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W Van Driessche

Publications and source records attributed to W Van Driessche.

At least 109 records · Page 6Linked to original sources

Noise analysis reveals K+ channel conductance fluctuations in the apical membrane of rabbit colon.

In this paper we describe current fluctuations in the mammalian epithelium, rabbit descending colon. Pieces of isolated colon epithelium bathed in Na+ or K+ Ringer's solutions were studied under short-circuit conditions with the current noise spectra recorded over the range of 1-200 Hz. When the epithelium was bathed on both sides with Na+ Ringer's solution (the mucosal solution contained 50 microM amiloride), no Lorentzian components were found in the power spectrum. After imposition of a potassium gradient across the epithelium by replacement of the mucosal solution by K+ Ringer's (containing 50 microM amiloride), a Lorentzian component appeared with an average corner frequency, fc = 15.6 +/- 0.91 Hz and a mean plateau value So = (7.04 +/- 2.94) x 10(-20) A2 sec/cm2. The Lorentzian component was enhanced by voltage clamping the colon in a direction favorable for K+ entry across the apical membrane. Elimination of the K+ gradient by bathing the colon on both sides with K+ Ringer's solutions abolished the noise signal. The Lorentzian component was also depressed by mucosal addition of Cs+ or tetraethylammonium (TEA) and by serosal addition of Ba2+. The one-sided action of these K+ channel blockers suggests a cellular location for the fluctuating channels. Addition of nystatin to the mucosal solution abolished the Lorentzian component. Serosal nystatin did not affect the Lorentzian noise. This finding indicates an apical membrane location for the fluctuating channels. The data were similar in some respects to K+ channel fluctuations recorded from the apical membranes of amphibian epithelia such as the frog skin and toad gallbladder. The results are relevant to recent reports concerning transcellular potassium secretion in the colon and indicate that the colon possesses spontaneously fluctuating potassium channels in its apical membranes in parallel to the Na+ transport pathway.

Animals↗

The effect of lanthanum on alamethicin channels in black lipid bilayers.

The properties of alamethicin channels in dioleyl phosphatidylcholine bilayers were studied in 1 M LaCl3 and were compared with those in 1 M NaCl. Single-channel recordings demonstrated that the mean single-channel life-time is about 0.25 s in NaCl but only about 17 ms in LaCl3. Whereas in NaCl the conductance levels 2 and 3 are mostly populated, in LaCl3 the levels 0 and 1 are preferentially adopted. The single-level conductance are slightly smaller in LaCl3 if the higher bulk solution conductivity of LaCl3 is taken into account. Multipore experiments confirmed earlier results (Boheim, G., Irmscher, G. and Jung, G. (1978) Biochim. Biophys. Acta 507, 485--506) that the bilayer conductance is less strongly dependent on voltage in LaCl3 than in NaCl solution. Current-fluctuation analysis showed that this effect can be explained by a less strong dependence on voltage of the pore-formation rate as well as of the mean channel life-time in LaCl3. The data can be interpreted as an increased lateral diffusion mobility of the alamethicin monomers in the bilayer. This can be the result of the binding of La3+ to the polar headgroups which can induce cluster formation of the phospholipids.

Alamethicin↗

Capacitive and inductive low frequency impedances of Necturus gallbladder epithelium.

The electrical impedance of Necturus gallbladder epithelium was analysed in the frequency range 0.24 Hz to 6,323 Hz. Under control conditions (NaCl-Ringer's on both sides), the impedance function yields a semicircle with depressed center. When serosal Na+ was replaced by K+, an inductive low frequency (LF) component appeared in the impedance locus. With KCl-Ringer on the mucosal side a second circular arc was observed at frequencies below 1 Hz. The resistive parts of the capacitive and inductive LF components increased after application of TAP+ to the mucosal side. Both LF features were abolished after application of 5 mM TEA+ to the mucosal medium as well as after acidification of the mucosal side. The LF components were depressed by addition of 5 mM Ba2+ to the mucosal solution. As TEA+ blocks apical K+ channels (Van Driessche and Gögelein 1978), it is concluded that the capacitive as well as the inductive LF components are related to transcellular K+ flow. With KCl-Ringer on the mucosal side, mucosa negative potentials increased the equivalent resistance and decreased the equivalent capacitance of the LF impedance. With serosal KCl-Ringer, negative potentials evoked a capacitive component which overlapped with the inductive component observed at open circuit conditions. Positive potentials, however, abolished the capacitive as well as the inductive LF component, elicited by mucosal or serosal KCl-Ringer, respectively. These results demonstrate that serosa to mucosa directed K+ flow causes an inductive LF feature and that mucosa negative potentials elicit a capacitive LF component.

Animals↗

Apical K+ channels in frog skin (Rana temporaria): cation adsorption and voltage influence gating kinetics.

Open-close kinetics of fluctuating K+ channels in the apical frog skin membrane were studied with noise analysis of the K+ current (IK). The mucosa to serosa directed IK was obtained with serosal NaCl- and mucosal KCl-Ringer under voltage clamp conditions. Mucosal protons (pH greater than 4), several polyvalent metal ions, and choline shifted the plateaus (SO) of the Lorentzian component in the IK noise spectrum to higher, but the corner frequency (fc) to lower values. SO was lowered at pH less than 4, due to a K+-channel block by H+. Ca2+, Sr2+, H+ (pH greater than 4) and choline did not affect IK. A slight reduction of IK was seen with Mg2+, Mn2+, Co2+, Ni2+, Zn2+, Cu2+ and La3+. At pH greater than 4, the H+-induced shifts in SO an fc were almost abolished in solutions of high mucosal Ca2+ concentrations. Clamping the transepithelial potential difference to more positive values (with respect to the serosa) shifted the Lorentzian parameters SO and fc in the same way as the cations did. As with protons, mucosal Ca2+ interferred with the effect of voltage. The interference of cationic (probably fixed charge screening) and voltage effects suggests a common, more general mechanism of action, namely alterations in K+-channel fluctuation kinetics by changes in local electrical fields. On this basis, the rates for the open-close reaction of K+ channels and their mean lifetime were calculated. We found that e.g. increasing [Ca2+]O from 1-10 mM caused no change of the mean open time, but increased the mean time "closed" of the K+ channel by a factor of about 1.5. Other mucosal cations, as well as depolarizing clamp potentials are thought to have the same effect.

Animals↗

Noise analysis of the K+ current through the apical membrane of Necturus gallbladder.

Current noise power spectra of the voltage-channel (V = 0) Necturus gallbladder, exposed to NaCl-Ringer's on both sides contained a relaxation noise component, which overlapped with a 1/f alpha noise component, with alpha being about 2. Substitution of all Na+ by K+ on either the serosal or mucosal side increased the relaxation as well as the 1/f alpha noise component considerably. In Necturus gallbladder both noise components are reduced by addition of 10 mM, 2,4,6-triaminopyrimidine (TAP) or 5 mM BA2+ to the mucosal side, as well as by acidification of the mucosal solution to pH 5 and lower. Five mM of tetraethylammonium (TEA+) added to the mucosal solution, abolished K+ relaxation noise and decreased the 1/f alpha noise component. Applying a Cs+ concentration gradient across the epithelium did not yield relaxation noise. However, if Rb+ was substituted for all Na+ on one side, a Lorentzian noise component appeared in the spectrum. Its plateau was smaller than with KCl-Ringer's on the respective side. These data confirm the existence of fluctuating K+ channels in the apical membrane of the Necturus gallbladder. Furthermore it can be concluded that these channels have the permeability sequence K+ greater than Rb+ greater than Cs+. The inhibition of the fluctuations by mucosal acidification indicates the existence of acidic sites in the channel. The single-channel conductance was estimated to be between 6.5 and 40 pS.

Animals↗

Noise analysis of the K+ current through the apical membrane of Necturus gallbladder.

Current noise power spectra of the voltage-clamped (V = 0) Necturus gallbladder, exposed to NaCl-Ringer's on both sides contained a relaxation noise component, which overlapped with a 1/f alpha noise component, with alpha being about 2. Substitution of all Na+ by K+ on either the serosal or mucosal side increased the relaxation as well as the 1/f alpha noise component considerably. In Necturus gallbladder both noise components are reduced by addition of 10mM 2,4,6-triaminopyrimidine (TAP) or 5 mM of tetraethylammonium (TEA+) added to ification of the mucosal solution to pH 5 and lower. Five mM of tetraethylammonium (TEA+) added to the mucosal solution, abolished K+ relaxation noise and decreased the 1/f alpha noise component. Applying a Cs+ concentration gradient across the epithelium did not yield relaxation noise. However, if Rb+ was substituted for all Na+ on one side, a Lorentzian noise component appeared in the spectrum. Its plateau was smaller than with KCl-Ringer's on the respective side. These data confirm the existence of fluctuating K+ channels in the apical membrane of the Necturus gallbladder. Furthermore it can be concluded that these channels have a permeability sequence K+ greater than Rb+ greater than Cs+. The inhibition of the fluctuations by mucosal acidification indicates the existence of acidic sites in the channel. The single-channel conductance was estimated to be between 6.5 and 40 pS.

Animals↗

Ba2+-induced conductance fluctuations of spontaneously fluctuating K+ channels in the apical membrane of frog skin (Rana temporaria).

We studied the influence of mucosal Ba2+ ions on the recently described (Zeiske & Van Driessche, 1979a, J. Membrane Biol. 47:77) transepithelial, mucosa towards serosa directed K+ transport in the skin of Rana temporaria. The transport parameters G (conductance), PD (potential difference), Isc (short-circuit current, "K+ current"), as well as the noise of Isc were recorded. Addition of millimolar concentrations of Ba/+ to the mucosal K+-containing solution resulted in a sudden but quickly reversible drop in Isc. G and Isc decreased continuously with increasing Ba2+ concentration, (Ba2+)o. The apparent Michaelis constant of the inhibition by Ba2+ lies within the range 40-80 microM. The apical membrane seems to remain permselective for K+ up to 500 microM (Ba2+)o. Higher (Ba2+)o, however, appears to induce a shunt (PD falls, G increases). This finding made an accurate determination of the nature of the inhibition difficult but our results tend to suggest a K+-channel block by K+-Ba2+ competition. In the presence of Ba2+, the power spectrum of the K+ current shows a second Lorentzian component in the low-frequency range, in addition to the high-frequency Lorentzian caused by spontaneous K+-channel fluctuations (Van Driessche & Zeiske, 1980). Both Lorentzian components are only present with mucosal K+ and can be depressed by addition of Cs+ ions, thus indicating that Ba2+ ions induce K+-channel fluctuations. The dependence of the parameters of the induced Lorentzian on (Ba2+)o shows arise in the plateau values to a maximum around 60 microM (Ba2+)o, followed by a sharp and progressive decrease to very low values. The corner frequency which reflects the rate of the Ba2+-induced fluctuations, however, increases quasi-linearly up to 1 mM (Ba2+)o with a tendency to saturate at higher (Ba2+)o. Based on a three-state model for the K+ channel (having one open state, one closed by the spontaneous fluctuation and one blocked by Ba2+) computer calculations compared favorably with our results. The effect of Ba2+ could be explained by assuming reversible binding at the outer side of the apical K+ channel, thereby blocking the open channel in ;competition with K+. The association-dissociation of Ba2+ at its receptor site is thought to cause a chopping of the K+ current, resulting in modulated current fluctuations.

Action Potentials↗

Spontaneous fluctuations of potassium channels in the apical membrane of frog skin.

1. The previously demonstrated K+-dependent short-circuit current through the skin of frog species Rana temporaria (Zeiske & Van Driessche, 1979), bathed with mucosal K+- and serosal Na+-Ringer solution, was investigated with current-fluctuation analysis. 2. The current-noise spectra were recorded in the frequency range from 1 to 800 Hz and showed a Lorentzian component with a mean plateau value S0 = (1.50 +/- 0.05).10(-20) A2.s.cm-2 and a corner frequency of fc=(81.0 +/- 3.4)Hz(n=14). 3. S0 increased with mucosal K+ concentration, [K]o, while fc remained almost unchanged. A decrease in S0 was observed when serosal Na+ was replaced by K+. 4. Mucosal Cs+ (10 mM) depressed, reversibly, the K+-dependent current noise to the level of the background noise. Moreover, a linear decrease in fc with increasing Cs+ concentration was observed. 5. Among the other tested alkali cations, Rb+ was the only blocker though less potent than Cs+. Tetraethylammonium, 4-aminopyridine, 2.4.6-triaminopyrimidine and amiloride had no effect. 6. Alterations in the transcellular transport of Na+ contained in a mucosal solution with high [K]o resulted in significant changes in K+ current noise. 7. The current-fluctuation intensities decreased with increasing contact time to high [K]o; these changes were concomitant with the previously reported time dependence of the short-circuit current (Zeiske & Van Driessche, 1979). 8. The K+-dependent fluctuations are thought to originate from K+-selective pathways in the apical cell membranes. The description of the K+-current noise by a single Lorentzian suggests that the "K+ channels" switch randomly between an open and closed state. 9. Assuming a two state model for the channel-kinetics, the single channel current i and the channel density M were calculated as i=(0.37 +/- 0.05)pA and M=(0.53 +/- 0.08) mu-2 (n=13).

Animals↗

Saturable K+ pathway across the outer border of frog skin (rana temporaria): kinetics and inhibition by Cs+ and other cations.

The reaction of abdominal skins of the frog species Rana temporaria on mucosal K+-containing solutions was studied in an Ussing-type chamber by recording transepithelial potential difference (PD), short-circuit current (SCC) and conductance (G). With Na-Ringer's as serosal medium, a linear correlation between PD and the logarithm of the mucosal K+-concentration ([K]o) was obtained. The K+-dependent SCC saturated with increasing [K]o, and could quickly and reversibly be depressed by addition of Rb+, Cs+, and H+. Li+, Na+, and NH4+ did not influence K+ current. A large scatter was obtained for kinetic parameters like the slope of the PD-log[K]o-line (18--36.5 mV/decade), the apparent Michaelis constant (13--200 mM), and the maximal current of the saturable SCC (6--50 microa . cm-2), as well as for the degree of inhibition by Cs+ ions. This seemed to be caused by a time-dependent change during long time exposure to high [K]o (more than 30 sec), thereby inducing a selectivity loss of K+-transporting structures, together with an increase in SCC and G and a decrease in PD. Short time exposure to K+-containing solutions showed a competitive inhibition of K+ current by Cs+ ions, and a Michaelis constant of 6.6 mM for the inhibitory action of Cs+. Proton titration resulted in a decrease of K+ current at pH less than 3. An acidic membrane component (apparent dissociation constant 2.5 x 10(-3) M) is virtually controlling K+ transfer. Reducing the transepithelial K+-concentration gradient by raising the serosal potassium concentration was accompanied by the disappearance of SCC and PD.

Animals↗