The role of passive calcium influx through the cell membrane in galvanotaxis

Passive calcium influx is one of the theories to explain the cathodal galvanotaxis of cells that utilize the electric field to guide their motion. When exposed to an electric field, the intracellular fluid becomes polarized, leading to positive charge accumulation on the cathodal side and negative charge accumulation on the anodal side. The negative charge on the anodal side attracts extracellular calcium ions, increasing the anodal calcium concentration, which is supposed to decrease the mobile properties of this side. Unfortunately, this model does not capture the Ca²⁺ dynamics after its presentation to the intracellular fluid. The ions cannot permanently accumulate on the anodal side because that would build a potential drop across the cytoplasm leading to an ionic current, which would carry positive ions (not only Ca²⁺) from the anodal to the cathodal part through the cytoplasm. If the cytoplasmic conductance for Ca²⁺ is low enough compared to the membrane conductance, the theory could correctly predict the actual behavior. If the ions move through the cytoplasm at a faster rate, compensating for the passive influx, this theory may fail. This paper contains a discussion of the regimes of validity for this theory.     

Energy parameters of Acetabularia cell membrane]

The energy parameters of the mechanism involved in the action of Acetabularia cell membrane were studied. The resistance of the membrane is found to be near 3 kohm per sq. cm when resting and at the peak of excitation it is much lower (100 ohm/cm(2). The power of the mechanism regulating the action of the membrane during the peak. The total power of the membrane system of Acetabularia is 8 microwatt per sp. cm. reaches 6.5 microwatt/cm(2). The energy consumed by the regulatory mechanism during the action potential is minus 290 plus or minus 20 erg/cm(2) and the total energy dissipation is 20 erg/cm(2). The values of the energy consumed by the cell during the action potential are compared with its energy resource.     

Changes in passive electric parameters of human erythrocyte membrane during hyperthermia: role of spectrin phosphorylation.

In prefixed by 1 mmol/l OsO4 human erythrocytes, the discocyte shape was preserved upon heating to temperatures which include the denaturation temperature of the main peripheral protein spectrin. Nevertheless, the suspension of fixed cells displayed threshold decrease in its capacitance and resistance at the temperature range where spectrin denaturates. The same changes were established using intact cells and their resealed ghosts. For packed cells (ghosts), the capacitance and resistance decreased about 17% (31%) and 30% (19%). These data indicate a decrease in the beta dispersion of erythrocyte membrane associated, according to a previous study (Ivanov 1997), with the heat denaturation of spectrin at 49.5 degrees C. The amplitude of the 49.5 degrees C decrease in beta dispersion was reversibly reduced in intact erythrocytes and white ghosts following reversible decrease in the phosphorylation of their membrane proteins. It was fully eliminated in ghosts following their resealing with alkaline phosphatase (0.1 mg/ml) which dephosphorylated membrane proteins. These findings are discussed in relation to similar changes found in normal and tumour tissues and cells during hyperthermia.     

The ultrastructure of the oyster brown cell,a cell with a fenestrated plasma membrane

Summary The oyster brown cell, a connective tissue cell of uncertain function and affinity, was characterized in the electron microscope by (1) the presence of large cytoplasmic granules, (2) fenestrations of the plasma membrane, and (3) an extensive tubular network originating in, or emptying into, the plasma membrane fenestrations. The brown cell did not appear to be a cell involved in glycogen storage or in the manufacture of exportable protein. The extensive tubular network and the membrane slits suggested that the brown cell may have been involved in the processing of biological fluids.This work was supported in part by Public Health Service Contract No. 5 To 1 ES00038-02, Health Sciences Advancement Award No. RR06138, and Tumor Biology Training Grant, NIH CA 05245.We wish to thank Miss Grete Nilsen for her expert technical assistance and Mr. Bob Munn for his help in the use of the electron microscope and for proof reading our MS. Our appreciation is also extended to Dr. J. Luft, Dr. A. K. Sparks, Miss P. Phelps, Mr. M. DeVault, and to the personnel of the Johnson Oyster Company, Inverness.     

The effect of cell excision and microelectrode perforation on membrane resistance measurements of Nitella translucens.

The effect of cell excision and microelectrode perforation on the membrane resistance measurements of Nitella translucens was determined by direct experiment. From the results it is concluded that perforation has no effect on cells as short as 1 cm. Current leakage though the node of an excised cell has however to be given some consideration. The method used for determining the resistance recovery to insertion has a wide application and its simplicity will encourage its use in other biological systems.     

Modulation of P-glycoprotein-mediated multidrug resistance by acceleration of passive drug permeation across the plasma membrane

The drug concentration inside multidrug-resistant cells is the outcome of competition between the active export of drugs by drug efflux pumps, such as P-glycoprotein (Pgp), and the passive permeation of drugs across the plasma membrane. Thus, reversal of multidrug resistance (MDR) can occur either by inhibition of the efflux pumps or by acceleration of the drug permeation. Among the hundreds of established modulators of Pgp-mediated MDR, there are numerous surface-active agents potentially capable of accelerating drug transbilayer movement. The aim of the present study was to determine whether these agents modulate MDR by interfering with the active efflux of drugs or by allowing for accelerated passive permeation across the plasma membrane. Whereas Pluronic P85, Tween-20, Triton X-100 and Cremophor EL modulated MDR by inhibition of Pgp-mediated efflux, with no appreciable effect on transbilayer movement of drugs, the anesthetics chloroform, benzyl alcohol, diethyl ether and propofol modulated MDR by accelerating transbilayer movement of drugs, with no concomitant inhibition of Pgp-mediated efflux. At higher concentrations than those required for modulation, the anesthetics accelerated the passive permeation to such an extent that it was not possible to estimate Pgp activity. The capacity of the surface-active agents to accelerate passive drug transbilayer movement was not correlated with their fluidizing characteristics, measured as fluorescence anisotropy of 1-(4-trimethylammonium)-6-phenyl-1,3,5-hexatriene. This compound is located among the headgroups of the phospholipids and does not reflect the fluidity in the lipid core of the membranes where the limiting step of drug permeation, namely drug flip-flop, occurs.     

The red cell membrane and its cytoskeleton.

Gel-filtration (Sephadex G-75) analysis of hepatic cytosol reveals both qualitative and quantitative sex differences in oestrogen-binding proteins. The elution profile of [³H]oestradiol-labelled cytosol shows four species of oestrogen-binding proteins (peaks I, II, IV and V) common to both sexes. The amount of [³H]oestradiol binding in peak I is equivalent in both males and females and corresponds quantitatively to the specific oestrogen receptor. The amount of binding in the remaining three peaks is greater in males than females. In addition, an oestrogen-binding protein (peak III) is present that is unique to male cytosol. Proteinase-inhibition studies demonstrate that the observed multiplicity of oestrogen-binding proteins is not an artefact of proteolytic breakdown. Sex differences in oestrogen-binding proteins are absent in immature male and female animals; the oestrogen-binding protein profile in immature rats resembles that of an adult female. Gonadectomy of adult animals does not affect the oestrogen-binding-protein profile. In contrast, neonatal (day 1) castration results in partial feminization of the characteristic oestrogen-binding protein profile seen in the adult male; the appearance of Peak III is suppressed and marked decreases in the amount of oestradiol binding occurs in the remaining peaks. Hypophysectomy of adult animals results in near abolishment of the observed sex differences; the male oestrogen-binding protein profile is partially feminized and the female profile is partially masculinized, as characterized by the appearance of [³H]oestradiol binding in the region of peak III and increased amounts of binding in peaks IV and V. The present studies demonstrate a multiplicity of oestrogen-binding proteins in liver cytosol and raise the possibility that the presence of some of these proteins may be imprinted at birth through the hypothalamic–pituitary axis, by a mechanism requiring neonatal androgen exposure.     

Daptomycin resistance in enterococci is associated with distinct alterations of cell membrane phospholipid content

Background

The lipopeptide antibiotic, daptomycin (DAP) interacts with the bacterial cell membrane (CM). Development of DAP resistance during therapy in a clinical strain of Enterococcus faecalis was associated with mutations in genes encoding enzymes involved in cell envelope homeostasis and phospholipid metabolism. Here we characterized changes in CM phospholipid profiles associated with development of DAP resistance in clinical enterococcal strains.

Methodology

Using two clinical strain-pairs of DAP-susceptible and DAP-resistant E. faecalis (S613 vs. R712) and E. faecium (S447 vs. R446) recovered before and after DAP therapy, we compared four distinct CM profiles: phospholipid content, fatty acid composition, membrane fluidity and capacity to be permeabilized and/or depolarized by DAP. Additionally, we characterized the cell envelope of the E. faecium strain-pair by transmission electron microscopy and determined the relative cell surface charge of both strain-pairs.

Principal Findings

Both E. faecalis and E. faecium mainly contained four major CM PLs: phosphatidylglycerol (PG), cardiolipin, lysyl-phosphatidylglycerol (L-PG) and glycerolphospho-diglycodiacylglycerol (GP-DGDAG). In addition, E. faecalis CMs (but not E. faecium) also contained: i) phosphatidic acid; and ii) two other unknown species of amino-containing PLs. Development of DAP resistance in both enterococcal species was associated with a significant decrease in CM fluidity and PG content, with a concomitant increase in GP-DGDAG. The strain-pairs did not differ in their outer CM translocation (flipping) of amino-containing PLs. Fatty acid content did not change in the E. faecalis strain-pair, whereas a significant decrease in unsaturated fatty acids was observed in the DAP-resistant E. faecium isolate R446 (vs S447). Resistance to DAP in E. faecium was associated with distinct structural alterations of the cell envelope and cell wall thickening, as well as a decreased ability of DAP to depolarize and permeabilize the CM.

Conclusion

Distinct alterations in PL content and fatty acid composition are associated with development of enterococcal DAP resistance.     

KCl leakage from microelectrodes and its impact on the membrane parameters of a nonexcitable cell

The Journal of Membrane Biology - Microcapillary electrodes filled with a variety of salt solutions, including 1m KCl, have been used to measure the membrane potentials and resistances of spherical...     

The chemical composition of wool. XV. The cell membrane couplex.

The cell membrane complex of wool has been examined by electron microscopy of stained cross sections after immersion of the wool in formic acid. The cell membrane complex of the cortex is considerably modified by the treatment, but that of the cuticle appears unchanged. Resistant membranes from cuticle cells, cortical cells and wool have been prepared by treatment with performic acid-ammonia. Amino acid analyses show that the resistant membranes from the cuticle contain citrulline but those from cortical cells do not. It is concluded that the cell membrane complex of the cuticle differs from that of the cortex. Because of the high lysine content of the resistant membranes, their resistance to chemical attack, the hydrophobicity of epicuticle and the observation of a small amount of epsilon-(gamma-glutamyl)lysine, it is postulated that the resistant membranes may contain an appreciable amount of epsilon-(gamma-glutamyl)lysine cross links.     

Unit membrane parameters of electrically syncytial tissues.

A change in the holding voltage, exposure to channel-blocking agents, and similar interventions will induce changes in the membrane properties of electrically syncytial tissues. The altered membrane characteristics will produce changes in the input resistance (RIN) and the phase angle (phi) of the complex admittance of the whole preparation. Exact geometry-independent formulas are derived that give the intervention-induced changes in the membrane capacitance and conductance in terms of the measured changes in RIN and phi. The formulas automatically account for the effects of extracellular resistance in tissues such as skeletal muscle fibers, cardiac Purkinje fibers, and small cardiac "aggregates." The size, shape, and resistance of the extracellular space may be arbitrary and need not be measured. The surface (invaginated) membranes, which face the bath (extracellular space), are assumed to be characterized by an RC circuit with specific capacity Cme (Cmi) and specific conductivity gme (gmi). It is assumed that the intracellular voltage gradient between the electrodes and the membranes is negligible or reliably calculable. The intervention is assumed to leave the geometry and resistivity of the extracellular space unchanged. Under these circumstances the intervention-induced changes in Cme, Cmi, gme, and gmi are determined exactly in terms of the corresponding changes in RIN and certain frequency domain integrals over phi. The technique is illustrated by synthetic data for RIN and phi generated by the "disk" model of a skeletal muscle fiber in which Cme and Cmi depend upon holding voltage. The corresponding voltage dependence of RIN and phi is successfully "inverted" to expose the underlying voltage dependence of Cme and Cmi. These computations suggest that the formulas for Cme and Cmi will be useful in realistic situations, since they are not too sensitive to experimental error in the data for RIN and phi. This method makes it possible to detect voltage-dependent capacity changes due to unit membrane processes (e.g., charge movement) as long as the intrinsic time constant of that process is very small (e.g., less than 1/30 ms). As a second example I consider a disk model that is exposed to increasing concentrations of a channel-blocking agent. The drug dependence of RIN and phi is used to calculate the drug dependence of the total membrane conductivity (the sum of gme and gmi, weighted by the areas of surface and invaginated membranes, respectively).     

Association of ferritin with liver cell membrane fractions.

Following intraperitoneal injection of rats with a large dose of ferric ammonium citrate containing ⁵⁹Fe, some 35 to 50% of the dose was deposited in the liver within the first 1 to 4 h. Almost all of the deposited iron could be precipitated with a ferritin antiserum from a homogenate of liver heated to 75 °C, but only half of this was precipitable when the unheated homogenate was treated with antiserum. The remainder of the ferritin iron was made available to the antiserum by treatment with deoxycholate, and was therefore presumed to be associated with membranous components of the cell.Subcellular fractionation of the liver following administration of ⁵⁹Fe-labeled ferric ammonium citrate showed that most of the radioactivity deposited within the first 4 h was equally divided between the cell sap and a light microsome (membrane-rich) fraction. Ferritin in this latter fraction was made available to antibody following deoxycholate treatment. The liver microsome fraction of the young rat contains little unavailable ferritin, but with aging there is an accumulation of ferritin in the microsomal fraction which is unavailable to antibody until the membrane is removed.It is suggested that at least part of the injected iron salt is taken up by pinocytotic vesicles and transferred to ferritin within this fraction, possibly followed by release of some of this ferritin into the cell sap.     

The plasmalemma redox system of a fresh-water alga and membrane electrical parameters

In order to learn more details about the plasmalemma redox system in the alga Hydrodictyon reticulatum the electrophysiological characteristics of the plasma membrane as influenced by artificial electron acceptors were estimated. Ferricyanide anion as well as TTF+ depolarized the membrane potential, the effect being more marked in the dark than in the light. In the presence of ferricyanide the membrane resistance in the light was decreased by about 29% on the average, in the dark it remained unchanged. On the other hand, TTF+ brought about a large increase in membrane resistance, notwithstanding the light conditions. The results are discussed in view of the impairing influence of both electron acceptors on the active inflow of chloride ions (Rybová, R. et al. (1990) Bot. Acta 103, 404-407). The electrogenic outflow of electrons appears to make the largest contribution to the membrane depolarization. The inhibition of a pumping mechanism coupling the active uptake of Cl- in some way to the transmembrane electron flow is envisaged as a plausible explanation for membrane resistance increase by TTF+.