Biomedical subjects

W Nagel

Publications and source records attributed to W Nagel.

At least 73 records · Page 4Linked to original sources

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

Chloride conductance of frog skin: localization to the tight junctions?

The pathway(s) of passive conductive Cl transport across isolated frog skin are analyzed by electrophysiological techniques including microelectrode impalement of principal cells. It is found that the apical membrane of granular cells is virtually impermeable for Cl. Substitution of mucosal Cl by anions except NO3 and SCN decreases Cl-related tissue conductance (gCl) with first order kinetics. NO3 and SCN block gCl with half-maximal concentration of 18 and 5 mM, respectively. Omission of serosal Cl has little effect on gCl unless the inhibiting anions NO3 or SCN are used. The putative Cl channel blocker diphenylaminocarbonic acid (DPC) and some analogs inhibit gCl, half-inhibitory concentration of the most potent dichloro-DPC is 10(-6) M. The inhibitors act only from the mucosal side. Slow dissipation of gCl after abolition of Na entry across the apical membranes which can be prevented by preceding blockage of the Na-K-ATPase with ouabain suggests that the intracellular Na activity might influence the permeability of the Cl pathway. It is supposed that this control involves microfilaments between the cytoskeleton and the tight junctions with Cl-specific permeation sites in outer regions of the junctional complex.

Amiloride↗

PubMed1989

Glomerular tubular balance: mediation by luminal hypotonicity.

Late proximal rat tubular segments were microperfused with slightly hypo- or hypertonic artificial late proximal tubular fluid (ATF) at low (11-13 nl/min) or high (30-38 nl/min) perfusion rates. Volume reabsorption, net chloride and solute reabsorption were measured as a function of length. In addition, the transepithelial resistance and voltage (Vte) were measured as a function of the applied osmotic gradient. Hypertonic solutions equilibrated to isotonicity by solute outflow rather than water influx. With hypertonic ATF the lumen positive Vte was decreased compared with free flow or with hypotonic ATF. The resistance was not significantly different between the different groups. In contrast to hypotonic ATF, hypertonic or isotonic ATF was not significantly reabsorbed. In addition, hypotonic ATF maintained its hypotonicity along the perfused segments. Its reabsorption was flow-dependent. Hypotonicity appeared to enhance solute reabsorption.

Absorption↗

PubMed1988

Effects of divalent cations on chloride movement across amphibian skin.

Effects of the divalent heavy metal ions Cd2+, Co2+, Cu2+, Mn2+, Ni2+, and Zn2+ on pathways for sodium and chloride were assessed on isolated amphibian skin (Rana temporaria and esculenta, Bufo marinus and viridis). It was observed that these agents, in addition to the previously reported stimulation of sodium transport, inhibit chloride-related tissue conductance (gCl) in frog skin with spontaneously high gCl when added to the external incubation medium. Serosal application was ineffective. Half-maximal inhibition of gt occurred at approximately 0.2 mmol/l Ni2+ and Zn2+, 0.5 mmol/l Co2+ and Cd2+, and more than 3 mmol/l Mn2+. The onset of inhibition was rapid, steady state values being reached within 3 min; reversibility was complete with approximately similar time course. Cu2+, which could not be tested at concentrations above 0.1 mmol/l, had only minimal and poorly reversible effect on gCl. Skin of Bufo was virtually insensitive to these metal ions. Microelectrode determinations demonstrate that the decrease of conductance was restricted to a pathway distinct from the principal cells which show, on the contrary, increase of apical membrane conductance originating from stimulation of sodium permeability. The metal ions might be valuable for characterization of the pathway and the mechanism of transepithelial conductive chloride transport.

Animals↗

PubMed1988

Uptake of Br in mitochondria-rich and principal cells of toad skin epithelium.

To elucidate the route of transepithelial Cl transport across amphibian skins, electrolyte concentrations and uptake of Br in different epithelial cell types of toad skin were determined using electron microprobe analysis. Under short-circuited conditions, Cl concentrations were about 10 mmol/kg ww lower in MR-cells (23.9 +/- 9.6 mmol/kg ww) than in principal cells and showed a large scatter. After unilateral substitution of Br for Cl in the bathing solutions, principal cells exchanged Br for Cl only from the serosal side, whereas variable amounts of Br were gained in MR-cells from either side. The ratio of Br to Cl concentrations in MR-cells averaged 0.35 and 0.81 after incubation with NaBr-Ringer's on the apical or serosal side, respectively. After activation of transepithelial anion conductance by serosa-positive voltage-clamping to 100 mV, uptake of Br from the apical side was increased in MR-cells compared with short-circuited conditions. On the average, the ratio of cellular Br to Cl concentrations was 1.38, but the variation among individual MR-cells from the same tissue was considerable. In MR-cells with large uptake of Br and voltage-activated conditions, the sum of Br and Cl concentrations was higher than the Cl concentration under control conditions. The increase of anion content was associated by increase of the Na and corresponding decrease of the K concentrations. The MR-cells were swollen as indicated by the decrease in the cellular dry weight content from 22.2 +/- 2.5 to 17.1 +/- 4.2 g/100 g.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1988

Procaine effects on sodium and chloride transport in frog skin.

Procaine, a tertiary amine, has previously been shown to stimulate reversibly transepithelial Na transport across frog skin after application from the epithelial side. In the present study with intracellular recording from principal, i.e. amiloride-sensitive cells, we demonstrate that the stimulation results from increase in apical membrane Na permeability. A second effect of procaine (10-25 mmol/l) in the outside perfusion solution is a reversible increase of transepithelial conductance which drastically exceeds the predicted response of the transcellular Na pathway. It requires presence of chloride on the epithelial side and depends on the non-ionized molecule of procaine. Abolition of apical membrane Na uptake by amiloride or Na-free mucosal incubation decreases the magnitude but does not prevent the stimulatory effect of procaine. The origin of this gain in conductance from stimulation of a Cl-specific pathway is demonstrated by a highly significant correlation between the increases in electrically determined tissue conductance and partial Cl conductance, obtained from measurements of influx and efflux of Cl-36. Measurements with microelectrodes indicate that the stimulated Cl-specific pathway is distinct from the principal cells. Since procaine activates a conductive pathway with similar response pattern as spontaneously existing Cl conductance, it might be a valuable tool for investigating mode and way of Cl movement across epithelial tissues.

Animals↗

PubMed1988

Voltage dependence of cellular current and conductances in frog skin.

Knowledge of the voltage dependencies of apical and basolateral conductances is important in determining the factors that regulate transcellular transport. To gain this knowledge it is necessary to distinguish between cellular and paracellular currents and conductances. This is generally done by sequentially measuring transepithelial current/voltage (It/Vt) and conductance/voltage (gt/Vt) relationships before and after the abolition of cellular sodium transport with amiloride. Often, however, there are variable time-dependent and voltage-dependent responses to voltage perturbation both in the absence and presence of amiloride, pointing to effects on the paracellular pathway. We have here investigated these phenomena systematically and found that the difficulties were significantly lessened by the use of an intermittent technique, measuring It and gt before and after brief (less than 10 sec) exposure to amiloride at each setting of Vt. I/V relationships were characterized by these means in frog skins (Rana pipiens, Northern variety, and Rana temporaria). Cellular current, Ic, decreased with hyperpolarization (larger serosa positive clamps) of Vt. Derived Ic/Vt relationships between Vt = 0 and 175 mV (serosa positive) were slightly concave upwards. Because values of cell conductance, gc, remained finite, it was possible to demonstrate reversal of Ic. Values of the reversal potential Vr averaged 156 +/- 14 (SD, n = 18) mV. Simultaneous microelectrode measurements permitted also the calculation of apical and basolateral conductances, ga and gb. The apical conductance decreased monotonically with increasing positivity of Vt (and Va). In contrast, in the range in which the basolateral conductance could be evaluated adequately (Vt less than 125 mV), gb increased with more positive values of Vt (and Vb). That is, there was an inverse relation between gb and cellular current at the quasi-steady state, 10-30 sec after the transepithelial voltage step.

Amiloride↗

PubMed1988

[Immunocytochemical detection of the estrogen receptor and lectin binding sites in tumor cell colonies growing in vitro].

Since it became clear that tumor cell colonies growing in soft agar are closely related to embryonic tumor cells, it has been possible to examine, with the aid of immunohistochemical methods, the behavior of estrogen receptors (ER) and lectin binding sites on those tumor components which determine malignant growth. With this aim in mind, 14 breast cancers, four ovarian cancers, and one corpus cancer with ER concentrations known from DCC studies were cultured in a colony assay. Using immunohistochemical techniques it was possible, after preparing permanent cultures, to demonstrate the estrogen receptors and lectin binding sites on the tumor cell colonies cultured in vitro.

Breast↗

PubMed1988

Temperature dependence of transcellular and intracellular parameters of frog skin.

Studies on the mechanism(s) for temperature dependence of sodium transport in tight epithelia reported in literature, have primarily been derived from transcellular studies. No attempts were made to discriminate between trans and paracellular conduction pathways nor is it possible from such measurements to examine the contribution of the apical and/or basolateral cellular membranes to the overall temperature dependency of the transport process. In this study we report on the temperature dependence of intracellular as well as transcellular parameters. Using a specific protocol, which provides the temperature coefficients for the amiloride displaceable sodium current, cellular conductance and the apical and basolateral membrane, conductances. Temperature coefficients derived from the slopes of Arrhenius plots of these parameters were found to be very similar (9-10 KCal/mole). The intracellular potential before and after amiloride showed weak temperature dependence (less than 1 mV/degrees C), however. On the basis of our results here, and in view of certain theoretical considerations, we conclude that the inhibition of Na+ transport across frog skin by decreasing the temperature is primarily due to a decrease in the permeability of the apical and basolateral membranes. This decrease in permeability is brought about by changes in the viscosity of water and the radius of the moving particle; in this case sodium plus hydration shell. The effect of temperature on the intracellular potential could be partially accounted for by changes in the equilibrium-potential for potassium and in part to changes in the rheogenic transport component for sodium across the basolateral membrane.

Amiloride↗

PubMed1988