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The exit of HCO3- across the basolateral membrane of the proximal tubule cell occurs via the electrogenic cotransport of 3 eq of base per Na+. We have used basolateral membrane vesicles isolated from rabbit renal cortex to identify the ionic species transported via this pathway. Media of varying pH and pCO2 were employed to evaluate the independent effects of HCO3- and CO3(2-) on 22Na transport. Na+ uptake was stimulated when [CO3(2-)] was increased at constant [HCO3-], indicating the existence of a transport site for CO3(2-). In the presence of HCO3-, Na+ influx was stimulated more than 3-fold by an inward SO3(2-) gradient. SO3(2-)-stimulated Na+ influx was stilbene-sensitive, confirming that it occurs via the Na+-HCO3- cotransport system. Na+-SO3(2-) cotransport was demonstrated and found to have a 1:1 stoichiometry. Increasing [CO3(2-)] at constant [HCO3-] reduced the stimulation of Na+ influx by SO3(2-), suggesting competition between SO3(2-) and CO3(2-) at a common divalent anion site. Additional divalent anions that were tested, such as SO4(2-), oxalate2-, and HPO4(2-), did not interact at this site. SO3(2-) stimulation of Na+ influx was absolutely HCO3-(-)dependent and was increased as a function of [HCO3-], indicating the presence of a separate HCO3- site. Lastly, we tested whether Na+ interacts via ion pair formation with CO3(2-) or binds to a distinct site. Na+, which has lower affinity than Li+ for ion pair formation with CO3(2-), was found to have greater than 5-fold higher affinity than Li+ for the Na+-HCO3- cotransport system. Moreover, when its inhibition was studied as a function of [Na+], harmaline was found to be a competitive inhibitor of Na+ influx, indicating the existence of a distinct cation site. Our data are compatible with a model in which base transport across the basolateral membrane of the proximal tubule cell takes place via 1:1:1 cotransport of CO3(2-), HCO3-, and Na+ on distinct sites.

Na+-stimulated H+ movement and H+-stimulated Na+ uptake were studied in basolateral (blLPM) and canalicular (cLPM) rat liver membrane vesicles. H+ movement was monitored with the fluorescent amine acridine orange; 22Na uptake was assayed by a rapid Millipore filtration technique. In blLPM, inwardly directed Na+ gradients stimulated H+ efflux and outwardly directed Na+ gradients stimulated proton influx. Outwardly directed proton gradients (pH in 5.9/pH out 7.9) stimulated initial 22Na uptake rates 5- to 10-fold over pH-equilibrated conditions (pH in 7.9/pH out 7.9). Conversely, inwardly directed proton gradients (pH in 7.9/pH out 5.9) inhibited 22Na uptake. pH-dependent 22Na uptake was inhibited by amiloride and harmaline but not by other transport inhibitors, bumetanide, furosemide, 4,4'-diisothiocyano-2,2'-disulfonic acid stilbene, 4-acetamido-4'-isothiocyanostilbene-2,2'-disulfonic acid, and acetazolamide. Lithium also inhibited H+-stimulated 22Na uptake. Although a component of pH-stimulated 22Na uptake appeared to be dependent on membrane potential, this electrogenic component was amiloride insensitive. Proton gradient-stimulated 22Na uptake in blLPM was saturable, with a Km of 5.4 mM and a Vmax of 14 nmol . min-1 . mg prot-1. In contrast, in cLPM, no Na+ gradient-stimulated proton movement and no pH-dependent Na+ uptake occurred. These findings establish an electroneutral Na-H antiport in blLPM but not cLPM in rat liver. The polarity of this exchanger supports a model of bile formation that is dependent, in part, on canalicular HCO-3 and/or OH- excretion.

We investigated the mechanism of Cl- transport in microvillus membrane vesicles isolated from Necturus kidneys. Cl- influx was insensitive to changes in membrane potential induced by K+ gradients and the K+ ionophore valinomycin, arguing against conductive Cl- transport. Inward gradients of Na+ or Na+ + K+ did not stimulate initial Cl- influx, arguing against direct Na+-Cl- or Na+-K+-Cl- cotransport. External Cl-, HCO3-, and NO3- each stimulated 36Cl efflux and inhibited 36Cl uptake, indicating anion exchange. Outward HCO3- gradients but not OH- gradients stimulated 36Cl influx, consistent with Cl- -HCO3- exchange. Cl- transport via anion exchange was inhibited by furosemide, bumetanide, and disulfonic stilbenes, but not by acetazolamide. External halides stimulated 36Cl efflux (Cl- = Br- greater than I- greater than F-) but the organic anions lactate, p-aminohippurate, and urate did not. Amiloride-sensitive Na+-H+ exchange was demonstrated. Finally, in the presence of a CO2/HCO3 buffer system, imposing an inward Na+ gradient caused a time-delayed stimulation of 36Cl uptake, consistent with indirect coupling of Na+-H+ and Cl- -HCO3- exchangers. We conclude that the parallel operation of Na+-H+ and Cl- -HCO3- exchangers rather than direct cotransport may account for the Na+-coupled uphill Cl- entry previously observed in the intact proximal tubular cell of Necturus.

Chronic K loading and Na deficiency increase the capacity of colonic cells to secrete K and are characterized by increases in Na-K-ATPase activity and, as recently shown by our laboratory, the area of basolateral membrane. The present experiments were performed in rat colon to evaluate the quantitative and qualitative changes that occur in the enzymes after induction of K adaptation. Changes in specific binding of [3H]ouabain to a plasma membrane-rich fraction of colonic mucosal cells correlated directly with changes in enzyme activity. In Na-deficient animals the number of ouabain-binding sites increased from 63.6 in controls to 111.2 pmol/mg protein, while the KD for ouabain was unchanged. The Km for ATP was not altered in experimental animals, although Vmax rose twofold. Further studies showed no change in the activity of adenylate cyclase, a marker enzyme for basolateral membrane, in the plasma membrane-rich fraction from experimental cells, although Na-K-ATPase activity increased significantly. These data indicate that increased Na-K-ATPase activity in K-adapted colonic cells reflects an absolute increase in number of pump sites, due, at least in part, to an increase in area of basolateral cell membrane. In addition, in sodium-deprived animals the present data suggest an increase in the density of K pumps in basolateral cell membrane.

The pathways for transport of Cl- and formate in microvillus membrane vesicles isolated from rabbit renal cortex were evaluated. An outward formate gradient stimulated the uptake of Cl-, and an outward Cl- gradient stimulated the uptake of formate, indicating Cl-/formate exchange. In addition, an inside alkaline pH gradient induced the accumulation of formate, consistent with nonionic diffusion of formic acid. Although an inward Na+ gradient also stimulated uphill formate accumulation, suggesting Na+/formate cotransport, this effect was abolished when ionophores were used to prevent the generation of a transmembrane pH gradient, indicating an indirect coupling of formic acid transport to Na+/H+ exchange. An inside alkaline pH gradient only minimally stimulated the uptake of 82Br-, used as tracer for Cl-, confirming the absence of appreciable Cl-/OH- exchange. However, the same pH gradient in the presence of a physiologic formate concentration (0.2 mM) markedly stimulated 82Br- influx. These data suggest that Cl-/formate exchange with recycling of formic acid by nonionic diffusion is a potential mechanism for active Cl- absorption across the luminal membrane in the proximal tubule and perhaps in other epithelia.

We evaluated the H+ :Na+ coupling ratio of the Na+ -H+ exchanger present in microvillus membrane vesicles isolated from the rabbit renal cortex. Our approach was to impose transmembrane Na+ and H+ gradients of varying magnitude and then to measure the net flux of Na+ over the subsequent 5-s period. The Na+ -H+ exchanger was observed to be at equilibrium (i.e. no significant net Na+ flux) whenever [Na+]i/[Na+]0 was equal to [H+]i/[H+]0. Moreover, under all conditions the magnitude and direction of net Na+ flux was independent of changes in the transmembrane electrical potential difference. These results are consistent with a value of 1.0 for the coupling ratio of Na+ -H+ exchange in renal microvillus membrane vesicles.

A potassium loading study was performed in acutely nephrectomized rats to determine the extrarenal effects of diuretics on potassium tolerance. Four diuretics were evaluated: hydrochlorothiazide, furosemide, bumetanide, and spironolactone. Following an intravenous potassium load (0.17 mEq/100 g over one hour), plasma potassium concentration rose by 2.69 +/- 0.26 to 3.67 +/- 0.20 mEq/L in all groups. There was no difference in the observed increment in plasma potassium concentration between animals receiving diuretics and control animals. These results demonstrate that, at the doses used, diuretics do not impair extrarenal potassium disposal in the rat.

Cl-transport was studied in microvillus membrane vesicles isolated from the rabbit renal cortex. Inwardly directed K+ gradients in the presence of the K+ ionophore valinomycin (Val) enhanced 10 mM 36Cl uptake 2.5-fold, confirming a Cl- conductive pathway. An inwardly directed H+ gradient (pHin 7.5, pHout 6.0) stimulated 10 mM Cl- uptake 1.5-fold compared with pHin = pHout = 6.0. However, this H+ gradient stimulation of Cl- uptake appeared secondary to the H+ diffusion potential rather than to Cl-OH exchange, as it was abolished by Val and K+in = K+out. Additional evidence against Cl- transport via anion exchange was the failure of an inwardly directed Cl- gradient to generate an inside-acid pH gradient as monitored by quenching of acridine orange fluorescence. Cl- influx was the same in the presence of inwardly directed gradients of Na+, K+, Cs+, Li+, and Rb+, arguing against NaCl cotransport. Finally, conductive Cl- transport was reduced by the inhibitors furosemide, 4,4'-diisothiocyanostilbene-2,2'-disulfonic acid and 4-acetamido-4'-isothiocyanostilbene-2,2'-disulfonic acid. These data indicate the presence of an inhibitor-sensitive, conductive mode of Cl- transport but fail to demonstrate significant pathways for Cl- OH exchange or NaCl cotransport in rabbit renal microvillus membrane vesicles.

The effect of amphotericin B on glucose and Na+ transport was studied in the Necturus proximal tubule and in microvillus membrane vesicles isolated from the rabbit renal cortex. In the Necturus experiments, the rate constants for disappearance of radiolabeled glucose (kG) and mannitol (kM) from the tubular lumen were determined by stop-flow microperfusion. Saturability and Na+-dependence of glucose reabsorption was confirmed, since kG was reduced by raising intratubular glucose from 1 to 5 mM or by replacing intratubular Na+ with choline. Neither maneuver affected kM. Intratubular amphotericin B (10 microgram/ml), previously shown to stimulate active Na+ reabsorption in the Necturus proximal tubule, inhibited kG with no effect on kM. In the membrane vesicle preparation, amphotericin inhibited the uphill glucose uptake which results from imposing a NaCl gradient from outside to inside, but had no effect on glucose uptake in either the absence of Na+ or in the presence of Na+ when there was no Na+ gradient. Amphotericin B stimulated the uptake of Na+ by the vesicles. The observed dissociation of glucose and Na+ transport by amphotericin B is consistent with the concept that proximal tubular glucose reabsorption is energized by the luminal membrane Na+ gradient and is not directly linked to active Na+ transport per se.

We examined the effects of external H+ on the kinetics of Na+-H+ exchange in microvillus membrane vesicles isolated from the rabbit renal cortex. The initial rate of Na+ influx into vesicles with internal pH 6.0 was optimal at external pH 8.5 and was progressively inhibited as external pH was reduced to 6.0. A plot of 1/V versus [H+]o was linear and yielded apparent KH = 35 nM (apparent pK 7.5). In vesicles with internal pH 6.0 studied at external pH 7.5 or 6.6, apparent KNa was 13 or 54 mM, Ki for inhibition of Na+ influx by external Li+ was 1.2 or 5.2 mM, Ki for inhibition by external NH4+ was 11 or 50 mM, and Ki for inhibition by external amiloride was 7 or 25 microM, respectively. These findings were consistent with competition between each cation and H+ at a site with apparent pK 7.3-7.5. Lastly, stimulation of 22Na efflux by external Na+ (i.e. Na+-Na+ exchange) was inhibited as external pH was reduced from 7.5 to 6.0, also consistent with competition between external H+ and external Na+. Thus, in contrast with internal H+, which interacts at both transport and activator sites, external H+ interacts with the renal microvillus membrane Na+-H+ exchanger at a single site, namely the external transport site, where H+, Na+, Li+, NH4+, and amiloride all compete for binding.

Ionophore use an illustrated by a description of experiments in which the K+ ionophore valinomycin and the uncoupler, carbonyl cyanide p-trifluoromethoxyphenylhydrazone (FCCP) were employed to investigate Na+ transport pathways in renal microvillus membrane vesicles. First, the potential-dependence of solute transport was examined by using valinomycin and K+ gradients to alter the membrane potential. Whereas Na+-glucose cotransport was voltage-sensitive, the transport of Na+ in the absence of glucose was found to occur via a voltage-insensitive process, likely Na+-H+ exchange. Second, FCCP short-circuiting of the membrane vesicles was used to examine possible electrical interactions among Na+ transport pathways. The alanine inhibition of Na+-dependent glucose transport was abolished by FCCP, indicating that the effect of the amino acid on sugar flux was indirect and mediated by an alteration in the membrane potential. Third, aspects of the molecular mechanism of Na+-glucose cotransport were evaluated by using ionophores to study the potential-dependence of phlorizin binding to the Na+-coupled sugar carrier. Whereas the rate of phlorizin association was potential-dependent, the rate of release of bound phlorizin was insensitive to variation in the transmembrane voltage, suggesting that the potential-dependence of Na+-glucose cotransport arises from potential-dependent behavior of the free carrier rather than from potential-dependence of Na+-glucose translocation per se. These studies demonstrate that the use of ionophores augments the value of employing isolated plasma membrane vesicles to investigate mechanisms of epithelial solute transport.

The major pathway for HCO3- transport across the basolateral membrane of the proximal tubule cell is electrogenic Na+-HCO3- cotransport. In this study, we have determined the stoichiometry of the Na+-HCO3- cotransport system in basolateral membrane vesicles that were isolated from rabbit renal cortex by Percoll gradient centrifugation. When the membrane potential is approximated by the Nernst potential for K+, as in the presence of the K+ ionophore valinomycin, equilibrium thermodynamics predicts that the Na+-HCO3- cotransport system should come to equilibrium and mediate no net flux when (Na)i/(Na)o = [(HCO3)o/(HCO3)i]n[(K)o/(K)i]n-1, where n is the HCO3-:Na+ stoichiometry. Our experimental approach was to impose transmembrane Na+, HCO3-, and K+ gradients of varying magnitude and direction, and then to measure the net flux of Na+ over the subsequent 3-s period. In this way, we could determine the conditions for equilibrium of the transport system and thereby calculate n. The results of these experiments indicate that the value of n is greater than 2.6 and less than 3.5, consistent with a stoichiometry of 3 HCO3-:1 Na+, or a thermodynamically equivalent process. Based on reported intracellular potentials and ion activities, this value for the stoichiometry indicates that the inside-negative membrane potential is sufficient to drive HCO3- exit against the inward concentration gradients of HCO3- and Na+ that are present across the basolateral membrane of the intact proximal tubule cell under physiologic conditions.

Studies on microvillus membrane from rabbit kidney cortex suggest that chloride absorption may occur by chloride/formate exchange with recycling of formic acid by nonionic diffusion. We tested whether this transport mechanism participates in active NaCl reabsorption in the rabbit proximal tubule. In proximal tubule S2 segments perfused with low HCO-3 solutions, the addition of formate (0.25-0.5 mM) to the lumen and the bath increased volume reabsorption (JV) by 60%; the transepithelial potential difference remained unchanged. The effect of formate on JV was completely reversible and was inhibited both by ouabain and by luminal 4,4'-diisothiocyanostilbene-2,2'-disulfonate. Formate (0.5 mM) failed to stimulate JV in early proximal convoluted tubules perfused with high HCO-3 solutions. As measured by miniature glass pH microelectrodes, this lack of formate effect on JV was related to a less extensive acidification of the tubule fluid when high HCO-3 solutions were used as perfusate. These data suggest that chloride/formate exchange with recycling of formic acid by nonionic diffusion represents a mechanism for active, electroneutral NaCl reabsorption in the proximal tubule.

Present evidence suggests that in the small intestine, villus cells are primarily absorptive and crypt cells are primarily secretory. In order to further confirm that there are differences in transport properties between villus and crypt cells, we have separated villus from crypt cells, using calcium chelations techniques, and determined the distribution of Na:H and Cl:HCO3 exchange activity on brush border membrane and basolateral membrane preparations from these two cell populations. Separation of cells was determined utilizing alkaline phosphatase and maltase activity as a marker of villus cells and thymidine kinase activity as a marker of crypt cells. Utilizing these techniques, we were able to sequentially collect cells along the villus-crypt axis. Na-stimulated glucose and alanine uptake in brush border membrane vesicles diminished from the villus to the crypt region in the sequentially collected cells fractions, further suggesting separation of these cells. Brush border and basolateral membranes were then prepared from cells from the villus and crypt areas, utilizing a continuous sucrose gradient. In the villus cells, Na:H exchange activity was found associated with both the brush border and basolateral membrane, whereas, in crypt cells, Na:H exchange activity was only found on the basolateral membrane. Cl:HCO3 exchange activity was found only on the brush border membrane, in both villus and crypt cells. These studies suggest functional heterogeneity in ion transport between villus and crypt cells.

Transport interactions among phosphate, glucose, and alanine were evaluated in brush border membrane vesicles isolated from the rabbit renal cortex. Both glucose and alanine inhibited the Na+ gradient-stimulated uphill accumulation of phosphate. Neither glucose nor alanine inhibited the phosphate uptake measured in the absence of Na+ or in the presence of a collapsed Na+ gradient shunted with amphotericin B. Phosphate and alanine inhibited the Na+ gradient-stimulated uphill accumulation of glucose but not the glucose uptake measured in the presence of a collapsed Na+ gradient shunted with amphotericin B. Alanine, which was more potent than glucose or phosphate in inhibiting Na+ gradient-stimulated solute transport, was also more potent than glucose or phosphate in stimulating the uptake of Na+ into the vesicles. We conclude that the inhibitory interactions among phosphate, glucose, and alanine probably represent indirect effects resulting from solute-induced alterations in the transmembrane electrochemical Na+ gradient rather than direct effects resulting from competition for a polyfunctional carrier or from allosteric interactions. Such a mechanism could explain the inhibitory interactions among phosphate, glucose, and alanine that have been observed in the intact proximal tubule.

The plasma membrane Na+-H+ exchanger is a ubiquitous transport system that participates in diverse cell functions involving the cellular uptake of Na+ or extrusion of H+. It has a tightly coupled 1:1 stoichiometry, has affinity for Li+ and NH+4 in addition to Na+ and H+, and can function in multiple amiloride-sensitive exchange modes involving these cations. These general transport properties may be explained by kinetic models involving either cation-hydroxyl cotransport or actual cation-proton exchange. The most important kinetic property of the Na+-H+ exchanger is its greater than first-order dependence on [H+]i. This property enables the Na+-H+ exchanger to play an important role in the regulation of intracellular pH.

The transport of urate was evaluated in brush border membrane vesicles isolated from the dog renal cortex. It was previously shown that uphill urate and p-aminohippurate (PAH) uptake into these membrane vesicles results from imposing an outwardly directed OH- gradient. In the present study, the OH- gradient-stimulated uptake of urate was inhibited by external Cl-. In the absence of OH- gradients, imposing an outwardly directed Cl- gradient induced the transient accumulation of urate against its concentration gradient (overshoot), whereas imposing an inwardly directed Cl- gradient induced the transient uphill efflux of urate (undershoot). The effects of Cl- gradients on urate transport persisted when the diffusion potentials caused by the Cl- gradients were negated by superimposing K+ gradients in the presence of the K+ ionophore valinomycin. The dose-response curves for three different inhibitors were identical for the OH- and Cl- gradient-stimulated modes of urate uptake. The uptake of PAH was also stimulated by imposing an outwardly directed Cl- gradient. The dose-response curves for probenecid inhibition were identical for the OH- and Cl- gradient-stimulated modes of PAH uptake. Finally, the existence of a Na+ cotransport pathway for urate or PAH could not be demonstrated. We conclude that the principal mechanism for urate transport in dog renal microvillus membrane vesicles is via an anion exchanger with affinity for urate, PAH, OH-, and Cl-. This anion exchanger may play an important role in mediating organic anion reabsorption and secretion in the proximal tubule.

We examined the effect of histidine-specific reagents on the transport activity of the Na+-H+ exchanger in microvillus (brush-border) membrane vesicles isolated from the rabbit renal cortex. Rose bengal-catalyzed photo-oxidation caused irreversible inhibition of the rate of Na+-H+ exchange but also caused significant loss of vesicle integrity. Treatment of the membrane vesicles with diethylpyrocarbonate caused inactivation of Na+-H+ exchange that could not be attributed to vesicle disruption or collapse of transmembrane H+ gradients. Inactivation of Na+-H+ exchange by diethylpyrocarbonate followed pseudo-first order kinetics to below 10% residual activity, could be reversed by hydroxylamine, was reflected by a decreased Vmax with no change in the Km for Na+, was dependent on external pH but not internal pH, was blocked by amiloride, and was enhanced by Na+. These data are consistent with the hypothesis that a diethylpyrocarbonate-sensitive imidazolium residue is the titratable group found in kinetic studies to bind H+ at the external transport site of the Na+-H+ exchanger.

The effects of the hallucinogen harmaline on D-glucose, L-alanine, and Na+ transport were studied in microvillus membrane vesicles isolated from the rabbit renal cortex. Harmaline had no effect on glucose transport in the absence of Na+, but reversibly inhibited sugar flux in the presence of NaCl. Inhibition of Na+-dependent glucose transport was inversely related to the Na+ concentrations. The hallucinogen competitively inhibited the Na+ activation of phlorizin binding to the membranes but did not inhibit phlorizin binding in the absence of Na+. Harmaline inhibited Na+-dependent alanine transport and, at higher drug concentrations, the amino acid flux in the absence of NaCl. Harmaline competitively inhibited the rate of Na+ uptake which, in the absence of glucose and alanine, is known to occur via Na+-H+ exchange. The hallucinogen trans-inhibited the efflux of glucoe and Na+ from membrane vesicles preloaded with the solutes. These findings suggest that harmaline is a direct inhibitor of microvillus membrane transport processes and acts as a competitive inhibitor of Na+ transport sites. Harmaline may therefore be a useful investigative tool for studying mechanisms of Na+-coupled transport in the luminal membrane of the proximal tubular cell.

We evaluated the effects of acetazolamide on Na+-HCO3- cotransport in basolateral membrane vesicles isolated from the rabbit renal cortex. Na+ uptake stimulated by an imposed inward HCO3- gradient was not significantly reduced by 1.2 mM acetazolamide, indicating that acetazolamide does not directly inhibit Na+-HCO3- cotransport. 4,4'-Diisothiocyanostilbene-2,2'-disulfonate (DIDS)-sensitive Na+-base cotransport was found to be absolutely CO2/HCO3--dependent. We therefore tested whether acetazolamide-sensitive availability of HCO3- at the basolateral membrane could be rate-limiting for Na+-base cotransport under some conditions. In the presence of a CO2/HCO3- buffer system but absence of an initial HCO3- gradient, Na+ influx was stimulated fivefold by an outward NH4+ gradient. This stimulation of Na+ influx by an outward NH4+ gradient was inhibited greater than 75% by 0.6 mM acetazolamide, suggesting that acetazolamide blocked the ability of the NH4+ gradient to generate an inward HCO3- gradient. In the presence of an inward HCO3- gradient, Na+ influx was inhibited greater than 70% by an inward NH4+ gradient. This inhibition of Na+ influx was reduced to only 35% by 0.6 mM acetazolamide, suggesting that acetazolamide blocked the ability of NH4+ to collapse the inward HCO3- gradient. Similarly, Na+ influx in the presence of an inward HCO3- gradient was inhibited greater than 80% by an outward acetate gradient, and this inhibition was reduced to only 50% by acetazolamide. Thus, acetazolamide caused either inhibition or stimulation of Na+ uptake depending on the conditions with respect to pH and HCO3- gradients. The indirect interaction of acetazolamide with the basolateral membrane Na+-HCO3- cotransport system may be an important mechanism underlying inhibition of proximal tubule acid secretion by this agent.

Possible transport mechanisms of chloride across the mammalian proximal tubule include both active and passive components and, accordingly, transcellular and intercellular transport routes. Experiments are described in which the possibility of electroneutral anion exchange across the apical cell membrane of proximal tubule cells is evaluated. Experiments in brushborder vesicles of the rabbit kidney cortex have shown the existence of a chloride/formate exchange process. In addition, evidence is presented that formic acid is crossing the brushborder membrane of rabbit renal cortex by non-ionic diffusion. A transport schema is developed in which formate/chloride exchange and sodium/hydrogen exchange in parallel with recycling of formic acid by non-ionic diffusion can account for the electroneutral transport of sodium chloride. Perfusion studies in isolated rabbit straight and convoluted tubules show significant transport stimulation by submillimolar concentrations of formate. Thus, chloride/formate exchange is a possible mechanism for active, transcellular chloride movement across the mammalian proximal tubule.