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L Schild

Publications and source records attributed to L Schild.

69 records · Page 4Linked to original sources

Basolateral transport pathways for K+ and Cl- in rabbit proximal tubule: effects on cell volume.

To characterize the nature of K+ and Cl- transport pathways across basolateral membrane of rabbit proximal convoluted tubule, we used quantitative video microscopy to measure cell volume changes induced by rapid basolateral K+ and Cl- concentration changes. Elevating basolateral K+ resulted in cell swelling, which was largely inhibited by replacement of basolateral Cl- with cyclamate (85%) or by addition of 2 mM Ba2+ (72%). Substitution of basolateral Cl- by NO3- enhanced cell swelling, whereas substitution of Cl- by I- did not affect the K(+)-induced volume changes. Removal of Cl- from the bath reversed the cell swelling induced by raising K+ in the bath. Steady-state cell volume was 28% greater in hypotonic medium (250 mosmol/kgH2O) than in hypertonic medium (350 mosmol/kgH2O), and the rate of increase in cell volume induced by raising K+ was three times higher in hypotonic than in hypertonic medium. Substitution of Cl- by NO3- did not alter the effect of medium osmolality on K(+)-induced cell swelling, whereas addition of 0.2 mM diphenylamine-2-carboxylate inhibited the response (63%). We conclude that K(+)-induced cell swelling results from entry of K+ and Cl- into the cell across the basolateral membrane; it is proposed that transport of KCl across the basolateral cell membrane proceeds largely through two separate conductive pathways for K+ and Cl-. Cell swelling activates KCl transport occurring via K+ and Cl- channels across the basolateral membrane.

Animals↗

Effects of apical membrane Cl(-)-formate exchange on cell volume in rabbit proximal tubule.

We used real-time recordings of cell volume changes to test for the role of the Cl(-)-formate exchanger in mediating NaCl entry across the apical membrane of rabbit proximal tubule cells. In the absence of extracellular Cl-, 0.5 and 5 mM formate in the tubule lumen induced an increase in cell volume of 1 and 9%, respectively. Formate-induced cell swelling was reduced by alkalinizing the tubule lumen or by addition of luminal amiloride (2 mM), indicating that the increase in cell volume results from the intracellular accumulation of Na-formate via nonionic diffusion of formic acid in parallel with Na(+)-H+ exchange. The cell volume increase induced by 0.5 mM formate was potentiated (from 1 to 4%) by Cl-, as expected for a formate-mediated stimulation of NaCl uptake via parallel Cl(-)-formate exchange and Na(+)-H+ exchange across the apical membrane. By contrast, the cell volume increase induced by 5 mM formate was attenuated (from 9 to 4%) by Cl-. The attenuating effect of Cl- on formate-induced cell swelling required the operation of the apical membrane Cl(-)-formate exchanger. The effect of 1:1 Cl(-)-formate exchange to attenuate formate-induced cell swelling can be explained if the cell possesses a volume-activated anion exit pathway, most likely at the basolateral cell membrane, that is capable of mediating the efflux of Cl- but not formate from the cell.

Animals↗

Long-chain fatty acids act as protonophoric uncouplers of oxidative phosphorylation in rat liver mitochondria.

The effect of long-chain fatty acids (LCFA) on respiration and transmembrane potential (delta psi) in the resting state, and the rate of delta psi dissipation [d delta psi/dt)i) was investigated with oligomycin-inhibited rat liver mitochondria using succinate (plus rotenone) as substrate. The results obtained were compared with those of classical protonophores such as 2,4-dinitrophenol (DNP) and 4,5,6,7-tetrachloro-2-trifluoromethylbenzimidazole (TTFB). The effects of oleate or palmitate and that of DNP or TTFB on respiration and delta psi can be described by a common force-flow relationship. These facts all in all are not compatible with a decoupler-type uncoupling mechanism of LCFA; still, they indicate that the latter are protonophores. Moreover, the oleate-induced increase in the rate of delta psi dissipation closely correlates with that in respiration, suggesting that the uncoupling activity and the protonophoric activity of LCFA are interrelated. Carboxyatractyloside (CAT) exerted only a small inhibitory effect on oleate-induced respiration and delta psi dissipation, indicating that the adenine nucleotide translocase contributes to the uncoupling effect of LCFA to a minor extent only. Proton transport through the lipid region of the membrane as mediated by permeation of the protonated and deprotonated forms of LCFA is interpreted as the main process of the uncoupling of LCFA.

2,4-Dinitrophenol↗

Injury of mitochondrial respiration and membrane potential during iron/ascorbate-induced peroxidation.

First functional events during peroxidation in mitochondria consisted in a progressive inhibition of the phosphorylating and uncoupled respiration with succinate and glutamate/malate as substrates, whereas the resting state respiration during the same period was virtually not influenced. The membrane potential registered at a time with the respiration rates was capable of being built up for a relatively long time interval with only minor decreases, and broke down rather promptly when the active respiration was highly diminished. Inhibition of respiration proceeded mainly during the initiation phase of peroxidation. Lag phases of varied length, of malondialdehyde formation which were predominantly attributed to the iron/protein ratios correlated closely with different time intervals needed to attain maximal inhibition of respiration and decrease in glutathione. Hence, the lessening of respiration, drop of membrane potential and loss of the antioxidant, glutathione, represent early stages in the causal chain of events which precede the onset of intensive lipid peroxidation.

Animals↗

Kinetic limitations in the overall reaction of mitochondrial oxidative phosphorylation accounting for flux-dependent changes in the apparent delta GexP/delta mu H+ ratio.

Changes in J0, delta muH+ and delta GexP were investigated as a function of load. The flux control coefficients, particularly those of the adenine nucleotide translocator and H+-ATPase at the maximum rate of oxidative phosphorylation were seen to strongly depend on the phosphate concentration accounting in common for the highest share in flux control. There was no unique relationship observed between JP and delta muH+ in load-controlled, well coupled systems, but JP was found to depend on delta muH+ at excessive load and increasing proton leakage. All the results presented can be elucidated on the grounds of delocalized chemiosmotic coupling.

Animals↗

Chloride transport in the proximal renal tubule.

Our knowledge of chloride transport along the nephron has greatly expanded. Whereas for a long time it was assumed that chloride ions were reabsorbed entirely passively with sodium--the "mendicant" role of chloride, more recent studies suggest that several distinct reabsorptive transport mechanisms operate in parallel. Accordingly, a new model of proximal chloride transport has evolved that includes both active, transcellular as well as passive, intercellular transport pathways. Transcellular chloride reabsorption involves anion exchange mechanisms in both the luminal and peritubular cell membranes, processes that also depend on sodium, hydrogen, and bicarbonate ions. Chloride transport is thus intimately related to sodium and fluid transport as well as to cell acid-base metabolism. Unresolved problems concern the relative magnitude of transcellular and paracellular chloride transport and the details of luminal and basolateral chloride translocation steps.

Animals↗

Transport of salicylate in proximal tubule (S2 segment) isolated from rabbit kidney.

The secretory and reabsorptive transport of salicylate was studied in the isolated and perfused rabbit proximal tubule (S2 segment). Salicylate secretion (Jb----lsal) fulfilled the criteria for a carrier-mediated transport system: Jb----lsal was saturable, was reversibly inhibited by probenecid, and occurred against a concentration gradient. The Km and Vmax for this secretory transport were 80 microM and 3,200 fmol.min-1.mm-1, respectively. At luminal pH of 7.4 and 6.6, salicylate reabsorption (Jl----bsal) was low (100 fmol.min-1.mm-1). Jl----bsal was stimulated by increasing the bath PCO2 or by removing basolateral HCO3-; Jl----bsal was inhibited by ethoxyzolamide and by SITS in the bath. Our results indicate that salicylate reabsorption depends on H+ secretion, consistent with reabsorption by simple nonionic diffusion. When salicylate was present in the lumen only, Jl----bsal increased after inhibition of the secretory transport by adding ouabain or probenecid in the bath or by lowering the bath temperature. These results are compatible with luminal recycling of salicylate, and suggest the presence of a mediated secretory transporter located at the luminal membrane.

4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid↗

Effect of formate on volume reabsorption in the rabbit proximal tubule.

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.

4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid↗

Chloride transport in the mammalian proximal tubule.

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.

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Complexing of lipophilic cations and different anions as a possible cause of overestimating the transmembrane potential delta psi as measured by ion-sensitive electrodes.

The transmembrane potential delta psi of isolated rat liver mitochondria was determined by means of a dibenzyl dimethyl ammonium (DDA+)-sensitive electrode and the 86Rb+ distribution after separation by a centrifugation technique under identical incubation conditions. The delta psi determined with the electrode were higher throughout. There was no indication that the lower delta psi measured by means of 86Rb+ distribution were the result of anaerobic events connected with separation of rat liver mitochondria from the medium. The higher delta psi found by using the DDA+-sensitive electrode were due to anion-dependent changes in the activity of DDA+ ions, which were probably caused by complexing between DDA+ and different anions. A procedure was developed to correct the delta psi for this effect. The corrected values were in relatively good agreement with the delta psi obtained from the 86Rb+ distribution.

Animals↗

Secretion of tetraethylammonium by proximal tubules of rabbit kidneys.

The secretory transport of tetraethylammonium (TEA) was investigated in perfused and nonperfused isolated S1, S2, and S3 segments of proximal tubules from rabbit kidneys. In the perfused tubules the transepithelial net secretory flux and in nonperfused tubules the TEA cellular uptake were saturable (Km = 67 microM, Vmax = 2,480 fmol X min-1 X mm-1 in perfused S2 segments), energy dependent, and inhibited by mepiperphenidol. The net secretory flux of TEA (J b leads to j TEA) at a bath TEA concentration of 40 microM differed for the three segments and decreased in the order S1 greater than S2 greater than S3. The concentration of TEA in the perfusate leaving the tubule was approximately twice as great and the intracellular TEA concentration approximately 40 times as great as that in the bath. In nonperfused segments (40 microM TEA in the incubation medium) the TEA tissue water-to-medium ratio reached 100. In the three segments the ability to accumulate TEA across the peritubular membrane, thus, was similar, but the transepithelial secretory flux differed significantly. The differences in secretory rate between the three segments presumably result from differences in the luminal membrane permeability.

Animals↗