Search PubMed⌕ Search

Biomedical subjects

G Malnic

Publications and source records attributed to G Malnic.

At least 55 records · Page 3Linked to original sources
PubMed1989

Effect of amphotericin B on renal tubular acidification in the rat.

Amphotericin B, a polyene antibiotic known to induce cation-selective pore formation in biological cell membranes, was given to rats by peritoneal injection (10 mg/kg for 21-26 days) or added to luminal perfusates (2 x 10(-5) M). Kinetics of tubular acidification and alkalinization after perfusion with alkaline or acid phosphate Ringer's solution was studied by means of double barrelled antimony/reference microelectrodes in cortical distal tubules. Stationary pH increased both in early and late distal segments. Acidification and alkalinization half-times decreased markedly from 15-18 s to 6-8 s, a value similar to that found in proximal tubule. Net H-ion secretion rates as well as H-ion back-flux approximately doubled after Amphotericin B. Apparent H-ion permeability of distal tubule epithelium measured during perfusion of lumen and peritubular capillaries with phosphate Ringer's solutions doubled both in early and late segments. These data show that amphotericin B produces a distal acidification defect which impairs formation of normal transepithelial pH gradients by increasing H-ion back-flux without reducing rates of net H-ion secretion.

Amphotericin B↗
PubMed1989

Effects of adrenalectomy and acute replacement by corticosteroids on distal acidification.

The role of adrenocortical steroids in distal nephron acidification was studied in rats by measuring urine minus blood PCO2 differences (U-B PCO2) in control, sham-operated, and adrenalectomized (ADX) animals. Operations were performed 48 h before experiments. During the experiments, all rats received an infusion of 0.35-0.60 M NaHCO3, leading to urine bicarbonate concentrations in the order of 100-200 mM. Adrenalectomized rats had significantly decreased U-B PCO2 (11.9 +/- 1.99 mmHg; 1 mmHg = 133.3 Pa) with respect to sham-operated rats (39.9 +/- 1.26 mmHg). In another series, ADX rats received supplements of the adrenal steroids corticosterone, aldosterone, and 18-hydroxycorticosterone 100 min before the experiment. U-B PCO2 increased after hormone administration: corticosterone, 30.0 +/- 2.13 mmHg; aldosterone, 26.6 +/- 1.74 mmHg; 18-hydroxycorticosterone, 29.0 +/- 1.60 mmHg; but none restored these values to normal. Combinations of two hormones were also used; only aldosterone + corticosterone restored U-B PCO2 to normal: 39.0 +/- 1.66 mmHg. Renal phosphate excretion (but not urine phosphate levels) decreased significantly in ADX as compared with sham-operated rats. Extracellular volume was not significantly affected in ADX rats, which received ad libitum 0.9% NaCl for drinking. It is concluded that distal tubular acidification, as evaluated by U-B PCO2, is dependent on cortical steroids.

18-Hydroxycorticosterone↗
PubMed1989

Flow dependence of K+ secretion in cortical distal tubules of the rat.

Superficial distal tubules were pump perfused (range 0-35 nl/min), generally with solutions similar in composition to early distal tubule fluid, in control, K+-depleted, acutely K+-loaded, and K+-adapted rats with the use of double-barreled resin-reference microelectrodes to measure K+ and Na+ activities and transepithelial potential differences (PD). When perfusion rate decreased from 35 to 5 nl/min in control animals, [K+] increased from 2 mM to between 10 and 20 mM, remaining at these levels as perfusion rate was decreased further. In low-K+ rats, the change in K+ activity with flow was greatly attenuated. In K+-loaded and K+-adapted rats, [K+] was higher than in controls at all flow rates. Na+ concentrations and lumen-negative PD increased with high flow rates in control rats. Addition of 10(-3) M amiloride blocked the increase in luminal K+ with low flow rates. In the physiological range of late distal flow rates, luminal [K+] remains constant and similar to the concentration attained in the steady state. At higher flow rates, [K+] declines, and K+ balance, PD, and Na+ reabsorption modulate the relationship between K+ secretion and flow rate.

Amiloride↗
PubMed1989

Renal bicarbonate reabsorption in the rat. III. Distal tubule perfusion study of load dependence and bicarbonate permeability.

Using continuous microperfusion techniques, we studied the load dependence of bicarbonate reabsorption along cortical distal tubules of the rat kidney and their bicarbonate permeability. Net bicarbonate transport was evaluated from changes in tracer inulin concentrations and total CO2 measurements by microcalorimetry. Bicarbonate permeability was estimated from the flux of total CO2 along known electrochemical gradients into bicarbonate-and chloride-free perfusion solution containing 10(-4) M acetazolamide. Transepithelial potential differences were measured with conventional glass microelectrodes. Significant net bicarbonate reabsorption occurred at luminal bicarbonate levels from 5 to 25 mM, and at perfusion rates from 5 to 30 nl/min. Bicarbonate reabsorption increased in a load-dependent manner, both during increments in luminal bicarbonate concentration or perfusion rate, reaching saturation at a load of 250 pmol/min with a maximal reabsorption rate of approximately 75 pmol/min.mm. Rate of bicarbonate reabsorption was flow dependent at luminal concentrations of 10 but not at 25 mM. During chronic metabolic alkalosis, maximal rates of reabsorption were significantly reduced to 33 pmol/min.mm. The bicarbonate permeability was 2.32 +/- 0.13 x 10(-5) cm/s in control rats, and 2.65 +/- 0.26 x 10(-5) cm/s in volume-expanded rats. Our data indicate that at physiological bicarbonate concentrations in the distal tubule passive bicarbonate fluxes account for only 16-21% of net fluxes. At high luminal bicarbonate concentrations, passive bicarbonate reabsorption contributes moderately to net reabsorption of this anion.

Absorption↗
PubMed1988

Solvent drag of sucrose during absorption indicates paracellular water flow in the rat kidney proximal tubule.

Single convoluted proximal tubules of the rat kidney were lumen perfused in situ with isosmotic solutions containing C14-sucrose and H3-inulin as tracers, to evaluate whether the extracellular marker sucrose is entrained by water during proximal tubular reabsorption. Inulin was used as volume marker. The absorptive rate was varied by using as luminal perfusion fluids either a solution made up of (in mmole/l) 120 NaCl, 5 glucose, 25 NaHCO3 and altering the perfusion rate, or a solution containing 110 NaCl and 70 raffinose. Js, the net sucrose efflux is found to be a function of the net volume flow, Jv, such that at Jv = 0, Js is very small and at high rates of Jv, Js is over 60-fold the value observed at low Jv values. In addition, the transported to luminal sucrose concentrations decreased with Jv in a hyperbolic manner. Unstirred layers affect the diffusive component of Js, but only to a small extent. Therefore, the large remaining dependency of Js with Jv must be due to drag of sucrose by water, within the paracellular pathway. This leads to the conclusion that water flows through the paracellular pathway during absorption in the rat proximal tubule, in addition to transcellular water flow. Using equations for molecular sieving and the measured value of sigma s for sucrose of 0.76-0.91, it is calculated that the pathway where entrainment of solute by water occurs must be 1.0-1.1 nm wide. This calculation is only tentative since sigma s depends on the as yet unknown relative contribution of transcellular and paracellular pathways to transepithelial water osmotic permeability.

Absorption↗
PubMed1988

Sodium dependence of early distal H+ secretion in rat kidney.

In order to study the mechanism of H-ion secretion in cortical distal tubules of the rat kidney, the luminal pH and transepithelial potential difference (PD) were measured with double-barrelled, pH-sensitive, resin/reference microelectrodes. Perfusion of peritubular capillaries with low-sodium solutions increased luminal pH by 0.28 +/- 0.024 units. Perfusion of the lumen with 1 mM amiloride increased luminal pH by 0.67 +/- 0.01 units. These changes could not be attributed to modification of transepithelial PD. We conclude that early distal acidification is sodium-dependent, possibly owing to the presence of Na+/H+ exchange.

Animals↗
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↗
PubMed1987

H+ secretion in distal tubules: the effect of carbonic anhydrase in luminal perfusates.

1. The ability of distal tubules in control rats to acidify luminal fluids is questioned in view of the absence of bicarbonate reabsorption and of a disequilibrium pH as shown by the equilibrium electrode method. However, the ability to establish a transepithelial pH gradient has been shown previously. These discrepancies are investigated in the present work. 2. Luminal pH was measured in cortical distal segments of rat nephron during stationary perfusion with carbonic anhydrase (1 mg/ml)-containing solutions. During perfusion with bicarbonate Ringer, the stationary pH was significantly higher in the presence of the enzyme, suggesting the existence of an acid disequilibrium pH in these segments. 3. Acidification half-times were also longer in the presence of the enzyme, yielding bicarbonate reabsorption rates of 0.24 +/- 0.07 (N = 29)nmol cm-2s-1 in early and 0.32 +/- 0.09 (N = 28) in late distal tubules. 4. During luminal perfusion with phosphate Ringer, significant transepithelial pH gradients were also observed but were unaffected by the enzyme. Thus, a disequilibrium pH was detected in distal tubules when perfused with bicarbonate-containing solutions. This acid disequilibrium pH can inhibit distal bicarbonate reabsorption by creating an adverse pH gradient. 5. In conclusion, the ability of distal tubules to establish a significant pH gradient will contribute to the titration of non bicarbonate buffers, i.e., to titratable acid formation.

Animals↗
PubMed1987

Effect of corticosteroids on proximal tubular acidification in the rat.

The role of corticosteroid hormones in proximal tubular acidification was studied in control, sham-operated and adrenalectomized (ADX) rats by means of the stopped-flow microperfusion technique and by measurement of tubular acidification kinetics with pH microelectrodes. Rats were adrenalectomized 48 h before experiments, and aldosterone, corticosterone and 18-OH corticosterone (18-OH-B) were given intramuscularly 100 and 40 min before starting the experiments. Stationary proximal luminal pH increased from 6.78 to 7.03 after ADX, while acidification half-times (t1/2) increased from 4.41 to 11.43 s. In consequence, net bicarbonate reabsorption fell from 2.18 to 0.50 nmol/cm2 X s after ADX. The administration of the three hormones stimulated tubular acidification, bicarbonate reabsorption reaching 1.18-1.28 nmol/cm2 X s. These data suggest that proximal tubular acidification is stimulated by both mineralo- and glucocorticoid hormones and by 18-OH-B; their mechanisms of action is discussed on the basis of a recently described model.

11-Hydroxycorticosteroids↗
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↗
PubMed1985

Transfer of base across the basolateral membrane of cortical tubules of rat kidney.

The transfer of HCO3-/OH- across the peritubular membrane of rat cortical tubules was studied by measuring capillary pH during stopped-flow microperfusion of peritubular capillaries with electrolyte solutions containing 3 mM HCO3-. The rate of alkalinization of these solutions was significantly delayed when 10(-4) M acetazolamide, 5 X 10(-4) M SITS or 2 mM Ba2+ were added, as well as when chloride was substituted by gluconate. Under these conditions, stationary capillary pH was slightly but significantly increased. In another series, the lumen of proximal tubules was perfused with alkaline solutions while pH was measured in adjacent capillaries. During perfusion, capillary pH rose to a level 0.4 units higher than in free-flow conditions, returning after filling the lumen with oil; the rate of capillary pH return to baseline is a measure of base extrusion from cells, in the absence of influx from the lumen. This rate is also significantly delayed by acetazolamide. The data show that peritubular base extrusion is dependent on carbonic anhydrase, on basolateral membrane voltage, and on interstitial chloride, and delayed by the anion exchange inhibitor SITS; they are compatible with both basolateral HCO3-/Cl- exchange and conductive bicarbonate transfer.

Animals↗
PubMed1985

Dinitrophenol effect on proximal tubular acidification in the rat.

The effect of 10(-3) M-dinitrophenol (DNP) on renal tubular acidification was studied in proximal tubules of rat kidneys perfused with mammalian Ringer solution. Alkaline (pH 7.8) or acid (pH 5.8) phosphate-buffered solutions were injected into the lumen, and pH changes recorded with antimony micro-electrodes. Luminal perfusion with DNP caused complex acidification or alkalinization curves, an initial rapid shift toward a higher than control pH being followed by a slower acidification. Acidification half-times of the initial phase (t1/2 = 1.6 s) were markedly shorter than controls (6.2 s). This response was probably due to transient action of DNP, since keeping constant peritubular DNP levels by capillary perfusion caused simple exponential pH curves. In such experiments luminal pH increased from pH 6.6-6.8 to 7.1-7.2, while acidification and alkalinization t1/2 decreased from about 7 s to 3-5 s. Secretory H-ion fluxes increased transiently and then fell below controls after a few minutes of perfusion, while H-ion efflux from the lumen increased progressively. These data suggest that, besides its known effect on cell metabolism, DNP acts directly on proximal tubular cell membranes, increasing the rate of passive H-ion equilibration, both mechanisms impairing the tubular capacity to maintain normal proximal pH gradients and fluxes.

2,4-Dinitrophenol↗
PubMed1985

Verapamil effect on renal function of normotensive and hypertensive rats.

The effect of verapamil, a Ca++ antagonist drug, on renal function and proximal fluid reabsorption in normal and hypertensive (GII) rats was studied. During intravenous infusion of verapamil, mean arterial pressure (MAP) fell significantly in both groups, 23% more in hypertensive than in normotensive rats. Glomerular filtration rate (GFR) was significantly higher in hypertensive rats and also increased significantly in this group during verapamil infusion. Effective renal plasma flow (ERPF) was similar in both groups and did not change significantly during verapamil infusion. The increase in urine flow, Na+ and Ca++ excretion was higher in hypertensive than in normotensive rats during verapamil infusion. When 10(-5) M verapamil was added to the luminal perfusate of proximal tubules, fluid reabsorption was reduced to 64% in normotensive and to 42% in hypertensive rats. When added to capillary perfusate, fluid reabsorption was almost completely but reversibly inhibited (92% in normotensive and 83% in hypertensive rats). Our findings indicate a direct effect of verapamil on renal Na+ and possibly also on Ca++ reabsorption, suggesting involvement of the Na+-Ca++ countertransport system. The greater effect of verapamil on Na+ excretion in hypertensive rats was not due to increased action on proximal Na+ reabsorption.

Absorption↗
PubMed1984

Proximal tubular HCO3-, H+ and fluid transport during maleate-induced acidification defect.

The mechanism of tubular acidification was studied in proximal tubular acidification defect induced in rats by acute parenteral infusion of maleate (200 mg/kg), which causes diuresis and bicarbonaturia. Proximal tubular bicarbonate reabsorption and H+ ion secretion were determined by stopped-flow microperfusion and measurement of luminal pH by Sb microelectrodes. Stationary pH increased in proximal tubule from 6.78 to 7.25 and bicarbonate reabsorption decreased from 1.32 to 0.51 nmol/cm2 X s. In these segments, mean cell PD fell from -66.6 to -20.2 mV, while Jv as estimated by the Gertz technique fell to 15% of controls. A similar impairment of acidification was observed during luminal and capillary perfusion with phosphate Ringer's. Since H+-ion efflux from the lumen was not significantly increased and both acidification and alkalinization half-times (t/2) were increased, no evidence for an increase in passive permeability for H+/HCO3- was obtained. The increased t/2 found during luminal perfusion with acid phosphate indicates, according to an electrical analog model, a reduction in pump series conductance. These results show that maleate affects both proximal Na+ and H+ transport; this effect may be ascribed to impairment of sodium-dependent transport systems in the brush-border membrane.

Acid-Base Equilibrium↗
PubMed1984

Factors affecting proximal tubular acidification of non-bicarbonate buffer in the rat.

The effect of peritubular PCO2 and pH changes within the physiological range on proximal tubular acidification of non-bicarbonate (phosphate) buffer was evaluated with and without carbonic anhydrase inhibition by stopped-flow microperfusion and Sb micro-electrode techniques. Luminal steady-state pH was reduced from 6.69 to 6.37 and H ion fluxes (JH+) increased from 0.63 to 1.57 nmol cm-2 s-1 by increasing capillary CO2 from 0 to 9.6% at pH 7.2. After acetazolamide a marked, although attenuated, effect of CO2 on acidification was still observed; JH+ increased from 0.088 nmol cm-2 s-1 at 0% CO2 to 0.78 at 9.6% CO2. Most of this effect can be explained by titration of luminal buffer by CO2, uncatalysed CO2 hydration and H2CO3 recirculation. An increase in capillary CO2 reduced acidification half-times (t/2), which, according to an analogue circuit model, may be due to increased H ion access to the pump. Peritubular pH changes at 0% CO2 also modified tubular acidification, increasing JH+ from 0.73 nmol cm-2 s-1 at pH 7.6 to 0.99 at pH 7.0. After acetazolamide, JH+ still increased from 0.11 nmol cm-2 s-1 at pH 7.6 to 0.57 at pH 7.0. In conclusion, both peritubular CO2 changes at constant pH and pH changes at 0% CO2 were effective to modify JH+, in the presence and absence of carbonic anhydrase activity. In the studied range, capillary CO2 induced larger changes in JH+ than pH. The data show substrate (H ion) is a limiting factor for tubular H ion secretion.

Acetazolamide↗