PubMed Health⌕ Search

Biomedical subjects

S Krasne

Publications and source records attributed to S Krasne.

14 recordsLinked to original sources
PubMed1984

Two calcium currents in Neanthes arenaceodentatus egg cell membranes.

Two distinct types of Ca currents, Ca(I) and Ca(II), were found in the eggs of the marine polychaete Neanthes arenaceodentatus and studied under voltage-clamp conditions. Ca(I) and Ca(II) channels differ in their selectivity sequences, show different sensitivities to blocking by Cd, and have different activation thresholds. These facts indicate that the channels responsible for Ca(I) and Ca(II) currents are unique and different. Both Ca(I) and Ca(II) currents decrease with time under a maintained depolarization. This relaxation exhibits different kinetics, with those for Ca(II) being an order of magnitude slower than for Ca(I). The Ca(I) current relaxation has been shown previously to be due to a voltage-dependent inactivation. The magnitude of relaxation of the Ca(II) current elicited by a test-voltage step immediately following a conditioning-voltage step paralleled the magnitude of the peak of the Ca(II) current flowing during the conditioning pulse. The kinetics of the current relaxation depended upon bath Ca concentration, the kinetics slowing down as the bath Ca was increased. These observations are consistent with an external depletion being the cause of the current relaxation for the Ca(II) channel.

Animals↗
PubMed1984

Membrane patches and whole-cell membranes: a comparison of electrical properties in rat clonal pituitary (GH3) cells.

A comparison has been made between the electrical properties of excised 'outside-out' patches and whole-cell membranes of GH3 cells using the patch-pipette technique. Despite a complicated surface morphology, which includes numerous microvilli, ruffles and blebs, high-resistance seals (typically greater than 10(11) omega) were consistently formed between patch pipettes and GH3 cell membranes. When the internal solution contained 120 mM-CsF, outward currents through K channels were blocked and large Na channel currents were consistently observed in the whole-cell recording mode. Using the same solutions, single Na channel currents were readily observed in outside-out patches. Averaging patch currents yielded macroscopic currents showing the same voltage-dependent kinetics as those observed for the whole-cell membrane. The current vs. voltage and inactivation time constant vs. voltage relationships for the Na channel shifted towards more negative potentials (25 mV or more) within approximately 30 min after going into the whole-cell recording mode. These same relationships could be measured for outside-out patches and their positions along the voltage axis coincided with the asymptotic values measured in the whole-cell mode. When the internal solution contained 120 mM-N-methylglucamine fluoride and the external solution contained 120 mM-Tris chloride, no ionic channel currents could be observed either for whole-cell or outside-out patch membranes. Under these conditions, displacement currents induced by tetraphenyl borate (TPB) were recorded in both types of membranes. The total charge moved showed a sigmoidal dependence upon the applied voltage for both whole-cell and outside-out patch membranes. The charge vs. voltage relationship showed a shift along the voltage axis similar to that observed for Na channels except that the magnitude of the shift was larger. A shift in this relationship was also observed for excised patches but the observable magnitude of the shift was smaller than that in the whole-cell recording mode. The asymptotic values of the charge vs. voltage relationship were similar for whole-cell and outside-out patches, as were the asymptotic values for the translocation time constants. It is concluded that there are no fundamental differences in the properties of ionic channel and displacement currents between whole-cell membranes and excised membrane patches.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed1983

Interactions of voltage-sensing dyes with membranes. III. Electrical properties induced by merocyanine 540.

The effects of merocyanine 540 on the electrical properties of lipid bilayer membranes have been investigated. The alterations this dye was found to produce in the intrinsic conductances of these membranes were minimal, but it profoundly altered the conductances produced by extrinsic permeant species. These alterations were much larger for neutral membranes than for negatively charged ones. The dye increased the conductances mediated by positively charged permeant species and decreased those by negatively charged permeant species, suggesting that it produces a negative electrostatic potential on the membrane; it also altered the kinetics and the voltage dependencies of permeation by these charge carriers. The magnitudes of dye-mediated conductance changes were much larger for positively charged permeants than for negatively charged ones; also, changes in ionic strength altered these dye effects in opposite directions from those predicted by the Stern equation, and the dependence of the conductance alteration on dye concentration was steeper than that predicted by this equation. Finally, only very small changes in liposome zeta potentials were induced by the dye. Calculations show that a large fraction of these effects can be accounted for by the dipole potential produced by merocyanine at the membrane surface, but that additional effects of the dye must be postulated as well.

Coloring Agents↗
PubMed1981

Na+-dependent transport of tricarboxylic acid cycle intermediates by renal brush border membranes. Effects on fluorescence of a potential-sensitive cyanine dye.

The effect of the transport of tricarboxylic acid cycle intermediates on the membrane potential of renal brush border vesicles was studied using fluorescence of the cyanine dye, 3,3'-dipropylthiadicarbocyanine iodide. The behavior of the dye in the preparation was established with valinomycin-induced K+-diffusion potentials; increases in fluorescence were associated with depolarizing conditions. Addition of 1 mm succinate or citrate to membrane/dye suspensions produced transient increases in fluorescence, indicative of a depolarizing event(s) associated with the transport of these substrates. The transient response in fluorescence was Na+ dependent, of greater magnitude under Na+-gradient as compared to Na+-equilibrium conditions, and was a saturable function of substrate concentration. The specificity of the fluorescence response was identical to that obtained from studies of the competitive inhibition of succinate transport by tricarboxylic acid cycle intermediates and analogs We conclude that the major tricarboxylic acid cycle intermediates are transported via a common Na+-dependent transport system in renal brush border membranes.

Animals↗
PubMed1980

A model for the effects of potential and external K+ concentration on the Cs+ blocking of inward rectification.

Inward rectification in starfish egg cell membranes is blocked by external Cs+, the degree of blocking being an increasing function of hyperpolarizing potentials and of the external concentration of K+. While the effect of K+ and the particular functional dependence of blocking on potential are difficult to reconcile with a one-ion pore model, both features can be accounted for assuming that the pore has at least two sites and tat a considerable fraction of the blocked pores is simultaneously occupied by a Cs+ ion in the inner site and by a K+ in the outer one.

Animals↗
PubMed1980

Interactions of voltage-sensing dyes with membranes. I. Steady-state permeability behaviors induced by cyanine dyes.

The effects of a series of thiadicarbocyanine dyes, diSCn(5), in altering the electrical properties of lipid bilayer membranes have been studied as a function of the membrane's intrinsic surface-charge density, the aqueous ionic strength, and the length (n) of the hydrocarbon side chains on the dye. Zero-current conductances, transmembrane potentials, and conductance-voltage relationships induced by these dyes were measured. All dyes studied altered membrane permeability properties; however these alterations were much larger at lower (e.g. 10(-3) M) than at higher (e.g. 10(-1) M) ionic strengths. The data suggest that such perturbations would not be troublesome for most biological preparations in which these dyes have been studied. The mechanisms by which these dyes alter membrane permeabilities vary in going from short-chained to long-chained dyes, the former forming voltage-gated, ion-permeant pores and the latter acting predominantly as anion carriers (forming 2:1 dye-anion complexes). In the case of diSC3(5), the predominant mechanism of altering membrane permeabilities changes in going from neutral to negatively charged membranes and also depends upon aqueous ionic strength and dye concentration.

Carbocyanines↗
PubMed1980

Interactions of voltage-sensing dyes with membranes. II. Spectrophotometric and electrical correlates of cyanine-dye adsorption to membranes.

The adsorption to bilayer membranes of the thiadicarbocyanine dyes, diSCn(5), has been studied as a function of the membrane's surface-charge density, the aqueous ionic strength, and the length (n) of the hydrocarbon side chain of the dye. "Probe" measurements in planar bilayers, microelectrophoresis of liposomes, and measurement of changes in dye absorbance and fluorescence in liposomes were used to study dye adsorption to membranes. These measurements indicated that the membrane:water partition coefficient for the dye monomer increases with the length of the hydrocarbon side chain. However, the formation of large aggregates in the aqueous phase also increases with increasing chain length and ionic strength so that the actual dye adsorbing to the membrane goes through a maximum at high but not at low ionic strengths. More dye adsorbs to negatively charged than neutral membranes. Membrane-bound dye spectra were easily resolved in negatively charged liposomes where it was observed that these dyes could exist as monomers, dimers, and large aggregates. For diSC1(5) a spectral peak was observed at low but not high ionic strengths (i.e. the conditions in which this dye appears to form voltage-gated channels) corresponding to small aggregates which appeared to adsorb to the membrane. Finally, the adsorption of these dyes to membranes results in more positive electrostatic potentials composed primarily of dye-induced "boundary" potentials and somewhat less of "double-layer" potentials.

Carbocyanines↗
PubMed1978

A model for anomalous rectification: electrochemical-potential-dependent gating of membrane channels.

A model is presented for "anomalous rectification" based upon electrical measurements on the egg cell membrane of the starfish. The objective is to postulate a plausible molecular mechanism which yields an expression for the conductance similar to that deduced empirically by Hagiwara and Takahashi (1974), i.e.,: formula: (see text), where B, deltaVh and v are constant, cK is the external K+ concentration, and deltaV (= V - V0) is the displacement of the membrane potential from its resting value. It is shown that a similar dependence of the conductance on deltaV is expected for a particular class of models in which the K+ ions are also implicated in "gating". To give a specific example, we consider the case in which the formation of ion-permeable pores requires a voltage-induced orientation of membrane-bound, electrically-charged groups and subsequent complexation of these groups with the external cations. Furthermore, the proportionality between GK and CK1/2, when the internal K+ concentration is constant, is accounted for by conventional descriptions of the ionic fluxes using Eyring's rate reaction theory. In terms of the present model, B and deltaVh are explicit functions of the internal K+ concentrations and are thus constant only as long as this is unvaried. The particular value of v required to fit the data (v congruent to 8.4 mV) is rationalized by the assumption that each of the orientable groups carries three negative elementary charges. In addition, the predictions of the present model are compared with those deduced from an alternative viewpoint, which is related to Armstrong's "blocking particle hypothesis", in that the probability for opening and closing of the pore is assumed to depend on whether the pore is occupied or empty. Differences and similarities between the two models, as well as ways to discriminate between them, are discussed.

Animals↗
PubMed1977

Anomalous permeabilities of the egg cell membrane of a starfish in K+-Tl+ mixtures.

The electrical properties of "inward" rectifying egg cell membranes of the starfish mediastera aequalis have been studied in the presence of K(+)-Tl(+) mixtures. When the ratio of the external concentrations of these ions is changed while their sum is kept constant, both the conductance and the zero-current membrane potential go through a minimum, showing clear discrepancies from theoretical results based on conventional electrodiffusion models (E.g., Goldman's equation). By contrast, when the ration of the two concentrations is fixed and their sum varied, the potential follows an ideal Nernst slope, consistent with Goldman's equation. The membrane conductance which, according to previous studies on similar membranes, is to be viewed as a function of the displacement of the membrane potential from its resting value deltaV, shows marked differences between the cases in which K(+) or Tl(+) are the predominant ions: when K(+) is the predominant permeant ion in solution, the addition of small amounts of Tl(+) inhibits the current, while corresponding blocking effects of K(+) on the current are not observed when Tl(+) is the predominant permeant ion. Also, the time course of the conductance during voltage clamp is different in the two cases, being much faster in Tl(+) than in K(+) solution for comparable values of deltaV. Most of the above features are accounted for by a model in which it is assumed that the ionic channels have external binding sites for cations and that their permeability properties depend on the species of the cation bound (K(+)or Tl(+) in the present experiments).

Animals↗
PubMed1976

Influence of molecular variations of ionophore and lipid on the selective ion permeability of membranes: I. Tetranactin and the methylation of nonactin-type carriers.

The manner in which molecular structure of the carrier and the lipid composition of the membrane modulate the membrane selectivity among monovalent cations has been investigated for nonactin, trinactin, and tetranactin, which differ only in their degrees of methylation, and for membranes made of two lipids, phosphatidyl ethanolamine and glyceryl dioleate, in which "equilibrium" and "kinetic" aspects of permeation, respectively, are emphasized. Bilayer permeability ratios for Li, Na, K, Rb, Cs, Tl,and NH4 have been characterized and resolved into "equilibrium" and "kinetic" components using a model for carrier-mediated membrane transport which includes both a trapezoid energy barrier for translocation of the complex across the membrane interior and a potential-dependence of the loading and unloading of ions at the membrane-solution interfaces. The bilayer permeability properties due to tetranactin have been characterized in each of these lipids and found not only to be regular but to be systematically related to those of the less methylated homologues, trinactin and nonactin. This analysis has led to the following conclusions: (1) The change in lipid composition alters the relative contributions of "kinetic" vs. "equilibrium" components to the observed carrier-mediated selectivity. (2) Increased methylation of the carrier increases the contribution of the "kinetic" component to the selectivity relative to that of the "equilibrium" component and additionally alters the "equilibrium component sufficiently that an inversion of Cs--Na selectivity occurs between trinactin and tetranactin. (3) For all ions and carriers examined, the "reaction plane" for ion-carrier complexation and the width for the "diffusion barrier can be represented by the same two parameters, independent of the ion or carrier, so that in all cases the complexation reaction senses 10% of the applied potential and the plateau of the "diffusion barrier" extends across 70% of the membrane interior.

Anti-Bacterial Agents↗
PubMed1971

Freezing and melting of lipid bilayers and the mode of action of nonactin, valinomycin, and gramicidin.

An abrupt loss of effectiveness of the presumed carriers, nonactin and valinomycin, in mediating ion conductance occurred at the same temperature as the membrane fluidity, judged visually, was lost. By contrast, the effects of the presumed channel-former, gramicidin, were the same on solid and liquid membranes. Taken together, these findings imply that freezing the membrane primarily reduces the mobility of these antibiotics with little effect on their solubility.

Anti-Bacterial Agents↗