Biomedical subjects

W Van Driessche

Publications and source records attributed to W Van Driessche.

At least 73 records · Page 4Linked to original sources

PubMed1991

Cyclic AMP increases electrical capacitance of apical membrane of toad urinary bladder.

We have measured the effects of oxytocin and three other compounds (chlorophenyl-thio-cyclic AMP, forskolin and theophylline) that increase cytoplasmic cyclic AMP on the impedance of the toad urinary bladder. Membrane capacitance was calculated from transepithelial impedance measured by a computerized sine wave method. All four agents increased tissue capacitance. Since in these tissues this parameter is proportional to apical membrane area our results suggest that cAMP can be a second messenger involved in the action of agents that promote fusion of exocytotic vesicles with the apical membrane.

Animals↗

PubMed1990

Ca2+ entry through the apical membrane reduces antidiuretic hormone-induced hydroosmotic response in toad urinary bladder.

The role of Ca2+ in the regulation of antidiuretic hormone(ADH)-induced water permeability of the apical membrane of the toad urinary bladder was examined. The effects of modifying Ca2+ entry through the apical membrane of toad urinary bladders on the hydroosmotic water flow (phi H2O) and short circuit current (Isc) were measured. In most experiments the bladders were treated with small amounts of Ag+ (10(-7) mol/l) on the apical side. This treatment was used because previous experiments indicate that it markedly increases alkali-earth cation fluxes through an amiloride-insensitive cation channel in the apical membrane of the urinary bladder. Moreover, when Ca2+ is the major cation in the apical solution of these Ag(+)-treated bladders, Isc is mostly due to Ca2+ entry through the apical membrane. Ag+ increased Isc and simultaneously inhibited phi H2O in bladders perfused with Ca2+ solutions on the apical side. Addition of La3+ to the apical solution reversed the stimulation of Isc and the inhibition of phi H2O produced by Ag+. When bladders were perfused with Ca2(+)-free solutions on the apical side, addition of Ag+ did not inhibit phi H2O while the stimulation of cation movements through the amiloride-insensitive cation channel persisted. In bladders perfused with apical Ca2+ solutions and treated with chlorophenyl thio-cyclic adenosine monophosphate (ClPheS-cAMP) the addition of Ag+ did not inhibit phi H2O while it still increased Isc. Finally, addition of Ca2+ to the apical solution of bladders not treated with Ag+ reduced phi H2O.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1990

Effects of millimolar concentrations of glutaraldehyde on the electrical properties of frog skin.

1. The effects of millimolar concentrations of glutaraldehyde on the electrophysiological properties of the epithelium of frog skin (Rana temporaria) were investigated. We recorded short-circuit current (Isc), transepithelial conductance (Gt) and impedance (Zt), fractional resistance (fRo) and the potential difference across the apical membrane (Vo). We used either Na+ or K+ as major mucosal cations to compare the effects on transepithelial Na+ and K+ currents (INa and IK) and thus on the apical Na+ and K+ permeabilities. 2. At concentrations above 0.005% (w/v) or 0.5 mM, glutaraldehyde irreversibly and completely inhibits both INa and IK within 2-3 h. The initial time courses of the inhibition of transepithelial currents following serosal and mucosal applications of the compound markedly differ. 3. Glutaraldehyde decreased Gt in sulphate Ringer solutions while it augmented Gt severalfold in chloride Ringer solution. 4. Measurements of the transepithelial impedance of tissues incubated with sulphate solutions showed that glutaraldehyde increased the resistances of both apical and basolateral membranes significantly. The capacitance of the apical membrane was augmented, while the basolateral membrane capacitance was drastically decreased. 5. Microelectrode impalements of the granulosum cells showed that glutaraldehyde decreased Vo by more than 40 mV and increased fRo, which reached values around 90%. 6. The role of free amino groups in ion-transporting proteins and the potential non-fixative uses of protein cross-linkers in epithelia are discussed.

Animals↗

PubMed1989

Intracellular potentials of toad urinary bladder.

Short-circuited urinary bladders of Bufo marinus were impaled with microelectrodes. Intracellular potentials in the order of -80 mV were recorded. On inhibition of apical Na entry they hyperpolarized by about 15 mV and the fractional resistance of the apical membrane increased to near 1.0. These patterns are similar to those of other tight Na transporting amphibian epithelia. On two occasions, stable recordings from a single cell were obtained before and after oxytocin. Intracellular potential and fractional resistance of the apical membrane remained constant despite doubling of Na transport and cellular conductance. This finding suggests that oxytocin stimulates conductive pathways in both, apical and basolateral cell membranes of the toad urinary bladder.

Action Potentials↗

PubMed1989

Effect of amiloride on the poorly selective cation channel of larval bullfrog skin.

A small, inward-directed, short-circuit current (SCC) was measured across the isolated skin of larval bullfrogs (Rana catesbeiana) when either NaCl or KCl Ringer solution bathed the mucosal surface. The addition of amiloride, in concentrations of 1-100 microM, produced a stepwise increase in SCC. As SCC values became maximally elevated by amiloride, the plateau value (So) of the Lorentzian component in the power-density spectrum increased, whereas the corner frequency (fc) decreased. This agonist effect of amiloride can be explained by an increase in the open probability and possibly the single-channel current of the larval channel. When the amiloride concentration was increased above 100 microM, the SCC values declined progressively but usually remained above pretreatment values. This suggests an antagonist effect of amiloride that is concurrent with the agonist effect. The removal of Ca2+ from the mucosal Ringers increased SCC in conjunction with an increase in So and a decrease in fc. Under these conditions, the maximal agonist effect of amiloride was observed at concentrations of 10-20 microM. Ca2+ thus exerts an inhibitory effect on the larval cation channel that interferes with the agonist effect of amiloride. The addition of Ba2+ to Ca2+-free preparations lowered SCC and reduced the agonist effect of amiloride.

Amiloride↗

PubMed1989

Evidence for apical sodium channels in frog lung epithelial cells.

To reveal the mechanism of Na+ transport across Xenopus lung epithelium, we recorded short-circuit current (Isc), transepithelial resistance (Rt), and current noise spectra while the isolated lung tissues were mounted in an Ussing-type chamber. Mean values of Isc and Rt obtained while the tissue was bilaterally incubated with NaCl-Ringer solution were Isc = 11.57 +/- 1.19 microA.cm-2 and Rt = 0.82 +/- 0.07 k omega.cm2. Amiloride added to the mucosal (apical) side depressed Isc by 61 to 99%. Ouabain abolished Isc totally when added to the basolateral compartment. Adenosine 3',5'-cyclic monophosphate (cAMP), epinephrine, and a variety of other compounds did not alter Isc significantly. Transepithelial depolarization with serosal KCl solution reduced Isc to 6.22 +/- 1.37 microA.cm-2. Amiloride-sensitive current and the kinetics of amiloride interaction were not significantly affected by depolarization. Fluctuation analysis of Isc in the presence of amiloride revealed a Lorentzian component in the power density spectrum indicating apical Na+ channels. Assuming pseudo-first order kinetics, we calculated single channel currents (iNa) and channel density (M): iNa = 0.29 +/- 0.04 pA and M = 0.24 +/- 0.04 micron 2. Our results show that the route for Na+ transport through lung epithelial cells follows the classical Koefoed-Johnson-Ussing model for tight epithelia.

Amiloride↗

PubMed1989

Forskolin activates gated Cl- channels in frog skin.

We investigated the effect of forskolin on Cl- movements across the isolated epithelium of frog skin. With Cl- on both sides, forskolin (50 mumol/l) increased the transepithelial conductance considerably and elicited significant Cl- secretion. Establishing transepithelial Cl- gradients markedly increased the Cl- currents (ICl). During forskolin treatment, the power density spectra (PDS) of the fluctuation in transepithelial current contained a Lorentzian component that depended on the presence of Cl- in the bathing solutions. Mucosal as well as serosal diphenylamine-2-carboxylic acid (DPC; 1 mmol/l) partially depressed ICl as well as the Lorentzian noise component. Microelectrode recordings from cells involved in transepithelial Na+ absorption showed that forskolin activates gated Cl- channels in a cellular pathway in parallel with the Na+-transporting granulosum cells of the frog skin. The activation of the Cl- -dependent currents and Lorentzian noise was rather variable, and adaptation of the animals to solutions that contained 40 or 60 mmol/l NaCl increased the sensitivity to forskolin. In skins of salt-adapted animals, oxytocin (0.1 U/ml) also slightly activated the Cl- pathway. On the other hand, oxytocin and 8-(4-chlorophenylthio)adenosine 3',5'-cyclic monophosphate (CPT-cAMP; 1 mmol/l) were without effect in control skins.

Animals↗

PubMed1989

Exocytotic events unrelated to regulation of water permeability in amphibian tight epithelia: effects of oxytocin, PMA and insulin on membrane capacitance, water and Na+ transport.

We measured the effects of oxytocin on capacitance and hydroosmotic water flow in the urinary bladder of the toad Bufo marinus and the skins of Rana pipiens and Rana temporaria. Oxytocin increased capacitance in all these tissues but stimulated hydroosmotic water flow only in the urinary bladder. We also measured the effects of oxytocin and PMA on the capacitance and hydroosmotic water flow of the toad urinary bladder. Both agents produced increases in membrane capacitance that were additive, however, PMA produced a stimulation of water flow that was only a fraction of that caused by oxytocin. Comparison of the effects of PMA and insulin in toad urinary bladder showed that in contrast with PMA, insulin did not increase membrane capacitance in this tissue. Moreover, insulin stimulated Isc in the urinary bladder while PMA produced an inhibition of variable magnitude. These results suggest that: (1) oxytocin can promote the fusion with the apical membrane of cytoplasmic membranes with or without water channels; (2) oxytocin and PMA stimulate the fusion with the apical membrane of cytoplasmic membranes originating in different pools; membranes in each pool have different water permeabilities and their insertion is controlled by different signals; (3) PMA and insulin act through different mechanisms in the toad urinary bladder.

Animals↗

PubMed1988

Circadian rhythm of apical Na-channels and Na-transport in rabbit distal colon.

In vivo and in vitro studies showed that electrogenic sodium transport in rabbit distal colon is modulated by aldosterone. It varies in a circadian rhythm; the external synchronizer is the light-dark cycle. The site of regulation was found to be in the apical membrane of colonic epithelial cells, in which the number of conducting sodium-channels is increased by aldosterone.

Aldosterone↗

PubMed1988

Current-voltage relations of Cs+-inhibited K+ currents through the apical membrane of frog skin.

The voltage-dependence of the inhibitory effect of mucosal Cs+ on the inward K+ current through the apical membrane of frog skin (Rana temporaria) was studied by recording transepithelial current-voltage relations. Experiments were performed with skins exposed to NaCl and KCl Ringer solutions on the serosal and mucosal side respectively (control skins), as well as with tissues incubated with K2SO4 Ringer solutions on both sides (depolarized skins). Studies of the dose-dependence of the Cs+ block showed that under both experimental conditions the apparent affinity of Cs+ increased as the transepithelial potential was clamped at higher mucosal positive voltages. Under control conditions, the concentration of Cs+ required to block 50% of the K+ current (KCs) recorded while the transepithelial voltage was clamped at zero mV was 16 mmol/l. KCs decreased exponentially with mucosal positive voltages. The dependence of KCs on the membrane potential was analyzed with Eyring rate theory in which Cs+ was assumed to block the K+ transport by binding to a site within the channel. The analysis showed that this site is located at a relative electrical distance delta = 0.32 of the voltage drop across the apical membrane, measured from the cytosolic side. The Hill coefficient obtained from this analysis was n = 3.1. Experiments with K+-depolarized tissues showed that only inward K+ currents recorded with positive transepithelial voltages were depressed by external Cs+. Also under these conditions KCs showed an exponential dependence on the transepithelial potential. The analysis of these data with the rate theory revealed delta = 0.09 and n = 1.7.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1988

Cation-selective channels in amphibian epithelia: electrophysiological properties and activation.

1. Na+ as well as Li+ move across the apical membrane through amiloride-sensitive ionic channels. 2. K+ movements across the apical membrane occur through Ba2+- and Cs+-sensitive channels which do not allow the passage of Na+ or Li+. 3. A third pathway in the apical membrane is permeable for Na+, K+, Cs+, Rb+, NH+4 and Ti+. The currents carried by these monovalent cations are blocked by Ca2+ and divalent cations as well as La3+. 4. In the urinary bladder, the Ca2+-sensitive currents are stimulated by oxytocin, activators of cytosolic cAMP and cAMP analogues. Also the oxytocin activated currents are blocked by divalent cations and La3+. 5. Nanomolar concentrations of mucosal Ag+ activate the third channel and open the pathway for movements of Ca2+, Ba2+ and Mg2+, which are known to permeate through Ca2+ channels in excitable tissues.

Animals↗

PubMed1988

Effect of theophylline on the apical sodium and chloride permeabilities of amphibian skin.

1. The effects of theophylline (1 mmol/l) on the sodium transport (short-circuit current, Isc) and transepithelial conductance (Gtotal) through toad (Bufo viridis) and frog (Rana temporaria and Rana esculenta) skin were investigated. 2. In toad skin incubated with nitrate Ringer solution on the apical side, theophylline induced an increase in Isc similar to that in frog skin bathed with chloride Ringer solution. 3. The increase in Isc could be attributed to recruitment of sodium channels, without affecting the single-channel current. 4. Chloride-bathed toad skin responded to theophylline with a large increase in transepithelial conductance, in addition to the increased Isc. 5. Chloride replacement by nitrate eliminated the effect of theophylline on the conductance increase, but the Isc response was even larger. 6. The results are discussed in relation to the localization of the cellular chloride pathway to the mitochondria-rich cells.

Amphibians↗

PubMed1988

Activation and blockage of a calcium-sensitive cation-selective pathway in the apical membrane of toad urinary bladder.

1. The properties of cation movements through a previously described Ca2+-sensitive oxytocin-stimulated pathway in the apical membrane of the toad urinary bladder were further investigated. 2. In the absence of Ca2+ and other polyvalent cations in the mucosal medium, oxytocin markedly stimulated the flow of current from mucosa to serosa when the major cation in the mucosal solution was any of the following ions: Na+, K+, Rb+, Cs+ or Li+. Analysis of the current noise showed a Lorentzian component associated with the movement of these cations. 3. Ca2+ and other divalent cations in the mucosal solution depressed both the current and the Lorentzian component of the fluctuation spectra. The Michaelis-Menten constants were 2.5, 10 and 58 mumol/l for Ca2+, Sr2+ and Mg2+ respectively. 4. The dihydropyridine Ca2+ channel blockers nitrendipine (10(-5) mol/l) and nicardipine (10(-6) mol/l) inhibited the Ca2+-sensitive current. 5. Alterations of the mucosal pH showed that the current and the plateau of the Lorentzian component increased by elevating the pH from 6 to 8. The Ca2+-sensitive current was further stimulated by increasing pH to 9. However, this manoeuvre resulted in the disappearance of the Lorentzian component in the noise spectrum. 6. Increasing either mucosal [Na+] or [K+] up to 115 mmol/l did not lead to saturation of the current passing through the Ca2+-sensitive channel. In contrast the amiloride-sensitive channel showed saturating behaviour when mucosal [Na+] was increased; half-maximum current was reached when mucosal [Na+] was about 15 mmol/l. 7. When a Na+-free mucosal solution, prepared with either choline or TEA as major cation, was rapidly replaced by a solution with Na+ (115 mmol/l), the current through the Ca2+-sensitive channel increased rapidly and then remained at a nearly constant level. This behaviour is in contrast with the response of the current through the amiloride-sensitive pathway. After suddenly increasing mucosal Na+ concentration ([Na+]m), the current through this channel first increased rapidly and then declined to values of nearly 50% of the peak about 10 min after the increase in [Na+]m.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

PubMed1988

Osmotically induced basolateral K+ conductance in turtle colon: lidocaine-induced K+ channel noise.

The basolateral membrane of amphotericin-treated turtle colon can exhibit two distinct types of K+ conductance, one of which is associated with cell swelling and is blocked by quinidine or lidocaine. Fluctuations in basolateral K+ currents were analyzed under swelling (mucosal KCl) and nonswelling (mucosal K gluconate) conditions. Under nonswelling conditions, it was not possible to detect a spontaneous Lorentzian component in the power density spectrum (PDS) and the addition of lidocaine neither inhibited the macroscopic current nor induced a Lorentzian component in the PDS. Under swelling conditions, however, lidocaine induced a Lorentzian component in the PDS and the corner frequency increased linearly with blocker concentration as expected for reversible blockade of the channel. The gating and conductance properties of osmotically induced channels estimated from a two-state model were similar to those determined recently in single-channel recordings from isolated colonic cells.

Animals↗

PubMed1988

Activation of K+ conductance in basolateral membrane of toad urinary bladder by oxytocin and cAMP.

We incubated toad urinary bladders with Na+-free, isotonic K+ solutions on the apical side and increased the cationic conductance of the apical membrane with nystatin (150 U/ml). Under these conditions, the short-circuit current is mostly carried by K+ flowing from mucosa to serosa. Impedance measurements showed that in nystatin-treated preparations, the electrical behavior of the tissue is dominated by the basolateral membrane properties. Oxytocin (0.1 U/ml) produced an increase of the current and the conductance of the basolateral membrane. Both the resting and the oxytocin-stimulated current were rapidly and reversibly blocked by serosal Ba2+. Addition of the adenosine 3',5'-cyclic monophosphate (cAMP) analogue [8-(4-chloropheylthio)-cAMP] to the basolateral solution mimicked the effects of oxytocin. These results show that oxytocin and cAMP stimulate a potassium conductance in the basolateral membrane and that the stimulation is not related to an increase in sodium entry through the apical membrane. Addition of ouabain (10(-3) M) to the serosal solution did not modify the stimulation by oxytocin, indicating that the activated pathway is not linked to the rate of turnover of the Na+ pump.

Animals↗

PubMed1987

Ca2+ channels in the apical membrane of the toad urinary bladder.

Previously (Van Driessche et al. 1987) we showed that small inward (mucosa towards serosa) oriented short-circuit currents (Isc) were recorded through the toad urinary bladder when the mucosal side was exposed to Ca2+ free solutions containing K+, Na+ (+ amiloride), Cs+ or Rb+ as main cation. This current component is inhibitable by micromolar concentrations of mucosal La3+ and divalent cations (Ca2+, Cd2+) and is considerably elevated by oxytocin (0.1 U/ml). The present study demonstrates that the addition of 50 nmol/l Ag+ to the mucosal medium during oxytocin treatment caused an additional large increase of the La3+-sensitive Isc component. The power density spectrum of the fluctuation in current contained a Lorentzian component which was enhanced by oxytocin treatment. The Lorentzian component disappeared as a consequence of the administration of mucosal Ag+. In experiments with Ca2+, Ba2+ or Mg2+ as principal mucosal cation, the La3+-sensitive Isc component was negligible under control conditions and during oxytocin treatment. Mucosal Ag+ (40 nmol/l) elicited a large inward oriented current which was blockable by the calcium channel blockers, La3+ and Cd2+. Also the organic calcium entry blockers, nicardipine and verapamil (10 mumol/l) depressed the inward current considerably. Noise analysis of the currents carried by divalent cations showed a La3+-sensitive noise component. Oxytocin-Ag+ activated currents could not be recorded in the absence of the divalent cations or small inorganic cations, e.g. with solutions which contained N-methyl D-glucamine (NMDG) as main mucosal cation.

Animals↗

PubMed1987

Oxytocin and cAMP stimulate monovalent cation movements through a Ca2+-sensitive, amiloride-insensitive channel in the apical membrane of toad urinary bladder.

The effects of oxytocin and cAMP on ion transport were investigated in toad urinary bladders incubated with Ca2+-free solutions on the apical side. Under these conditions both oxytocin and cAMP markedly stimulated the movements of Na+, K+, Rb+, Cs+, Li+, and NH4+ through a pathway that is insensitive to amiloride. The amiloride-insensitive currents were inhibited by the addition of Ca2+, Sr2+, or Mg2+ to the apical solution. The movement of the monovalent cations was associated with a spontaneous Lorentzian component in the power spectrum of the fluctuation in short-circuit current. The plateau of the Lorentzian component was enhanced by oxytocin and cAMP and was depressed by divalent cations. Methohexital inhibited the stimulation of monovalent cation movements caused by oxytocin. These findings suggest that oxytocin and cAMP activate at least two kinds of ionic channels in the apical membrane of toad urinary bladder: the well-known amiloride-sensitive channel and an amiloride-insensitive channel that allows the movement of several monovalent cations and is blocked by Ca2+ and other divalent cations.

Amiloride↗

PubMed1986

Impairment of Na+ transport across frog skin by Tl+: effects on turnover, area density and saturation kinetics of apical Na+ channels.

Na+ transport across abdominal skin of the frogs, Rana temporaria and Rana esculenta was followed by measuring Na+-dependent short-circuit, current (INa) kinetics and INa fluctuations induced by triamterene, a diuretic. Exposure of the skin to serosal Tl+ led to a pronounced and irreversible drop in INa and INa-blocker noise. At low serosal Tl+ concentrations, we observed mainly a decrease in the apparent Michaelis constant for INa saturation while, at larger [Tl+], the maximal INa dropped irreversibly. Tl+ acts even when serosal Tl+ "transporters" like the Na+-K+ pump, or the K+ channel are nonfunctional. The rate constants for the triamterene/Na+ channel reaction were unchanged after Tl+ whereas the relaxation noise from channel blockage decreased in amplitude. Noise analysis in terms of a two-state blocking model suggested that Tl+ poisoning results in a small decrease in single-channel current through apical Na+ pathways, as well as in a drastic and irreversible drop in channel density. The impairment of Na+ transport by Tl+ can be attributed to the above cited concerted events at the level of the apical membrane.

Animals↗