Effects of Interactions Between Drugs on the Renal Excretion of Trientine in Rats—Acetazolamide and Furosemide Increase Trientine Excretion

Purpose. To elucidate the effects of drug interactions on the urinary excretion of trientine in rats. Methods. Trientine and various other drugs were intravenously administered to rats and the urinary excretion of trientine was investigated. To clarify the mechanisms of drug-drug interactions, we also investigated the effects of various drugs on spermine uptake by rat renal brush-border membrane vesicles. Results. Cimetidine, a substrate of the H⁺/organic cation antiporter, and aminoglycoside antibiotics did not affect trientine excretion, while acetazolamide and furosemide, which increase the concentration of sodium ions in renal proximal tubules, increased the excretion of trientine. However, trichlormethiazide, which acts in renal distal tubules, did not affect trientine excretion. Acetazolamide and furosemide did not directly affect the Na⁺/spermine transporter because these diuretics had no effect on the uptake of spermine into the rat renal brush-border membrane vesicles. Conclusions. There is no interaction between trientine and the substrate of the H⁺/organic cation antiporter or aminoglycoside antibiotics. However, drugs that change the concentration of sodium ions in renal proximal tubules, such as diuretics, can increase the trientine excretion since the increase in the luminal concentration of sodium ion accelerates the Na⁺/spermine antiporter.     

The presence of an Na+/spermine antiporter in the rat renal brush-border membrane.

This study was aimed at determining the driving force for spermine transport in rat renal proximal tubular brush-border membrane. The uptake of spermine and trientine, a spermine-like drug used for treating Wilson's disease, into rat renal brush-border membrane vesicles was significantly stimulated by an outwardly directed Na+ gradient. The Na+-dependent uptake was temperature dependent and saturable. A kinetic analysis of the initial uptake of spermine with an Na+ gradient gave a Km value of 1.44 microM and a Vmax value of 6.31 pmol (mg protein)(-1)/30s. The Na+ dependent uptake of [3H]spermine was inhibited by spermine, trientine and tetraethylene-pentamine. Substrates of the H+/organic cation transporter (cimetidine and tetraethyl-ammonium), physiological polyamines (putrescine and spermidine) with 2 or 3 amino groups and aminoglycosides (amikacin and tobramicin) with 4 or 5 cationic amines did not affect the uptake of spermine in the presence of an outwardly directed Na+ gradient. These results suggest that the renal tubular secretion of spermine is mediated by an Na+/spermine antiport system which is specific for a straight-chain polyamine compound with more than 4 amino groups.     

Uptake Mechanism of Trientine by Rat Intestinal Brush-border Membrane Vesicles

The uptake characteristics of trientine by rat intestinal brush-border membrane vesicles were studied. The uptake characteristics of trientine were similar to those of the physiological polyamines with respect to the excessive accumulation in vesicles, the pH dependency, the temperature dependency and the ineffectiveness of K⁺ diffusion potential (inside negative). The initial uptake of trientine was saturable with a K_m value of 1.13 mM, which was larger than that of spermine and spermidine. Furthermore, the uptake rate of trientine was dose-dependently inhibited by spermine and spermidine. Spermine competitively inhibited the uptake of trientine with a K_i value of 18.6 μM, and it was close to the K_m value for spermine (30.4 μM). These data suggested that the uptake of trientine was similar to that of spermine and spermidine in rat small intestinal brush-border membrane vesicles, and these polyamines seem to inhibit the absorption of trientine from the gastrointestinal tract.     

The transport mechanism of polycationic compounds across intestinal and renal cell membrane

This article reviewed the transport mechanism of polycationic compounds across rat intestinal and renal cell membranes. The inside-negative diffusion potential stimulated the initial uptake of dicationic compounds into intestinal brush-border membrane vesicles, and a good correlation was observed between lipophilicity and the amount of diffusion potential-dependent transport of the dications. On the other hand, tri- and tetracationic compounds were not affected by the diffusion potential because of their much lower lipophilicity. The membrane surface potential affected to the transport of polycationic compounds, similar to monocationic compounds. Therefore it appears that the membrane surface potential plays a common role in the transport of mono- and polycationic compounds across cell membranes. On the intestinal basolateral membrane, it was found that there was a Na+/putrescine symporter. This recognized dicationic compounds and transported them from the blood into intestinal cells. This transporter did not recognize spermine and spermidine. Furthermore, we found a novel transport system, a Na+/spermine antiporter, on the rat renal brush-border membrane. This transporter recognized aliphatic polycation, which has more than four amino groups, and actively secreted spermine and trientine into the renal proximal tubules in vitro and in vivo. However, this transporter did not recognize trientine-copper complex. These results are useful for the prediction of the intestinal absorption and renal excretion of polyamine derivatives.     

Phenolsulfonphthalein transport by potential-sensitive urate transport system

The purpose of this study was to elucidate the transporter-mediated secretion systems for phenolsulfonphthalein in brush-border membranes. In human and rat renal brush-border membranes, a potential-sensitive transport system has been shown to be involved in the efflux of organic anions. The uptake of phenolsulfonphthalein into rat renal brush-border membrane vesicles was stimulated by an inside-positive membrane potential. This potential-sensitive uptake of phenolsulfonphthalein was inhibited by probenecid, pyrazinoate and urate. p-Aminohippurate had no effect on the potential-sensitive uptake of phenolsulfonphthalein. Moreover, urate competitively inhibited the uptake of phenolsulfonphthalein. On the other hand, the uptake of phenolsulfonphthalein was slightly increased in the presence of an outward Cl- gradient. These results suggest that phenolsulfonphthalein has high affinity for the potential-sensitive urate transport system but has low affinity for an anion exchanger.     

Renal handling of biphosphonate alendronate in rats.

Alendronate is a bisphosphonate that is secreted via a saturable pathway in rat kidney. This study is designed to discover if the rate-determining step in its net renal secretion is uptake into the renal tubule. The tissue uptake clearance of alendronate by the kidney, estimated from an integration plot analysis and normalized with respect to plasma protein binding, was 4.2 times higher at a tracer dose than that of inulin, indicating uptake of alendronate by the renal tubules. The uptake clearance is comparable with the net secretion clearance obtained from an infusion study, indicating that the rate-determining step in the net secretion is uptake under the tracer conditions. When the dose was increased, however, there was no reduction in uptake clearance while the net secretion clearance fell to almost zero. The urinary excretion clearance defined with respect to the steady state concentration in the kidney also fell to almost zero. This result suggests that saturation of the net secretion of alendronate is caused by saturation of membrane transport through the brush-border membrane. Thus, it would seem that there is a transport mechanism for alendronate on the brush-border membrane of kidney epithelial cells.     

Transport Mechanism of Ceftibuten,a Dianionic Cephem,in Rat Renal Brush-Border Membrane

The uptake mechanism of ceftibuten by rat renal brush-border membrane vesicles was investigated. Uptake was found to be independent of a Na⁺ gradient and partially dependent on an inwardly directed H ⁺-gradient. Competition experiments between ceftibuten and several compounds demonstrated that the peptide-like structural features of inhibitors are more essential than their charge properties for inhibiting uptake. Anionic compound, such as p-amino-hippuric acid, also inhibited ceftibuten uptake by renal brush-border membrane vesicles in the presence of an H ⁺-gradient. We conclude that ceftibuten, in spite of its anionic structure, is transported via the dipeptide transport systems, rather than the organic anion transport system.     

The Transport Mechanism of an Organic Cation,Disopyramide, by Brush-border Membranes. Comparison Between Renal Cortex and Small Intestine of the Rat

Abstract— The characteristics of disopyramide uptake in brush-border membrane vesicles isolated from rat renal cortex and small intestine were investigated. Transport of disopyramide into an osmotically reactive intravesicular space was observed with notable binding to the membrane surface. An outwardly directed H⁺ gradient stimulated disopyramide uptake, resulting in a transient uphill transport in both brush-border membranes. As for the renal brush-border membrane, the H⁺ gradient itself appeared to be the driving force for this stimulation of uptake. These findings suggest that disopyramide-H⁺ antiport is the mechanism of disopyramide action in renal cell membrane. The initial uptake was saturable (K_m and V_max of 680 μm and 1·25 nmol (mg protein)^?1/30 s, respectively). The stimulation of disopyramide uptake by an outward H⁺ gradient in rat intestinal brush-border membrane was due to an interior negative H⁺-diffusion potential. A K⁺-diffusion potential (interior negative) enhanced disopyramide uptake. These results suggest that there are different mechanisms of disopyramide uptake for renal and intestinal brush-border membrane vesicles.     

The influence of trimethoprim,sulfamethoxazole, and creatinine on renal organic anion and cation transport in rat kidney tissue

The use of combined trimethoprim and sulfamethoxazole has been reported to increase serum creatinine concentrations in patients by inhibiting creatinine excretion. In confirmation, we found that trimethoprim given over 24 hr to rats augmented serum creatinine concentrations and decreased creatinine clearance relative to urea clearance. Previously, decreased urinary excretion has been hypothesized to occur secondarily to inhibited renal secretory transport of creatinine via the organic cation pathway, although there was no direct evidence to indicate that trimethoprim inhibits organic cation secretion. Such a mechanism is possible because other studies have shown that creatinine is secreted through both the organic anion and cation pathways. We followed organic anion ([³H]PAH) and organic cation ([¹⁴C]TEA) accumulation in incubating rat renal fragments in order to determine whether inhibition of organic cation accumulation by trimethoprim actually does occur. The present investigation shows that trimethoprim interferes with organic cation uptake in vitro but does not influence organic anion uptake. Sulfamethoxazole affected neither organic cation nor anion accumulation significantly. Trimethoprim decreased influx of [¹⁴C]TEA, not its efflux. Other metabolic processes such as QO₂, ammoniagenesis, and gluconeogenesis were not altered greatly by trimethoprim. Creatinine itself decreased organic anion and cation transport through competitive inhibition. Added to the findings that trimethoprim elevates serum creatinine in man without changing glomerular filtration rate, our results strengthen the hypothesis that trimethoprim can decrease serum creatinine excretion through the inhibition of creatinine secretion via the organic cation transport route.     

Ionic-diffusion Potential-dependent Transport of a New Quinolone,Sparfloxacin, Across Rat Intestinal Brush-border Membrane

The mechanism of uptake of sparfloxacin, a new quinolone, by intestinal brush-border membrane vesicles was investigated to clarify whether there is a common transport process for new quinolones mediated by the diffusion potential across the intestinal membrane bilayer. Sparfloxacin was taken up pH-dependently by rat intestinal brush-border membrane vesicles, behaviour analogous to that of organic cations including enoxacin and ciprofloxacin. Transient overshooting uptake of this quinolone was observed in the presence of an outward H+ gradient. Momentary dissipation of the H+ gradient by addition of carbonyl cyanide p-(trifluoromethoxy)phenylhydrazone did not affect the uptake of sparfloxacin, and a marked but incomplete reduction in the H+-sensitive overshooting uptake of sparfloxacin was apparent in the voltage-clamped brush-border membrane vesicles. Furthermore, a valinomycin-induced K+-diffusion potential (interior negative) and an inward Cl^—-diffusion potential stimulated the initial uptake of Sparfloxacin at pH 5.5. Sparfloxacin uptake was inhibited by tetracaine and imipramine. The inhibitory effect of these cations correlated well with changes in membrane surface charges induced by the presence of tetracaine or imipramine. These results indicate that sparfloxacin transport across the brush-border membrane depends upon the inside-negative ionic diffusion potential, that the H+- or K+-diffusion-potential-dependent uptake of sparfloxacin by intestinal brush-border membrane vesicles is affected by the membrane surface potential and that inhibition of sparfloxacin uptake originates from changes in the membrane surface potential caused by the organic cations.     

Effect of lamivudine on uptake of organic cations by rat renal brush-border and basolateral membrane vesicles

The effect of lamivudine on uptake of a representative organic cation, tetraethylammonium (TEA), by rat renal brush-border membrane vesicles (BBMV) and basolateral membrane vesicles (BLMV) has been investigated. The pH-driven uptake of TEA by BBMV (pHin = 6.0, pHout = 7.5) was inhibited by lamivudine. The IC50 value (concentration resulting in 50% inhibition) for the concentration-dependent effect of lamivudine on TEA uptake by BBMV after 30 s was 2668 microM whereas IC50 values for cimetidine and trimethoprim were < 2.5 microM and < 25 microM, respectively. The early uptake of TEA by BLMV was also reduced significantly by lamivudine. The IC50 value for the concentration-dependent effect of lamivudine on uptake of TEA by BLMV at 30 s was > 25 mM, whereas the IC50 values for cimetidine and trimethoprim were 2116 microM and 445 microM, respectively. These findings suggest that compared with other cationic drugs, such as trimethoprim and cimetidine, lamivudine is a weak inhibitor of organic cation transport into the tubules by the brush-border and basolateral membranes of renal epithelial cells. It is unlikely lamivudine will have any significant effect on the excretion of co-administered cationic drugs by the renal tubules.     

Transport characteristics of cephalosporin antibiotics across intestinal brush-border membrane in man, rat and rabbit.

The uptake of orally active cephalosporins, ceftibuten and cephradine, by intestinal brush-border membrane vesicles isolated from man, rat and rabbit was studied. In the presence of an inward H+ gradient, ceftibuten but not cephradine was taken up into intestinal brush-border membrane vesicles of man and rat against the concentration gradient (overshoot phenomenon). In rabbit jejunal brush-border membrane vesicles, the uptake of both cephalosporins in the presence of an inward H+ gradient exhibited the overshoot phenomenon. In human and rat vesicles, the initial uptake of ceftibuten was strongly inhibited by compound V, an analogue of ceftibuten, but the uptake of cephradine was not affected by any of the cephalosporins tested, whereas in the rabbit brush-border membrane vesicles, initial uptake of both ceftibuten and cephradine were markedly inhibited by all cephalosporins and dipeptides used. These results suggest that the transport characteristics of human and rat intestinal brush-border membrane for cephalosporins are comparable, and that rabbit is an inadequate animal for investigating the transport characteristics of beta-lactam antibiotics.     

Modulation of Organic Cation Transport and Lipid Fluidity by Benzyl Alcohol in Rat Renal Brush-border Membranes

Purpose. Organic cations are actively transported in renal brush-border membranes (BBM) by the H⁺/organic cation antiport system. In the present study, we investigated the relationship between membrane fluidity and organic cation transport in the BBM. Methods. The effects of benzyl alcohol, a membrane fluidizing agent, on the organic cation tetraethylammonium (TEA) uptake were studied using renal BBM vesicles isolated from rat kidney. BBM fluidity was assessed by fluorescence polarization technique. Results. H⁺ gradient-dependent uptake of TEA in BBM vesicles was inhibited by benzyl alcohol in a dose-dependent manner, with an apparent half inhibitory concentration of 18mM. The decrease in fluorescence anisotropy of l,6-diphenyl-l,3,5-hexatriene in BBM, which represents the increase in membrane fluidity, was correlated with the decrease in TEA transport activity. The dissipation rate of H⁺ gradient, a driving force for organic cation transport in BBM, was increased by benzyl alcohol. In addition, H⁺ gradient-independent TEA-TEA exchange was also inhibited by benzyl alcohol. These findings indicate that benzyl alcohol inhibits the uptake of TEA by affecting the intrinsic activity of the organic cation transporter and the H⁺ gradient dissipation rate. Conclusions. The membrane fluidity should be an important determinant for organic cation transport in renal BBM.     

D‐Malate decreases renal content of α‐ketoglutarate,a driving force of organic anion transporters OAT1 and OAT3, resulting in inhibited tubular secretion of phenolsulfonphthalein,in rats

d ‐Malate inhibits a Krebs cycle enzyme and the tubular transport of α‐ketoglutarate, an intermediate of the Krebs cycle and the driving force for rat organic anion transporter 1 (rOAT1) and rOAT3 in the kidney. This study examined the effects of d ‐malate on the rat organic anion transport system. The uptake of 6‐carboxyfluorescein by HEK293 cells expressing rOAT1 or rOAT3 was not affected by d ‐malate and l ‐malate. Up to 60 min after the intravenous injection of phenolsulfonphthalein (PSP), a typical substrate of the renal organic anion transporters, as a bolus to rats, 47.1% of the dose was recovered in the urine, and its renal clearance was estimated to be 8.60 ml/min/kg. d ‐Malate but not l ‐malate interfered with its renal excretion, resulting in the delayed elimination of PSP from plasma. No effect of d ‐malate was recognized on creatinine clearance or the expression level of rOAT3 in the kidney cortex. d ‐Malate increased the plasma concentration of α‐ketoglutarate. In addition, the compound greatly stimulated the renal excretion of α‐ketoglutarate, implying that d ‐malate inhibited its reabsorption. The content of α‐ketoglutarate was significantly decreased in the kidney cortex of rats administered d ‐malate. Collectively, this study shows that d ‐malate abrogates the tubular secretion of PSP, and the reduction of the renal content of α‐ketoglutarate was proposed to be one of the mechanisms. A relationship between the reabsorption of α‐ketoglutarate and the basolateral uptake of organic anion in the kidney is suggested.     

H+ gradient-dependent transport of aminocephalosporins in rat intestinal brush-border membrane vesicles. Role of dipeptide transport system

The transport of cephalosporin antibiotics in brush-border membrane vesicles isolated from rat small intestine has been studied by a rapid filtration technique, demonstrating a carrier-mediated transport system for aminocephalosporins such as cephradine. In agreement with the transport mechanisms of dipeptides, the uptake of cephradine by brush-border membrane vesicles was Na+-independent and was stimulated in the presence of an inward H+ gradient ([H+]o greater than [H+]i). Cephradine uptake in the presence of an inward H+ gradient was a saturable process with an apparent Km of 9.4 mM, and was markedly inhibited by dipeptides but not inhibited by amino acids. The present data suggest that aminocephalosporins can be transported by a common carrier-mediated system with dipeptides in the intestinal brush-border membranes and this process may be driven by an H+ gradient.     

Effect of Renal Ischaemia on Organic Compound Transport in Rabbit Kidney Proximal Tubule

Abstract: This study was carried out to determine the effect of renal ischaemia on transport systems for organic compounds in the rabbit kidney proximal tubule. Ischaemia for 30 or 60 min. induced glucosuria and phosphaturia, which was accompanied by polyuria and natriuresis. The Na⁺-dependent uptake of glucose, succinate and L-glutamate by brush-border membrane vesicles was not altered by 30 or 60 min. of ischaemia, while the H⁺/tetraethylammonium antiport was significantly inhibited after 30 min. of ischaemia. When the duration of ischaemia was extended to 120 min. the uptake of glucose and succinate by brush-border membrane vesicles was also significantly attenuated, but the L-glutamate uptake was not altered. The uptake of glucose, succinate and L-glutamate by basolateral membrane vesicles was not impaired even with 120 min. of ischaemia, suggesting that transport systems for organic compounds in the brush-border membrane are more sensitive to ischaemia than those in the basolateral membrane. Ouabain-sensitive oxygen consumption in renal cortical slices was not depressed by 60 min. of ischaemia. When kidneys were reperfused for 60 min. following 60 min. of ischaemia, the Na⁺-glucose and Na⁺-succinate cotransport and the H⁺/tetraethylammonium antiport were not different from the control, but the recovery of alkaline phosphatase was significantly reduced. When kidneys were subjected to ischaemia for 60 min., a loss of brush-border microvilli and plasma membrane was observed after 5 or 60 min. of reflow in the proximal convoluted tubule. After 3 hr of reflow, focal necrosis appeared although the microvilli were partially regenerated. These results indicate that renal ischaemia induces selective impairments of transport systems in the rabbit renal proximal tubule, and that the glucosuria and phosphaturia induced by transient ischaemia are results, at least in part, from a diminution of brush-border surface area rather than a direct defect in the function of the membrane transport systems.     

Renal excretion mechanism of NS-49, a phenethylamine class alpha 1A-adrenoceptor agonist.

Renal handling of NS-49, which is an organic cation and a chiral compound, was investigated in rats, rabbits and dogs. Renal clearance (Cl(re)) of NS-49 was 3.4-fold the glomerular filtration rate (GFR) in the rat in vivo study. The clearance ratio (Cl(re)/GFR) approached unity during cimetidine infusion. Change in the urine flow rate or urinary pH did not affect the Cl(re) of NS-49. The stop-flow patterns of NS-49 in the rabbits and dogs showed a secretion peak in the proximal tubules. On concomitant administration of cimetidine, the secretion peak disappeared, the stop-flow pattern showing neither a secretion nor reabsorption peak. These findings indicate that in these species NS-49 undergoes glomerular filtration and extensive proximal tubular secretion, but little reabsorption. A transport mechanism study of NS-49 in brush-border membrane vesicles (BBMVs) isolated from rat kidney cortex showed that it is transported via the carrier-mediated H(+)/organic cation antiport system. In the rat renal clearance studies (in vivo) tubular secretion of NS-49 was significantly inhibited by quinine (p<0.01) but not by quinidine. Transport studies done with rat BBMVs (in vitro) also showed quinine to be more potent than quinidine in inhibiting NS-49 uptake. These results indicate that stereoselective interaction occurs in active renal tubular secretion.     

The mechanism of the renal excretion of disopyramide in rats. I]

The mechanism involved in the renal excretion of disopyramide (DPM) is still incompletely understood. The purpose of this study was to examine the renal handling of DPM and the interactions between DPM and several organic anionic or cationic drugs related to the renal tubular secretion, using the renal clearance and renal cortical slices uptake techniques in rats. The clearance ratio of DPM was greater than that of glomerular filtration and this suggests the tubular secretion of DPM. The clearance ratio of DPM did not change after infusion of either anionic drugs (p-aminohippurate and probenecid) or a cationic drug (cimetidine). The results of time and concentration-dependent experiments using renal cortical slices demonstrated that DPM was accumulated against a concentration gradient by a saturable process. Inhibition of uptake by 2,4-dinitrophenol and cyanide indicated an energy dependence. DPM uptake was considerably inhibited by the cationic drugs, cimetidine and quinine, suggesting that DPM was transported by the cation transport mechanism. Probenecid, a competitor for the anion transport mechanism, moderately inhibited DPM uptake.     

Transporter-mediated renal handling of nafamostat mesilate

Nafamostat mesilate (NM) is a serine-protease inhibitor that is rapidly eliminated from the circulation and accumulated in the kidney. This study was conducted to characterize the mechanism of NM transport in the kidney because a serious side effect of NM-induced hyperkalemia may be related to accumulation of NM in the kidney. Measurements of uptake of NM in vivo by the kidney uptake index (KUI) method and of transport in an in vitro-cultured LLC-PK1 cell system suggested the involvement of an organic cation transporter (OCT). To clarify the involvement of OCTs located in the basolateral membrane of proximal tubules, we evaluated NM transport by OCTs expressed in Xenopus laevis oocytes. The IC(50) values of NM on [(14)C]TEA ([(14)C]tetraethylammonium) uptake by rOCT1, rOCT2, and hOCT2 were 50, 0.5, and 20 microM, respectively, and NM was concluded to be a substrate of OCTs. To investigate the transport of NM across the brush-border membrane, we examined the uptake of NM into brush-border membrane vesicles (BBMVs) isolated from rat renal cortex. NM was taken up into the BBMVs, and the uptake was decreased by unlabeled NM and temperature, implying that a transporter(s) is also involved in NM transport across the apical membrane. NM was not a substrate of hOCTN1, hOCTN2, or P-gp, implying the involvement of some unknown transporter(s). Thus, renal accumulation of NM can be explained by the involvement of the basolateral OCTs, though the influence of the apical membrane transporter remains to be clarified.     

Characterization of prostaglandin E1 transport in rat renal brush-border membrane

Transport of prostaglandin E(1) (PGE(1)) was investigated in rat renal brush-border membrane vesicles. The uptake of [(3)H]PGE(1) was sensitive to osmosis and temperature. This uptake was saturable and mediated by high-affinity (K(m)=2.1 microM)/low-capacity (V(max)=17.4 pmol/mg protein/30 sec) and low-affinity (K(m)=526.5 microM)/high-capacity (V(max)=1,032.5 pmol/mg protein/30 sec) transport systems. [(3)H]PGE(1) uptake was Na(+)-independent and inhibited by various eicosanoids including PGE(2) and PGF(2alpha). Bromcresol green and sulfobromophthalein, potent inhibitors of prostaglandin transporter (PGT), significantly decreased [(3)H]PGE(1) uptake. Uptake was also inhibited by indomethacin and probenecid, which reportedly have little effect on PGT. Benzylpenicillin and taurocholate decreased the uptake of [(3)H]PGE(1). Like p-[(14)C]aminohippurate (PAH) uptake by vesicles, the uptake of [(3)H]PGE(1) was stimulated by an inside-positive membrane potential, created by applying an inward K(+) gradient and valinomycin. However, the uptake of [(3)H]PGE(1) was not inhibited by PAH, suggesting that PAH and PGE(1) are transported by separate transport systems. [(3)H]PGE(1) uptake was not stimulated by outwardly directed gradients of Cl(-) nor unlabeled PGE(1), indicating that an anion exchanger may not be involved in PGE(1) transport. These findings suggest that the transport of PGE(1) in rat renal brush-border membrane is mediated by specific transport system(s), at least in part, by a potential-sensitive transport system.