Effect of forskolin on conductive anion pathways of toad skin

The effect of the diterpene, forskolin, on pathways for conductive Cl- transport was analyzed using isolated skins of Bufo viridis. Forskolin did not stimulate the voltage-activated Cl- movement from mucosa to serosa; the Lorentzian component in the power density spectrum, which was present at serosa positive clamp potentials under control conditions, decreased significantly. The observation that stimulation of cytosolic adenosine 3'-5'-cyclic monophosphate (cAMP) by forskolin has no effect on the voltage-activated Cl- transport argues against control of this pathway by cAMP. Our data further demonstrate that the forskolin-activated Cl- conductive pathway is also permeable for NO3-. This pathway was studied in absence of mucosal Cl-, which eliminates Cl- movement through the voltage-activated pathway. With SO4(2-) and Cl- on the mucosal and serosal sides, respectively, this forskolin-induced pathway displayed a linear current-voltage relationship. The associated Lorentzians increased at serosa negative clamp potentials. Transepithelial current and plateau value of the Lorentzian were related by a quadratic function, which suggests voltage-independence of number and open-close probability of these conductance sites. Morphological sites for voltage-activated and forskolin-induced conductive Cl- transport remain to be identified.

Effect of mucosal halides on Ca(2+)-blockable currents through the skin of Rana ridibunda

The present study deals with the interaction of mucosal anions with apical Ca(2+)-blockable cation channels of the skin of Rana ridibunda. The intracellular potential was depolarized by exposing the basolateral membranes to K2SO4 Ringer solution. The apical bathing medium consisted of nominal Ca(2+)-free K+ or Na+ solutions with SO4(2-), Cl-, Br-, or I- as the major anion. The effects of mucosal anion substitutions were studied by analyzing 1) the fluctuation in K+ current across the apical membrane driven by imposed transepithelial clamping potentials and 2) alterations of the transepithelial current (It) and conductance (Gt) as well as the Lorentzian parameters in response to anion substitution in the mucosal bathing solution. It and current noise spectra were recorded at different transepithelial potentials (Vt). A Lorentzian component was present in the power density spectrum when Vt was clamped at mucosa-positive voltages. Such noise components were never observed with mucosa-negative potentials. These findings suggest a rectifying behavior of the transepithelial cation currents. The Lorentzian noise component and the inward-oriented cation currents were depressed by the addition of micromolar concentrations of Ca2+ to the apical solutions as well as by replacing mucosal K+ or Na+ by N-methyl-D-glucamine. The Ca(2+)-blockable current and Lorentzian noise plateau (So) were gradually increased by raising Vt. Both parameters, as well as the corner frequency (fc), depended strongly on the major anion species in the apical solution; replacing mucosal SO4(2-) by one of the halides tested reduced fc and elevated So, It, and Gt considerably.

K+ secretion across frog skin. Induction by removal of basolateral Cl-.

We examined the development of K+ secretion after removing Cl- from the basolateral surface of isolated skins of Rana temporaria using noise analysis. K+ secretion was defined by the appearance of a Lorentzian component in the power density spectrum (PDS) when Ba2+ was present in the apical bath (0.5 mM). No Lorentzians were observed when tissues were bathed in control, NaCl Ringer solution. Replacement of basolateral Cl- by gluconate, nitrate, or SO4- (0-Clb) yielded Lorentzians with corner frequencies near 25 Hz, and plateau values (So) that were used to estimate the magnitude of K+ secretion through channels in the apical cell membranes of the principal cells. The response was reversible and reproducible. In contrast, removing apical Cl- did not alter the PDS. Reduction of basolateral Cl- to 11.5 mM induced Lorentzians, but with lower values of So. Inhibition of Na+ transport with amiloride or by omitting apical Na+ depressed K+ secretion but did not prevent its appearance in response to 0-Clb. Using microelectrodes, we observed depolarization of the intracellular voltage concomitant with increased resistance of the basolateral membrane after 0-Clb. Basolateral application of Ba2+ to depolarize cells also induced K+ secretion. Because apical conductance and channel density are unchanged after 0-Clb, we conclude that K+ secretion is "induced" simply by an increase of the electrical driving force for K+ exit across this membrane. Repolarization of the apical membrane after 0-Clb eliminated K+ secretion, while further depolarization increased the magnitude of the secretory current. The cell depolarization after 0-Clb is most likely caused directly by a decrease of the basolateral membrane K+ conductance. Ba2(+)-induced Lorentzians also were elicited by basolateral hypertonic solutions but with lower values of So, indicating that cell shrinkage per se could not entirely account for the response to 0-Clb and that the effects of 0-Clb may be partly related to a fall of intracellular Cl-.

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.

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.

Cl- - and K+-related fluctuations of ionic current through oxyntic cells in frog gastric mucosa

Transport and conductance pathways for Cl- and K+ were studied in frog gastric mucosa using noise analysis techniques. The current-noise power spectra exhibited both Cl- - and K+-dependent characteristics. In Cl- -containing solutions, reductions in Cl- transport were associated with reductions of the overall noise power. Such changes appeared to reflect the movement of Cl- through the apical (mucosal) membranes of oxyntic cells. In Cl- -free solutions a K+-dependent Lorentzian component was detected in the power spectrum when applying a mucosally or serosally directed transepithelial K+ concentration gradient. This component was enhanced by 1) stimulating the oxyntic cells with histamine and 2) appropriate voltage clamping. It was reduced by mucosal Ba2+ in resting tissues and enhanced by mucosal Ba2+ in stimulated tissues. The K+ noise measured in gastric mucosae in Cl- -free solutions appeared also to be generated at the apical membranes of oxyntic cells. This is in analogy to previous findings in other gastrointestinal epithelia with fluctuating apical K+ channels. In the gastric mucosa these channels may play a key role in the mechanism of electrogenic H+ secretion.

The Resonant Faraday Effect

The resonant Faraday effect occurs for radiation at frequencies close to the absorption lines of a magneto-active material. In the paper we present a theory for isolated absorption lines with Voigt profiles. The background source is also assumed to have a Voigt profile. The theory shows how the resonant Faraday effect can be used to determine relative transition probabilities, as well as the magnitude of the Lorentzian component of the line profiles.Experimental results for an argon plasma are presented and show that the relative transition probabilities of atomic transitions with a common lower level can be measured to an accuracy of ± 1%. For strong absorption lines, the Voigt "a" parameter can also be determined to an accuracy of 10%, even when the Doppler width is 14 times larger than the Lorentzian width.

Self broadening and f -values in the spectrum of neon

Profiles of lines in the NeI spectrum arising from transitions between the configurations 2 p 5 3 s and 2 p 5 3 p were measured by means of a direct-recording Fabry–Perot interferometer. The d. c. discharge was cooled with liquid nitrogen or ice and currents were typically 0·5 mA. Full analysis of the profiles determines the pressure broadening as a Lorentzian component. For lines with the lower level 2 s 2 (Paschen notation) the broadening can be ascribed to the resonance interaction alone and the f -value of 0·16 8 is obtained for the resonance transition 736 Å. From coupling parameters the f -value of the transition 744 Å is estimated as 0·01 2 which is consistent with the broadening observed for the lines with the lower level 2 s 4 . The nature of the higher order interactions is discussed with reference to the observed temperature dependence and pressure shifts. Extrapolation of the Lorentzian width to zero density and the subtraction of a carefully determined instrumental contribution gives a value in excess of the radiation width for the level 2 s 2 , a result similar to that reported for the level 2 1 P 1 in helium. The explanation of this anomaly as an enhancement effect is discussed.