Effect of recombinant human erythropoietin on the rate of Na+,H+-exchange in erythrocytes from patients with chronic renal failure on hemodialysis

The study explores the effect of recombinant human erythropoietin and the rate of Na⁺,H⁺-exchange in erythrocytes from patients with chronic renal failure undergoing hemodialysis. The rate of Na⁺,H⁺-exchange in erythrocytes from patients was higher than in the control and remained unchanged after 24 months of treatment with erythropoietin. Therapy with recombinant human erythropoietin does not normalize the Na⁺,H⁺-exchange mechanism. It is concluded that factors underlying disturbances of ion transport in erythrocytes from uremic patients cannot be corrected with erythropoietin. Translated fromByulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 124, No. 12, pp. 613–615, December, 1997     

Effect of sarcolemmal anion transport system blockers on the development of ischemia-induced myocardial edema and recovery of contractile function during reperfusion

Subtotal 30-min ischemia leads to myoglobin release and increases water content in the heart. Reperfusion partially restores the developed pressure. Addition of furosemide (a Na⁺, K⁺, 2Cl⁻-sumport blocker) or NMA (inhibitor of Na⁺/H⁺-exchange) to perfusate decreases myocardial water content, reduces myoglobin loss, and completely restores myocardial contractile function. The low-rate perfusion of isolated heart and its reperfusion with solutions containing DIOA (inhibitor of K⁺, Cl⁻-co-transport) or IAA-94 (Cl⁻ channel blocker) increases water accumulation and myoglobin release from the myocardium, and deteriorated its contractile function during reperfusion. Translated fromByulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 127, No. 4, pp. 400–403, April, 1999     

Effect of human defensins on the cytoplasmic Ca2+ content in platelets

The effect of the total fraction of human defensins (HNP-1, HNP-2, and HNP-3) on the cytoplasmic Ca²⁺ content ([Ca²⁺]_i) in the platelets of healthy donors was studied. At concentrations of 0.1–40 μg/ml and an incubation time of 10 min defensins have no effect on [Ca²⁺]_i in platelets labeled with Fura-2AM. However, at higher concentrations (100 μg/ml) they increased platelet [Ca²⁺]_i. In addition, defensins (40 μg/ml) inhibited the Ca²⁺ increase in platelets induced by thrombin, adenosine diphosphate, and the lipopolysaccharide ofS. typhimurium endotoxin. The most pronounced inhibitory effect was observed in a suspension of thrombin-stimulated platelets. It is shown that the effect of human defensins on the functional activity of platelets is due to the alterations in the intracellular Ca²⁺. Translated fromByulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 118, N ^o 12, pp. 600–603, December, 1994     

Role of ionic transport in regulation of hemoglobin affinity for oxygen in diabetes mellitus

The rate of Na⁺/H⁺ exchange is increased by 24%, activities of Ca-dependent K+ channels is increased by 13%, and activity of erythrocyte Ca²⁺-ATPase decreased by 17% in patients with diabetes mellitus concomitant with essential hypertension in comparison with patients with essential hypertension without disorders of carbohydrate metabolism. Changes in activity of Na⁺/H⁺ exchange, Ca-dependent K⁺ channels, and erythrocyte Ca²⁺-ATPase and increased oxygen affinity of hemoglobin are due to increased glucose concentration in the plasma and are leveled by olifen.     

Mobilization of intracellular Ca2+ induced by ADP and thrombin in platelets from diabetic patients with vascular complications

It is shown that the baseline level of cytoplasmic Ca²⁺ in platelets from diabetic patients is nearly 1.5 times as high as in healthy donors. The thrombin-induced increase of intracellular Ca²⁺ in patients with angiopathies is reliably lower than in the control, while in patients without angiopathies it is higher than that in donor platelets. The evevation of cytoplasmic Ca²⁺ induced by ADP is greater in both groups of patients. No Changes are found in the baseline level of intracellular Ca²⁺ or in the ADP-induced concentration of Ca²⁺ in platelets from diabetic patients during a 12-week course of insulin therapy. The intracellular Ca²⁺ does not rise after 2 weeks of insulin treatment in platelets from diabetic patients. Translated fromByulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 121, N ^o 1, pp. 112–115, January, 1996     

Regulation of Na+/H+ exchange in opossum kidney cells by parathyroid hormone,cyclic AMP and phorbol esters

Parathyroid hormone (PTH) controls two proximal tubular brush border membrane transport systems, Na⁺/phosphate co-transport and Na⁺/H⁺ exchange. In OK cells, a cell line with proximal tubular transport characteristics, PTH acts via kinase C and kinase A activation to inhibit Na⁺/phosphate co-transport [6, 8, 9, 19, 22]. In the present study, we show that PTH inhibits Na⁺/H⁺ exchange and that this effect can be mimicked by pharmacological activation of kinase A and kinase C. Ionomycin-dependent increases in cytoplasmic Ca²⁺ concentration do not induce inhibition of Na⁺/H⁺ exchange; PTH-dependent inhibition of Na⁺/H⁺ exchange is not prevented by ionomycin or by the intracellular Ca²⁺ chelator 1,2-bis(2-aminophenoxy)ethane-N,N,N,N-tetraacetic acid (Ca²⁺ clamping). Detailed dose-response curves for the different agonists, given either alone or in combination, suggest that the two regulatory cascades (kinase A and kinase C) are operating independent of each other and reach a common final target, resulting in 40–50% inhibition of Na⁺/H⁺ exchange. An analysis of intracellular pH sensitivity of Na⁺/H⁺ exchange suggests that inhibition is not related to a shift in set point, but is rather explained by a reduced V _max of Na⁺/H⁺ exchange and/or reduced affinity for protons at the internal membrane surface. It is suggested that kinase A as well as kinase C can mediate PTH inhibition of renal proximal tubular Na⁺/H⁺ exchange and that the relative importance of a particular regulatory cascade is determined by the PTH-concentration-dependent rates in the liberation of diacylglycerol (phospholipase C/kinase C) and cAMP (adenylate cyclase/kinase A).Abbreviations HEPES 4-(2-hydroxyethyl)-1-piperazineethane-sulphonic acid - EDTA ethylenediaminetetraacetic acid - FCS fetal calf serum - PTH parathyroid hormone - Tris 2-amino-2-hydroxyme-thylpropane-1,3-diol - BCECF 2,7-bis(2-carboxyethyl)-5,6-carboxyfluorescein - P_i inorganic phosphate - pH_i intracellular pH - Ca_i cytosolic free Ca²⁺ - BAPTA 1,2-bis(2-aminophenoxy)ethane-N,N,N,N-tetraacetic acid - EGTA ethylene glycol-bis(-amino-ethylether)-N,N,N,N-tetraacetic acid - IP₃ inositol 1,4,5-trisphos-phate - DAG 1,2-diacylglycerol     

A study of the mechanism of action of befol on Ca2+ metabolism in cardiomyocytes using a FURA-2 fluorescent probe

The dynamics of the Ca-response of cardiomyocytes is studied and the efficiency of befol, verapamil, and amiodarone is compared using various experimental models of stimulation of [Ca²⁺]_i. Befol (1–5 μM) is shown to inhibit the caffeine-and strophanthin G-induced rise of [Ca²⁺]_i. Unlike verapamil and amiodarone, befol exhibits no Ca-blocking activity in modeled K-depolarization. It is concluded that the cardiotropic effect of befol is mediated through its primary action on Na⁺/Ca²⁺ exchange in cardiomyocytes, while the cardioplegic effect of verapamil and amiodarone is due to their ability to block the slow Ca²⁺ inward current. Translated fromByulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 121, N ^o 3, pp. 288–291, March, 1996     

Regulation of the cytosolic pH set point for activation of the Na+/H+ antiport in human platelets: the roles of the Na+/Ca2+ exchange,the Na+-K+-2Cl− cotransport and cellular volume

To explore further the mechanisms that regulate the Na⁺/H⁺ antiport in human platelets, we examined the effect of Na⁺ pump inhibition by ouabain and K⁺ removal from the extracellular medium on parameters of this transport system. Treatment with ouabain resulted in increased cytosolic free Ca²⁺ and Na⁺, coupled with an alkaline shift in the cytosolic pH set point for the Na⁺/ H⁺ antiport. Inhibition of the Na⁺ pump by the removal of K⁺ from the medium increased the cytosolic Na⁺ but not the cytosolic Ca²⁺; yet this treatment also produced a substantial alkaline shift in the cytosolic pH set point for the Na⁺/H⁺ antiport. This effect appeared to relate to a decline in cellular volume and it was attenuated by the Na⁺-K⁺-2Cl^– cotransport inhibitor, bumetanide. These findings indicate: (a) a link between the Na⁺ pump and the Na⁺/H⁺ antiport, mediated by the Na⁺/Ca²⁺ exchange and the cytosolic free Ca²⁺, and (b) a link between the Na⁺/H⁺ antiport and the Na⁺-K⁺-2Cl^– cotransport through cellular volume.This work was supported by grants from the National Heart, Lung, and Blood Institute (HL34807, HL42856) and the American Diabetes Association. M. Kimura is a postdoctoral research fellow of the American Heart Association, New Jersey Affiliate     

Thrombin-induced cytosolic alkalinization in human platelets occurs without an apparent involvement of HCO 3 − /Cl− exchange

We have estimated the changes in cytosolic pH (pH_i) that occur when human platelets are stimulated by thrombin. Changes in pH_i were estimated (i) from the H⁺ efflux across the plasma membrane using an extracellular pH electrode and (ii) using an intracellular pH-sensitive fluorescent dye (BCECF). Stimulation of platelets with thrombin (0.5 unit/ml) resulted in an H⁺ efflux that averaged 7.7±1.6 mol/10¹¹ platelets (means±SD) leading to an increase in pH_i, from 7.05±0.04 to 7.45±0.05. Both H⁺ efflux and pH_i changes were unaffected by 0.1 mM 4,4-diisothiocyanostilbene-2,2 disulphonate (DIDS), 0.1 mM 4-acetamido 4-isothiostilbene-2,2-disulphonic acid (SITS), or 0.5 mM bumetanide, suggesting no involvement of anion transport systems, e.g. an HCO ₃ ^– /Cl^– exchange. Removal of HCO ₃ ^– or Cl^– from the suspending buffer had no effect on the extent of the rise in pH_i. After blockade of Na⁺/H⁺ exchange by 100 M ethylisopropylamiloride (EIPA), thrombin induced a decrease in pH_i the rate of which averaged 0.39 unit/min in HCO ₃ ^– -containing medium, and 0.57 unit/min in HCO ₃ ^– -free medium. The cytosolic buffer capacity for H⁺ was determined by the nigericin/ NH₄Cl technique in BCECF-loaded platelets and averaged 25.3 mmol/(1xpH) in buffer containing 8 mM HCO ₃ ^– , but only 17.2 mmol/(1xpH) in HCO ₃ ^– -free buffer. The total amount of H⁺ transferred by Na⁺/H⁺ exchange can be estimated from our measurements at 10 mmol/l platelet cytosol in the absence of HCO ₃ ^– and to 14 mmol/l platelet cytosol in the presence of HCO ₃ ^– , and is in good agreement with the estimated amount of Na⁺ uptake by ADP-stimulated platelets. We conclude that net extrusion of H⁺ from stimulated platelets is predominantly mediated by Na⁺/H⁺ exchange without an apparent contribution of HCO ₃ ^– /Cl^– exchange.     

Short-term and long-term stimulation of Na+−H+ exchange in cortical brush-border membranes during compensatory growth of the rat kidney

The effect of unilateral nephrectomy on Na⁺–H⁺ exchange in rat renal cortical brush-border membrane vesicles (BBMV) was studied by the method of acridine orange fluorescence quenching. The exchanger activity in BBMV from remnant kidney increased rapidly by 70–75% within first 30 min following uninephrectomy. Only a slight further increase was found in later stages of renal growth, i.e. 30 min to 7 days following uninephrectomy. The changes in antiporter activity were restricted toV _max, whereas theK _m for Na⁺ was similar in control and compensatory growing kidney. The increase of Na⁺–H⁺ exchange at 15 min was not affected by actinomycin D in vivo, whereas the increase at 48 h was completely abolished indicating that protein synthesis could be involved in the late, but not in the initial stimulation of renal Na⁺–H⁺ exchange. The late, but not the initial stimulations of Na⁺–H⁺ exchange were associated with elevated activities of cortical (Na⁺+K⁺)-ATPase indicating that changes in antiporter activity precede those in the (Na⁺+K⁺)-pump. The early stimulation of Na⁺–H⁺ exchange in BBMV in one kidney was induced also by the occlusion of blood flow through the contralateral kidney for 15 min, without removing it. Thirty min after the occlusion was removed and the reflow established, the Na⁺–H⁺ exchange in BBMV from the intact kidney decreased to the control values. The observed modulations in renal Na⁺–H⁺ exchanger may be regulated by phosphorylation-dephosphorylation events. In support, the concentration of a well known protein kinase C activator, 1,2-diacylglycerol, in the cortical tissue of the remnant kidney increased up to 100% within 5 min following unilateral nephrectomy and preceded the increase in Na⁺–H⁺ exchange. The early stimulation of Na⁺–H⁺ exchange may be a trigger in initiating the kidney growth.     

Effects of the nootropics piracetam and GVS-111 on voltage-dependent ion channels of neuronal membranes

The effect of two nootropics, piracetam and N-phenylacetyl-L-prolyglycine ethyl ester (GVS-111), is studied by measuring high-threshold K⁺ and Ca²⁺ currents in isolated snail neurons using a two-microelectrode patch-clamp technique. Piracetam and GVS-111 are shown to reduce the amplitude of both the K⁺ and the Ca²⁺ (to a lesser extent) current. The threshold concentrations for GVS-111 and piracetam are 10⁻⁹-10⁻⁸ M and 1–5×10⁻⁴ M, respectively. It is assumed that the antiamnestic effect of the nootropics is partially mediated by a blockade of ion channels of the neuronal membrane. Translated fromByulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 121, N ^o 2, pp. 151–155, February, 1996 Presented by G. N. Kryzhanovskii, Member of the Russian Academy of Medical Sciences     

Intracellular Na+ modulates large conductance Ca2+-activated K+ currents in human umbilical vein endothelial cells

We studied the effects of Na⁺ influx on large-conductance Ca²⁺-activated K⁺ (BK_Ca) channels in cultured human umbilical vein endothelial cells (HUVECs) by means of patch clamp and SBFI microfluorescence measurements. In current-clamped HUVECs, extracellular Na⁺ replacement by NMDG⁺ or mannitol hyperpolarized cells. In voltage-clamped HUVECs, changing membrane potential from 0 mV to negative potentials increased intracellular Na⁺ concentration ([Na⁺]_i) and vice versa. In addition, extracellular Na⁺ depletion decreased [Na⁺]_i. In voltage-clamped cells, BK_Ca currents were markedly increased by extracellular Na⁺ depletion. In inside-out patches, increasing [Na⁺]_i from 0 to 20 or 40 mM reduced single channel conductance but not open probability (NPo) of BK_Ca channels and decreasing intracellular K⁺ concentration ([K⁺]_i) gradually from 140 to 70 mM reduced both single channel conductance and NPo. Furthermore, increasing [Na⁺]_i gradually from 0 to 70 mM, by replacing K⁺, markedly reduced single channel conductance and NPo. The Na⁺–Ca²⁺ exchange blocker Ni²⁺ or KB-R7943 decreased [Na⁺]_i and increased BK_Ca currents simultaneously, and the Na⁺ ionophore monensin completely inhibited BK_Ca currents. BK_Ca currents were significantly augmented by increasing extracellular K⁺ concentration ([K⁺]_o) from 6 to 12 mM and significantly reduced by decreasing [K⁺]_o from 12 or 6 to 0 mM or applying the Na⁺–K⁺ pump inhibitor ouabain. These results suggest that intracellular Na⁺ inhibit single channel conductance of BK_Ca channels and that intracellular K⁺ increases single channel conductance and NPo. GH Liang and MY Kim contributed equally to this publication and therefore share the first authorship.     

Potassium and sodium ions in the nerve tissue of emotionally overstrained rabbits

Measurement of K⁺ and Na⁺ concentrations in samples of individual brain nuclei and in ganglia of the autonomic nervous system from rabbits subjected to severe emotional stress (ES) through aperiodic stimulation of ventromedial hypothalamic nuclei and electrocutaneous stimulation revealed significantly altered levels of these ions in locus ceruleus samples from animals predisposed to ES-induced cardiovascular disorders and in samples of neurons of the caudal part of the brainstem from those resistant to such disorders. Translated fromByulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 118, No 8, pp. 129–131, August, 1994 Presented by K. V. Sudakov, Member of the Russian Academy of Medical Sciences     

Drug Modulation of Transductor Function of Na+,K+-ATPase

Experimental proof of the hypothesis on modulation of the transductor function of Na⁺,K⁺-ATPase by the ouabain—Ca²⁺ chelate complex was obtained by the method organotypic tissue culture. Quantum chemical estimations detected two principally different modes of Ca²⁺ ion chelation by ouabain molecule. It is hypothesized that the ouabain—Ca²⁺—Na⁺,K⁺-ATPase ligand-receptor complex is formed due to ion-ion bonds. The formation of the complex serves as the signal triggering the enzyme transductor function. It is experimentally proven that ouabain is incapable of inhibiting neurite growth in sensory neuron and heart tissue explants after removal of free calcium from the nutrient medium with EGTA. Translated from Byulleten’ Eksperimental’noi Biologii i Meditsiny, Vol. 146, No. 10, pp. 416–418, October, 2008     

Aldosterone actions on basolateral Na+/H+ exchange in Madin-Darby canine kidney cells

In recent studies, there has been a re-evaluation of the polarity of Na⁺/H⁺ exchange in Madin-Darby canine kidney (MDCK) cells. This study was designed to examine aldosterone actions on basolaterally located Na⁺/H⁺ exchange of MDCK cell monolayers grown on permeant filter supports; pH_i was analysed in the absence of bicarbonate by using the pH-sensitive fluorescent probe 2,7-bis(carboxyethyl)-5,6-carboxyfluorescein. Pre-exposure of MDCK cells to aldosterone led within 10–20 min to an alkalization of pH_i ( 0.3 pH unit); this effect is prevented by an addition of dimethylamiloride to the basolateral superfusate. Addition of aldosterone led to stimulation of the basolaterally located Na⁺/H⁺ exchange activity (Na⁺-dependent recovery from an acid load); this effect required preincubation (more then 3 min) and was observed at 0.1 nM aldosterone. Preexposure (15 min) of MDCK monolayers to phorbol 12-myristate 13-acetate also led to an activation of Na⁺/H⁺ exchange; pre-exposure to 8-bromo-cAMP led to inhibition of Na⁺/H⁺ exchange activity. An inhibitory effect of aldosterone was observed if Na⁺/H⁺ exchange activity was analysed in the presence of aldosterone; the highest inhibitory effects (20%–30%) occurred at concentrations of 5 nM and higher. Aldosterone-dependent inhibition does not require preincubation and is fully reversible; it was only observed at low (20 mM) but not at high Na⁺ concentrations (130 mM). The data suggest that aldosterone has an instantaneous inhibitory effect on basolaterally located Na⁺/H⁺ exchange activity under conditions of low Na⁺, but stimulates the rate of transport activity upon preincubation under conditions of physiological Na⁺ concentrations.     

Changes in rhythmoinotropic reactions of the myocardium in chronic ischemia: Pathology or adaptation?

Changes in the organization of the Ca⁺-transporting systems but not disturbances in the contractile apparatus of cardiomyocytes are shown to occur in chronic coronary heart disease. During a certain stage of CHD the change in rhythmoinotropic relations may reflect adaptive changes induced by functioning of the myocardium under conditions of ischemia. Translated fromByulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 117, N ^o 5, pp. 457–459, May, 1994     

The rise of [Na+]i during ischemia and reperfusion in the rat heart—underlying mechanisms

Intracellular Na⁺ concentration ([Na⁺]_i) rises in the heart during ischemia, and on reperfusion, there is a transient rise followed by a return toward control. These changes in [Na⁺]_i contribute to ischemic and reperfusion damage through their effects on Ca²⁺ overload. Part of the rise of [Na⁺]_i during ischemia may be caused by increased activity of the cardiac Na⁺/H⁺ exchanger (NHE1), activated by the ischemic rise in [H⁺]_i. In support of this view, NHE1 inhibitors reduce the [Na⁺]_i rise during ischemia. Another possibility is that the rise of [Na⁺]_i during ischemia is caused by Na⁺ influx through channels. We have reexamined these issues by use of two different NHE1 inhibitors, amiloride, and zoniporide, in addition to tetrodotoxin (TTX), which blocks voltage-sensitive Na⁺ channels. All three drugs produced cardioprotection after ischemia, but amiloride (100 μM) and TTX (300 nM) prevented the rise in [Na⁺]_i during ischemia, whereas zoniporide (100 nM) did not. Both amiloride and zoniporide prevented the rise of [Na⁺]_i on reperfusion, whereas TTX was without effect. In an attempt to explain these differences, we measured the ability of the three drugs to block Na⁺ currents. At the concentrations used, TTX reduced the transient Na⁺ current (I _Na) by 11 ± 2% while amiloride and zoniporide were without effect. In contrast, TTX largely eliminated the persistent Na⁺ current (I _Na,P) and amiloride was equally effective, whereas zoniporide had a substantially smaller effect reducing I _Na,P to 41 ± 8%. These results suggest that part of the effect of NHE1 inhibitors on the [Na⁺]_i during ischemia is by blockade of I _Na,P. The fact that a low concentration of TTX eliminated the rise of [Na⁺]_i during ischemia suggests that I _Na,P is a major source of Na⁺ influx in this model of ischemia.     

Potassium transport through the erythrocyte plasma membrane in patients with insulin-dependent diabetes mellitus: Effects of insulin therapy

Plasma membrane potential and function of Ca²⁺-activated K⁺ channels of erythrocytes are studied in patients with insulin-dependent diabetes mellitus. A significant increase in membrane potential and in the degree of opening of Ca²⁺-activated K⁺ channels is revealed in comparison with erythrocytes of healthy donors. The degree of channel opening is higher in patients with nephropathy than in those with retinopathy, in which this parameter is normal. Insulin therapy normalize the erythrocyte plasma membrane potential in the patients and has no effect on the degree of channel opening. Ion transport disturbances and modulations of Na⁺, K⁺, and Ca²⁺ levels in erythrocytes may impair their function in insulin-dependent diabetes mellitus. Translated fromByulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 122, No. 7, pp. 109–113, July, 1996     

Electrical and molecular coupling between sodium and proton fluxes in basolateral membrane vesicles of rat liver

Mechanisms of Na⁺–H⁺ exchange in the hepatocyte were studied utilizing isolated basolateral membrane vesicles prepared by two different methods: Evidence was obtained for the existence of molecular coupling of Na⁺ and H⁺ fluxes (Na⁺/H⁺-antiport) which exhibits saturation kinetics (Km 7 mmol/l Na⁺) and is inhibited by amiloride (1.0 mmol/l). Although the two membrane preparations showed differences with respect to ionic permeabilities, our data suggest that a relatively high H⁺ conductance exists in the basolateral plasma membrane. Hence, electrical coupling of conductive H⁺ and Na⁺ fluxes in the opposite direction could contribute to net Na⁺–H⁺ exchange across the basolateral hepatocyte plasma membrane.     

Mechanism of hydrogen ion transport in the diluting segment of frog kidney

Transepithelial H⁺ transport was studied in diluting segments of the isolated-perfused kidney ofrana esculenta. The experiments were performed in controls as well as in K⁺-adapted and Na⁺-adapted animals (exposed to 50 mmol/l KCl or NaCl, resp. for at least 3 days). Conventional and single-barreled, liquid ion-exchanger H⁺-sensitive microelectrodes were applied in the tubule lumen to evaluate transepithelial H⁺ net flux (J _te ^H ) as well as limiting transepithelial electrical and H⁺ electrochemical potential differences (PD _te ,E _te ^H ) and luminal pH at zero net flux conditions. The measurements were made in absence (control) and presence of furosemide (5·10^–5 mol/l) or amiloride (10^–3 mol/l). E _te ^H (lumen positive vs ground) was 19±3 mV in controls, 43±3 mV in K⁺ adapted but about zero in Na⁺ adapted animals. Using the correspondingPD _te -values, steady state luminal pH of 7.63±0.05, 7.13±0.05 and 8.02±0.02 was calculated for the respective groups of animals (peritubular pH 7.80). In parallel, significant secretoryJ _te ^H (from blood to lumen) was found in controls (14±2 pmol·cm^–2·S^–1) which was stimulated by K⁺ adaptation (61±8 pmol·cm^–2·s^–1) but reversed in direction by Na⁺-adaptation (–8±1 pmol·cm^–2·s^–1). Amiloride inhibited secretoryJ _te ^H . Elimination of the lumen positivePD _te by furosemide did not affect significantlyE _te ^H andJ _te ^H in control and K⁺ adapted animals but abolished reabsorptiveJ _te ^H in Na⁺ adapted animals.We conclude that in frog diluting segment H⁺ secretion is an active, amiloride-sensitive, furosemide-insensitive transport process. The data are consistent with luminal Na⁺/H⁺ exchange. The activity of this system depends critically on the metabolic state of the animal.Parts of the data were presented at the 16th Ann. Meeting of the Am. Soc. Nephrol., Washington (1983)This work was supported by österr. Forschungsrat, Proj. No.: 4366 and by Dr. Legerlotz Stiftung