30].
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  • 7
    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.
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  • 8
    ISSN: 1432-2013
    Keywords: Organic anion transport ; Xenobiotic trans formation ; Structure activity relationship ; ACE inhibitors ; Amanita toxins ; Nephrotoxicity
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract In order to evaluate the specificity of the renal contraluminal PAH transport system for amino acids, oligopeptides and their conjugates, the inhibitory potency of these substances against contraluminal [3H] PAH influx has been determined. For this, inhibition of 3H-PAH flux from the interstitium into cortical tubular cells of the rat kidney in situ has been measured. Apparent Ki values were evaluated by a computer program assuming competitive inhibition. Unconjugated amino acids (glycine, cysteine, alanine, leucine, phenylalanine, tyrosine, aspartate, glutamate, arginine, ornithine and lysine) do not inhibit [3H] PAH influx. The very hydrophobic tryptophan, however, does. N-α-methylation does not change this behaviour. N-α-acetylation does not evoke interaction with the PAH transporter when it occurs with glycine, cysteine (to yield mercapturic acid), arginine, ornithine and lysine. However, it renders alanine, leucine, phenylalanine, tryptophan, L-aspartate moderately, and L-glutamate strongly, inhibitory. The acetylated D-isomers of alanine, leucine and phenylalanine exert a higher inhibitory potency compared with the respective L-isomers. N-α-benzoylation of L-lysine is ineffective. N-α-benzoylation, however, evokes interaction with the PAH transporter, when it occurs with ornithine 〈 arginine 〈 histidine 〈 glycine = leucine 〈 alanine = phenylalanine = aspartate = glutamate. Dipeptides interact with the PAH transporter according to their hydrophobicity (Nozaki scale down to 0.9, Fauchère scale up to 1.0). N-acetylation does not change this behaviour. Hydrophobicity also renders oligopeptides, as angiotensin II, inhibitory against PAH transport. Similarly the anionic angiotensin I converting enzyme inhibitors Captopril, Enalapril and Ramipril inhibit contraluminal PAH influx. The same holds for the Amanita phalloides peptides α- and β-amanitin, phalloin, phallacidin and Tyr5-carboxymethyl antamanide. Conjugation with L-glutathione renders only strongly hydrophobic xenobiotics inhibitory against PAH transport: S-(4-azidophenacyl)-= S-(4-chlorophenacyl)-〈 S-(1,2,2-trichlorovinyl)-〈 S-(1,2,3,4,4-pentachlorobuta-dienyl)- 〈 S-(n-decyl)-. Processing to the L-cysteine conjugate augments the inhibitory potency and additional N-acetylation of the α-amino group augments it even more. Thus, the above mentioned conjugation, which creates hydrophobic molecules with a negative ionic charge, renders it reactive with the PAH transporter. If a remaining positive change at the α-NH 3 + is eliminated by N-acetylation the affinity is further augmented.
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  • 9
    Electronic Resource
    Electronic Resource
    Springer
    Pflügers Archiv 409 (1987), S. 547-554 
    ISSN: 1432-2013
    Keywords: Dicarboxylate transport ; Sulfate transport ; Benzoyl compounds ; Phenoxy compounds ; Valproate
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract In order to study the specificity of the contraluminal para-aminohippurate (PAH) transport system, the inhibitory potency of monocarboxylates on the3H-PAH influx from the interstitium into cortical tubular cells in situ has been determined. The following was found: if a homologous series of fatty acids with increasing chain length is tested, inhibition of contraluminal PAH influx is first seen with valerate (app.K i 1.4 mmol/l), increasing up to nonanoate (app.K i 0.06 mmol/l) and remaining in this range up to duodecanoate, the last compound of this series which is sufficiently water-soluble. Similarly, the inhibitory potency of aromatic monocarboxylates increases with increasing hydrophobicity. If the fatty acids are esterified, their inhibitory potency is lost. If they are transformed to the respective aldehydes their inhibitory potency is preserved at a reduced degree. Introduction of a hydrophobic methyl-, ethyl-, or propyl-group increases the inhibitory potency. A β-, but not an α-oxo-group augments the inhibitory potency of phenylpropionate analogs, an OH group diminishes it, and a NH2 group abolishes it. Among phenyl-fatty acids an increase in affinity is observed from phenyl- 〈 benzoylamine-〈 phenoxy- 〈 benzoyl-acetate and-propionate. All monocarboxylate compounds, so far tested, do not inhibit contraluminal sulfate and Na+/succinate influx. The data indicate that the PAH transporter interacts with monocarboxylates and also with aldehydes which have a hydrophobic moiety. An additional oxo-group facilitates the interaction. Thus, the benzoyl compounds show the highest affinity observed.
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  • 10
    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.
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