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G Giebisch

Publications and source records attributed to G Giebisch.

At least 163 records · Page 9Linked to original sources
PubMed1987

Na-dependent effects of DOCA on cellular transport properties of CCDs from ADX rabbits.

Microelectrode methods were used to assess the Na-dependent effects of mineralocorticoids on the conductive transport properties of cortical collecting ducts (CCD) from adrenalectomized (ADX) rabbits. Rabbits were divided into four groups: control, 3 h treatment with deoxycorticosterone (DOCA), 18 h DOCA treatment, and 18 h DOCA plus amiloride treatment. After only 3 h of DOCA treatment the transepithelial potential (VT) increased significantly from -1.4 to -8.7 mV, and after 18 h VT increased further to -19.3 mV. The basolateral membrane potential (Vb) significantly increased after 18 h (from -61.6 to -87.6 mV). However, compared with treatment with DOCA alone for 18 h, both -VT and -Vb were significantly attenuated with DOCA plus amiloride treatment to -8.3 and -72.9 mV, respectively. The apical membrane Na and K currents (INaa and IKa) and conductances (GNaa and GKa) were all increased after 3 h DOCA treatment. After 18 h of DOCA treatment, IKa, INaa, and GKa increased further, but GNaa remained unchanged. On simultaneous treatment with DOCA and amiloride, the maximum pump current (Ipmax), IKa, INaa, and GKa but not GNaa were all reduced significantly. These results support a mechanism whereby mineralocorticoids first increase GNaa. A rise in cell Na results then in the simultaneous increase in GKa and sodium transport, probably by insertion of new Na pumps. An additional direct effect of DOCA on GKa and Ipmax is not excluded.

Adrenalectomy↗
PubMed1987

The collecting tubule of Amphiuma. I. Electrophysiological characterization.

Single collecting tubules of Amphiuma kidneys were perfused in vitro to characterize their electrophysiological properties. The lumen-negative potential (-24 mV) was abolished by amiloride in the lumen and by ouabain in the bath. Ion substitution experiments in the lumen demonstrated the presence of a large sodium conductance in the apical cell membrane, but no evidence was obtained for a significant potassium or chloride conductance. Ion substitutions in the bath solution and the depolarizing effect of barium on the basolateral membrane potential demonstrated the presence of a large potassium conductance in the basolateral cell membrane. Measurements of dilution potentials in amiloride-treated tubules revealed a modest cation selectivity of the paracellular pathway. These results support a cell model in which sodium reabsorption occurs by electrodiffusion across the apical cell membrane and active transport across the basolateral cell membrane. The absence of a detectable potassium conductance in the apical cell membrane suggests that secretion of this ion cannot take place by diffusion from cell to lumen.

Algorithms↗
PubMed1987

The collecting tubule of Amphiuma. II. Effects of potassium adaptation.

Electrophysiological and transport properties were studied in isolated and perfused Amphiuma collecting tubules from two groups of animals, one exposed to a high sodium (NA), the other to a high potassium (KA) environment (both conditions known to modulate blood aldosterone levels). The transepithelial lumen-negative potential was significantly larger (-38 +/- 5 mV) in tubules from KA animals than from NA animals (-15 +/- 3 mV). In addition, we observed an increase in the apical amiloride-sensitive sodium conductance and stimulation of the transepithelial sodium current. Although no measurable potassium conductance was found in the apical cell membrane in either group, a potassium selectivity of the paracellular transport pathway was observed in the KA animals. Net potassium secretion was demonstrated in KA tubules (helium-glow photometry and [3H]inulin analysis). Potassium secretion was abolished by luminal amiloride but imposition of a bath-to-lumen potassium gradient induced potassium secretion. We conclude that in contrast to the mammalian cortical collecting tubule in which potassium secretion is largely transcellular, potassium secretion in the Amphiuma collecting tubule is by diffusion through the paracellular pathway.

Adaptation, Physiological↗
PubMed1987

Potassium-sparing diuretics.

Amiloride, triamterene, and the spirolactones are potassium-sparing diuretics which act on the distal parts of the nephron, from the late distal tubule to the collecting duct. In these segments, active sodium reabsorption occurs through the following mechanism: sodium ions enter the cell through specific channels present in the luminal membrane and are extruded out of the cell into the peritubular medium by a sodium-potassium exchange pump, the Na-K-ATPase. Amiloride in micromolar concentrations reduces the sodium transport by blocking the luminal membrane sodium channel. Triamterene has a similar effect, although with a lower affinity; the available studies do not allow to determine if an inhibitory effect of triamterene on the Na-K-ATPase plays an additional role in its diuretic action. The spirolactones are competitive inhibitors of aldosterone, the mineralocorticoid hormone which promotes sodium reabsorption by increasing both the number of active sodium channels in the luminal membrane and the number of active Na-K pumps in the peritubular membrane. By the inhibitory effect on the electrogenic sodium transport, amiloride, triamterene, and the spirolactones decrease the lumen-negative transepithelial potential difference. This reduces the driving force for potassium movement into the tubular lumen and thus decreases potassium excretion.

Aldosterone↗
PubMed1987

Independent effects of aldosterone and potassium on induction of potassium adaptation in rat kidney.

We examined the independent effects of a high potassium diet and increased aldosterone levels on the development of renal potassium adaptation. This condition is defined by the increased ability of the kidneys to excrete an acute infusion of potassium. Rats were adrenalectomized (ADX) and received aldosterone at basal levels (0.5 microgram/100 g X d) or at high levels (2.0 micrograms/100 g X d) for 10 d. In each experimental group, animals received either a control diet or a high potassium diet. In ADX animals with basal aldosterone levels, a high potassium intake increased but did not completely restore the ability to excrete potassium and induced proliferation of the basolateral membrane of principal cells in the collecting tubule (i.e., morphologic adaptation). In contrast, increased aldosterone did not induce functional adaptation. Elevated aldosterone and dietary potassium intake were required to produce functional potassium adaptation indistinguishable from that in potassium-loaded, adrenal-intact animals.

Adaptation, Physiological↗
PubMed1987

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↗
PubMed1987

Renal bicarbonate reabsorption in the rat. II. Distal tubule load dependence and effect of hypokalemia.

We studied two groups of rats acutely loaded with bicarbonate, control rats on a standard diet and rats kept on a K-free diet for 3 wk. Compared with controls, K-depleted rats had reduced fractional excretion of bicarbonate despite their elevated filtered bicarbonate load. Distal bicarbonate reabsorption increased in K-depleted rats. In the presence of almost identical early distal bicarbonate loads (481 +/- 40 pmol/min in controls and 444 +/- 50 pmol/min in K depletion), distal bicarbonate reabsorption was significantly enhanced in K depletion (247 +/- 17 pmol/min) as compared with controls (179 +/- 18 pmol/min). These values are significantly different from each other, and both are severalfold higher than bicarbonate reabsorption in nonloaded conditions. In conclusion, distal bicarbonate reabsorption is load dependent, and distal bicarbonate reabsorption is stimulated in K depletion.

Animals↗
PubMed1987

Filtered load of buffer and renal H-ion secretion: mechanism of proximal tubule load dependence.

When the filtered load of buffers like bicarbonate or phosphate is increased by elevating GFR or buffer concentration in plasma, the overall renal reabsorption of bicarbonate or the formation of titratable acidity are markedly increased. The same happens when buffer concentration or flow rate are varied during proximal microperfusion. We have recently studied the mechanisms of this functional dependence. We have observed that the rate of bicarbonate reabsorption is always proportional to luminal buffer concentration when a stationary fluid column is injected into the proximal lumen. H-ion secretion is also proportional to luminal levels of non-bicarbonate buffers. Using a pH-stat technique adapted to renal tubules, we have shown that H-ion secretion is dependent on proximal pH independently of the used buffer species. A kinetic analysis of these data shows a non-linear relationship between luminal H+ and H+ secretion, compatible with carrier mediated transport.

Animals↗
PubMed1986

Mechanisms of renal tubular acidification.

Both bicarbonate retrieval from the filtrate as well as the net excretion of acid depend upon hydrogen ion secretion by the tubular epithelium. Hydrogen ion secretion is mediated either by sodium-hydrogen exchange, an electroneutral and secondary active process, or by hydrogen ion secretion, a directly electrogenic and primary active process. Extrusion of hydrogen ions across the apical cell membrane is accompanied by electrogenic bicarbonate transfer across the basolateral cell membrane. Both luminal and peritubular pH exert a strong influence upon acidification by altering the gradient against which hydrogen transport or base exit occur. In the distal nephron, both hydrogen ion secretion and bicarbonate secretion may occur. These transport operations have been shown to be mediated by subgroups of intercalated cells in which hydrogen pumps and bicarbonate-chloride exchange processes are located either in the apical or basolateral cell membranes. Regulation of acidification involves several factors: the rate of luminal buffer delivery, sodium and chloride delivery, the luminal and peritubular pH and pCO2, the electrical potential, mineralocorticoids and the state of the potassium balance.

Acid-Base Equilibrium↗
PubMed1986

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.

Animals↗
PubMed1986

Regulation of single potassium ion channels from apical membrane of rabbit collecting tubule.

The regulation of K+-channel activity from the apical membrane of the rabbit cortical collecting tubule was studied using the patch-clamp technique. Using inside-out patches, channel open probability was determined as a function of calcium and barium concentration and transmembrane potential. Channel open probability was increased by raising bath (cytoplasmic) calcium concentration, with an apparent Ka of 2.4 microM. Mean channel open time also increased during this maneuver. The channel was reversibly inhibited by barium, applied to the cytoplasmic face, with an apparent Ki of 12 microM. Depolarization of the transmembrane potential increased channel open probability. With 1 mM calcium in the bath solution, the open probability was one-half maximal at -55 mV. It is concluded that this channel is the probable route for transcellular K+ secretion by the cortical collecting tubule and that procedures likely to increase intracellular calcium and/or depolarize the apical membrane will cause an increased potassium secretion.

Animals↗
PubMed1986

Electrophysiological heterogeneity of proximal convoluted tubules in Amphiuma kidney.

The present study was designed to identify functional differences between dark (early to mid) and white (late) proximal tubule segments in Amphiuma kidney. The potential difference across the peritubular cell membrane (Vb), the luminal cell membrane (Va), and the epithelium (Vte) are not significantly different between dark and white segments. Cellular and luminal cable analysis reveals that the resistance of the cell membranes in parallel is lower in dark (28.6 +/- 3.2 k omega X cm) than in white segments (63.2 +/- 5.0 k omega X cm) in contrast to the transepithelial resistance, which is higher in dark (26.6 +/- 5.5 k omega X cm) than in white (3.5 +/- 0.7 k omega X cm) segments. A step-increase of peritubular potassium (from 2.5 +/- 12.5 mmol/liter) depolarizes Vb more in white (20.1 +/- 1.2 mV) than in dark (7.2 +/- 0.4 mV) segments, whereas addition of bicarbonate to peritubular perfusate hyperpolarizes Vb more in dark (-22.4 +/- 1.6 mV) than in white (-5.9 +/- 0.7 mV) segments. An increase of luminal potassium depolarizes Va more in dark (21.3 +/- 2.0 mV) than in white (9.3 +/- 1.9 mV) segments. Similarly luminal glucose depolarizes Va more in dark (10.7 +/- 1.2 mV) than in white segments (3.2 +/- 1.4 mV). Partial peritubular replacement of NaCl and reduction of peritubular chloride polarize Vte more in white (9.6 +/- 1.0 and 28.9 +/- 2.9 mV) than in dark segments (7.0 +/- 0.5 and 15.5 +/- 1.9 mV). In conclusion, compared with white segments, dark segments have lower cell membrane and higher shunt resistances, lower potassium and higher bicarbonate conductances of the peritubular cell membrane, and a higher capacity to reabsorb glucose. Paracellular shunt chloride conductance is relatively high in both segments.

Animals↗
PubMed1986

Renal bicarbonate reabsorption in the rat. I. Effects of hypokalemia and carbonic anhydrase.

Free-flow micropuncture studies were carried out on superficial rat proximal and distal tubules to assess the participation of different nephron segments in bicarbonate transport. Particular emphasis was placed on the role of the distal tubule, and micro-calorimetric methods used to quantitate bicarbonate reabsorption. Experiments were carried out in control conditions, during dietary potassium withdrawal, and after acute intravenous infusions of carbonic anhydrase. We observed highly significant net bicarbonate reabsorption in normal acid-base conditions as evidenced by the maintenance of significant bicarbonate concentration gradients in the presence of vigorous fluid absorption. Distal bicarbonate reabsorption persisted in hypokalemic alkalosis and even steeper transepithelial concentration gradients of bicarbonate were maintained. Enhancement of net bicarbonate reabsorption followed the acute intravenous administration of carbonic anhydrase but was limited to the nephron segments between the late proximal and early distal tubule. The latter observation is consistent with a disequilibrium pH along the proximal straight tubule (S3 segment), the thick ascending limb of Henle, and/or the early distal tubule.

Absorption↗
PubMed1986

Potassium channels along the nephron.

The K+ channels that are present in three different nephron segments, the Necturus proximal, Amphiuma early distal (diluting segment), and rabbit collecting tubule have been examined. Ca2+-sensitive K+ channels were present in the apical membranes of the cells lining all these segments. The channels were all voltage-sensitive and their open probability increased with membrane depolarization. Because of the ubiquitous distribution, it is suggested that this channel is responsible for K+ secretion by the nephron and that the same intracellular regulators act throughout the various segments. Basolateral K+ channels have been examined only in Necturus proximal tubules. This channel is apparently insensitive to Ca2+; the voltage dependence is exactly opposite to that of the apical K+ channels; that is, hyperpolarizing potentials caused an increase in open probability. These differences in regulatory factors permit the independent regulation of apical and basolateral membrane K+ permeabilities that must occur in renal cells.

Animals↗
PubMed1985

Hormonal control of distal nephron function.

The distal tubule and collecting tubules are important control sites of fluid and electrolyte excretion. In our presentation we consider the cell mechanisms of transport of sodium and potassium ions and the effects of several hormones. Aldosterone and antidiuretics stimulate potassium secretion directly, and the available evidence strongly suggests that this effect involves the principal cell population. Epinephrine inhibits potassium secretion at sites beyond the distal tubule. In addition to such direct effects, secondary factors such as hormone-induced changes in flow rate along the distal tubule and changes in the plasma potassium level play an important modifying role. Several examples are presented to demonstrate that interaction of several control components uncouples potassium secretion from distal flow rate and tends to stabilize urinary potassium excretion during changes in sodium and water balance.

Acid-Base Equilibrium↗
PubMed1985

Cellular electrophysiology of potassium transport in the mammalian cortical collecting tubule.

Electrophysiological studies were carried out on single perfused cortical and medullary collecting ducts to define the potassium and sodium transport properties of their apical and basolateral cell membranes. In addition, the effects of chronic mineralocorticoid hormone treatment on the mechanism of transport of potassium ions were evaluated. Studies included the measurement of transepithelial and cell potentials, and the resistance of individual cell membranes. The apical cell membrane of principal cells of the cortical collecting duct is characterized by separate potassium and sodium conductances. The basolateral cell membrane has also a potassium conductance, whereas the intercellular shunt pathway is largely permeable to chloride ions. Stimulation of potassium secretion by mineralocorticoids is associated with the following events. Increased cell potassium uptake across the basolateral cell membrane due to stimulation of Na-K ATPase and a more favorable electrical driving force for passive entry, facilitated exit of potassium from the cell to the tubule lumen by a more favorable electrochemical gradient (apical cell membrane depolarization) and enhanced potassium secretion by in increase of the potassium conductance of the apical cell membrane. Some properties of single potassium channels in the apical membrane of rabbit cortical collecting tubules are also described.

Animals↗