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Cation-dependent segregation phenomena and phase behavior in model membrane systems containing phosphatidylserine: influence of cholesterol and acyl chain composition.

The addition of Ca2+ to model membrane systems containing phosphatidylserine (PS) can have remarkable effects on the distribution of PS and the overall polymorphic phase [bilayer or hexagonal (HII)] assumed by the lipid mixture. In this study, we examine the influence of Ca2+ on lipid mixtures composed of well-defined (synthetic) species of PS, phosphatidylethanolamine (PE), and phosphatidylcholine (PC) in the presence and absence of cholesterol by employing 31P and 2H NMR, freeze-fracture, and X-ray techniques. It is shown that whereas Ca2+ can segregate PS into crystalline cochleate domains in equimolar mixtures of dioleoyl-PE and dioleoyl-PS (DOPS), such effects are not observed for mixtures containing more unsaturated (dilinoleoyl) species of PS. The addition of cholesterol to these PE-PS systems inhibits Ca2+-induced segregation of DOPS and facilitates Ca2+-triggered hexagonal (HII) phase formation for both the PE and the PS components. In contrast, in equimolar mixtures of DOPS with dioleoyl-PC, Ca2+-induced segregation of phospholipid is not affected by the presence of up to 33 mol % cholesterol. These and related effects suggest that, in multicomponent biomembrane systems containing both PE and cholesterol, phase segregation of PS by Ca2+ may not be readily achievable. These results are discussed with regard to the reliability of 31P NMR phase identifications of phospholipid structure in model and biological membranes and demonstrate that in mixed lipid systems the influence of divalent cations on lipid distribution and structure can be exquisitely sensitive to details of the local lipid composition.(ABSTRACT TRUNCATED AT 250 WORDS)

Cations↗

Divalent cation induced fusion and lipid lateral segregation in phosphatidylcholine-phosphatidic acid vesicles.

The interactions of unilamellar vesicles containing phosphatidylcholine (PC) and phosphatidic acid (PA) in the presence of calcium and magnesium were examined by fluorometric assays of vesicle lipid mixing, contents mixing, and contents leakage and by spray-freezing freeze-fracture electron microscopy. These results were correlated with calorimetric and fluorometric measurements of divalent cation induced lateral segregation of lipids in these vesicles under comparable conditions. PA-PC vesicles in the presence of calcium show a rapid but limited intermixing of vesicle lipids and contents, the extent of which increases as the vesicle size decreases or the PA content increases. Calcium produces massive aggregation and efficient mixing of the contents of vesicles containing high proportions of dioleoyl-PA or egg PA, but vesicle coalescence in the latter case is followed rapidly by vesicle collapse and massive leakage of contents. The effects of magnesium are similar for vesicles of very high PA content. However, in the presence of magnesium, vesicles containing lower amounts of PA exhibit "hemifusion", a mode of interaction in which vesicles aggregate and mix approximately 50% of their lipids, apparently representing the lipids of the outer monolayer of each vesicle, without significant mixing of vesicle contents or collapse of the vesicles. Fluorometric measurements of lipid lateral segregation demonstrate that lateral redistribution of lipids in PA-PC vesicles begins at submillimolar concentrations of divalent cations and shows no abrupt change at the "threshold" divalent cation concentration, above which coalescence of vesicles is observed. By correlating calorimetric and fluorometric measurements of lipid lateral segregation and mixing of vesicle components, we can demonstrate that lipid segregation is at least strongly correlated with calcium-promoted coalescence of PA-PC vesicles and is essential to the magnesium-promoted interactions of vesicles of low PA contents.

Cations, Divalent↗

Nano-segregated polymeric film exhibiting high ionic conductivities.

Nanostructures can be used for the fabrication of highly functional materials transporting ions and charges. We demonstrate a new design strategy for polymeric higher ion-conductors. Phase-segregated layers of alternating mobile tetra(ethylene oxide)s (TEOs) and rigid aromatic cores where the TEO moieties are grafted from aromatic layers have been shown to be efficient to transport lithium triflate. Such segregated structures at the nanometer scale (nano-segregated structures) were prepared by in-situ photopolymerization of an aligned methacrylate liquid crystalline monomer comprising a terphenyl rigid rod mesogen having a TEO terminal chain. The ion-conductive TEO moiety remains in the highly mobile state even after polymerization, which is indicated by its low glass transition temperature (-45 degrees C). This nanostructured film exhibits an ionic conductivity parallel to the layer of 10(-3) S cm(-1) at room temperature. The highest ionic conductivity is in the level of 10(-2) S cm(-1) observed at 150 degrees C. The anisotropic ionic conductivities have been observed for the nano-segregated film.

Journal Article↗

Nanophase segregation and water dynamics in the dendrion diblock copolymer formed from the Fréchet polyaryl ethereal dendrimer and linear PTFE.

We propose a new material consisting of a dendrion copolymer formed from (a) a water-soluble dendritic polymer and (b) a hydrophobic backbone. Using molecular dynamics simulations techniques, we determine the structure and dynamics of the dendrion formed by second-generation Fréchet polyaryl ethereal dendrimer as the hydrophilic component and linear polytetrafluoroethylene (PTFE) as the hydrophobic polymer, with 5 and 10 wt % of water. We find that this material produces a well-developed nanoscale structure in which water forms a continuous nanophase, making this new family of compounds promising candidates for applications in fuel cell membranes. We find that the water molecules are incorporated into the dendrimer block of the copolymer to form a nanophase-segregated structure. The well-developed nanophase-segregated structures rendered by this material have characteristic dimensions of segregation ( approximately 30 Angstrom) and dendrimer conformational properties that are independent of water content. Calculations of water dynamics and proton transport in these nanophase-segregated structures indicate that the dendrion copolymer membrane with 10 wt % of water content has a water structure and transport properties equivalent to that of the hydrated Nafion membrane with 20 wt % of water content.

Dendrimers↗

Associative and segregative phase separations of gelatin/kappa-carrageenan aqueous mixtures.

The effects of ionic strength, temperature, and pH on the phase separation behavior of type B pigskin gelatin/sodium-type kappa-carrageenan aqueous mixtures were investigated. Depending on the different combinations of temperature and sodium chloride (NaCl) concentration, the mixtures showed compatible, associative, and segregative phase separation behaviors. Additionally, a coexistence of associative and segregative (associative-co-segregative) phase separations was expected at low temperature and low NaCl concentration. These different phase separation events were observed using confocal scanning laser microscopy. Moreover, it was found that the segregative phase separation when alone is induced by the ordering of kappa-carrageenan chains, while that in the coexistence region is induced by the ordering of gelatin chains. pH had a significant effect on the associative phase separation, resulting in morphologies changing from compatible solution to liquid coacervate and further to solid precipitate with decreasing pH. These were attributed to the dramatic changes of the charge density of amphoteric gelatin during the pH decrease.

Carrageenan↗

Effect of rhythmic grouping on stream segregation.

Three experiments investigated the role of temporal grouping on auditory stream segregation. For sounds that formed frequency streams (e.g., 400 Hz, 500 Hz, 1600 Hz, and 2000 Hz), the effect of rhythm was minimal. Temporal grouping did not affect judgments of stream segregation and did not affect difficulty of sequence identification. In contrast, for sounds that tended to form one coherent sequence (e.g., 750 Hz, 1500 Hz, 3000 Hz, and 6000 Hz), temporal grouping affected judgments of stream segregation as well as difficulty of identification. The temporal grouping could space the three lower or three higher pitch tones equally in time and this induced isochronous stream segregation. Subjects could not interleave the resulting streams, and identification became far more difficult. The role of rhythmic grouping is therefore contextual, depending on the relationships between the elements as well as the order of the elements.

Auditory Perception↗

Mediation of meiotic and early mitotic chromosome segregation in Drosophila by a protein related to kinesin.

Contrary to the traditional view that microtubules pull chromosomes polewards during the anaphase stage of meiotic and mitotic cell divisions, new evidence suggests that the chromosome movements are driven by a motor located at the kinetochore. The process of chromosome segregation involves proper arrangement of kinetochores for spindle attachment, followed by spindle attachment and chromosome movement. Mechanisms in Drosophila for chromosome segregation in meiosis differ in males and females, implying the action of different gene products in the two sexes. A product encoded at the claret locus in Drosophila is required for normal chromosome segregation in meiosis in females and in early mitotic divisions of the embryo. Here we show that the predicted amino-acid sequence of this product is related to the heavy chain of kinesin. The conserved region corresponds to the kinesin motor domain and includes the ATP-binding site and a region that can bind microtubules. A second region contains a leucine repeat motif which may mediate protein-subunit interactions necessary for attachment of chromosomes to the spindle. The mutant phenotype of chromosome nondisjunction and loss, and its similarity to the kinesin ATP-binding domain, suggest that the product encoded at claret not only stabilizes chromosome attachments to the spindle, but may also be a motor that drives chromosome segregation in female meiosis.

Adenosine Triphosphatases↗

Asymmetric segregation of the homeodomain protein Prospero during Drosophila development.

Asymmetric divisions that produce two distinct cells play fundamental roles in generating different cell types during development. In the Drosophila central nervous system, neural stem cells called neuroblasts divide unequally into another neuroblast and a ganglion mother cell which is subsequently cleaved into neurons. Correct gene expression of ganglion mother cells requires the transcription factor Prospero. Here we demonstrate the asymmetric segregation of Prospero on neuroblast division. Prospero synthesized in neuroblasts is retained in the cytoplasm and at mitosis is exclusively partitioned to ganglion mother cells, in which it is translocated to the nucleus. Differential segregation of Prospero was also found in the endoderm. We have identified a region in Prospero that is responsible for this event. The region shares a common motif with Numb, which also shows unequal segregation. We propose that asymmetric segregation of transcription factors is an intrinsic mechanism for establishing asymmetry in gene expression between sibling cells.

Amino Acid Sequence↗

Methods of segregation analysis for animal breeding data: a comparison of power.

Maximum likelihood segregation analysis provides potentially the most powerful method for the detection of segregating major genes. Segregation analysis requires the comparison of the likelihood of the data under the combined model (allowing both polygenic and major gene genetic variation) with the likelihood of the data under the polygenic model (allowing only polygenic genetic variation). In this study three approximations to the combined model likelihood were compared using simulated data, both with and without a segregating major gene, containing observations on paternal half-sibs. The use of Hermite integration to replace the integration in the combined model likelihood provided the most powerful test for a major gene. Two approximations, based on extensions of linear-mixed-model theory and estimating transmitting abilities for sires, were also considered. These approximations were less powerful than the use of Hermite integration, although the approximation estimating a transmitting ability for each major genotype for the sires was an improvement over the approximation estimating a single transmitting ability. For each approximation the frequency of detection of a major gene depended on the proportion of the genetic variance explained by the simulated major gene and whether the major gene caused the distribution to be skewed.

Alleles↗

The bacterial segrosome: a dynamic nucleoprotein machine for DNA trafficking and segregation.

The genomes of unicellular and multicellular organisms must be partitioned equitably in coordination with cytokinesis to ensure faithful transmission of duplicated genetic material to daughter cells. Bacteria use sophisticated molecular mechanisms to guarantee accurate segregation of both plasmids and chromosomes at cell division. Plasmid segregation is most commonly mediated by a Walker-type ATPase and one of many DNA-binding proteins that assemble on a cis-acting centromere to form a nucleoprotein complex (the segrosome) that mediates intracellular plasmid transport. Bacterial chromosome segregation involves a multipartite strategy in which several discrete protein complexes potentially participate. Shedding light on the basis of genome segregation in bacteria could indicate new strategies aimed at combating pathogenic and antibiotic-resistant bacteria.

Amino Acid Sequence↗

Chromosomal segregation and fertility in Robertsonian chromosomal heterozygotes of the house musk shrew (Suncus murinus, Insectivora, Soricidae).

Crucial to our understanding of chromosomal variation and evolution in mammals are detailed studies of chromosomal heterozygotes, with analyses of chromosomal segregation and chromosome-derived infertility. We studied segregation and fertility in hybrids between karyotypic races of the house musk shrew Suncus murinus. These individuals were heterozygous for up to five Robertsonian fusions (Rbs) and an insertion of heterochromatin in an autosome. All variant chromosomes showed Mendelian segregation and all Rbs segregated independently of each other in the progeny of double heterozygotes. Litter size in single and even multiple Rb heterozygotes was no smaller than that in the less fertile parental strain. The effects of genetic background were more important in determining litter size than Rb heterozygosity for the shrews that we examined.

Animals↗

Quantification of boundary segregation in the analytical electron microscope

When studying equilibrium grain-boundary segregation using the small ( approximately 1 nm) electron probe of the scanning transmission electron microscope and X-ray energy dispersive spectroscopy, the assumptions made about the size and shape of the beam-specimen interaction volume may introduce errors in quantification of up to a factor of five. Comparisons between experimental segregation profiles and different theoretical models have shown that a Gaussian model for the electron distribution will provide the best description for the interaction volume. Calculations of minimum detectable segregation levels have shown that optimum sensitivity is not achieved in the thinnest samples or even with the smallest probe sizes. In addition, operating at 300 keV (rather than 100 keV) will halve the minimum detectable segregation level, assuming all other experimental conditions are equal. Rastering the electron probe over a fixed area while acquiring spectra improves the accuracy of quantification but at the price of reducing sensitivity by at least a factor of three.

Journal Article↗

Calcium transients accompany ooplasmic segregation in zebrafish embryos.

Through the injection of f-aequorin (a calcium-specific luminescent reporter), and the use of an imaging photon detector, transient localized elevations of free cytosolic calcium in the forming blastodisc (BD) and animal hemisphere cortex were visualized that correlated with ooplasmic segregation. The introduction of an appropriate concentration of the weak (KD = 1.5 micromol/L) calcium buffer 5,5'-dibromo-BAPTA results in the dissipation of these calcium domains, and inhibits cytoplasmic streaming and the subsequent formation of a BD at the animal pole. These inhibitory actions are dependent on the final cytosolic concentration of buffer within the egg: > or = 1.3 mmol/L blocks ooplasmic streaming; < 1.3 mmol/L eggs segregate normally. Injection of 5,5'-dimethyl-BAPTA (KD = 0.15 micromol/L) to a final concentration of 1.5 mmol/L as a control has no effect on ooplasmic streaming. These results suggest that localized domains of elevated free cytosolic calcium are essential for ooplasmic segregation in zebrafish. Furthermore, a hypothetical model is presented linking these calcium transients to the contraction of a cortically located actin microfilament network as a possible mechanism providing the driving force for segregation.

Aequorin↗

Convergence of segregated pheromonal pathways from the accessory olfactory bulb to the cortex in the mouse.

The accessory olfactory system mediates intraspecies pheromonal communication. Two subsets of spatially segregated vomeronasal sensory neurons, presumably handling functionally and structurally different sets of ligand molecules, can be distinguished. The two subsets of sensory neurons project their axons to segregated zones of the accessory olfactory bulb (AOB) and connect with zonally separated mitral/tufted (M/T) cells, suggesting that the accessory olfactory system is divided into two distinct pathways up to the level of the AOB. To examine whether the segregation is maintained at the accessory olfactory cortical (AOC) regions, we selectively tracer-labelled mitral/tufted cells located in the rostral, caudal or in both zones of the adult mouse AOB. The results demonstrate that the axonal projection patterns of rostral zone and caudal zone M/T cells were indistinguishable in the AOC regions. Mitral/tufted cell axons from either zone of the AOB covered the entire area of all four AOC regions: the bed nucleus of the accessory olfactory tract, the medial amygdaloid nucleus, the posteromedial cortical amygdaloid nucleus and the bed nucleus of the stria terminalis. Therefore, over the entire area of each AOC region, ensembles of cortical neurons receive input from both zonal subsets of M/T cells of the AOB. However, the present results do not rule out the possibility that individual cortical neurons sample information from M/T cells of a single zone. These results are consistent with the idea that the segregation of zonal pathways collapses in the AOC regions. Clusters of cortical neurons in each AOC region may combine information from both families of pheromone receptors and thus handle signals from structurally and functionally different categories of pheromone molecules.

Animals↗

Development of a microsatellite-based approach to co-segregation analysis of familial hypercholesterolaemic kindreds.

Co-segregation studies based on a selection of intragenic restriction fragment length polymorphisms of the low density lipoprotein receptor (LDLR) gene have been used extensively both for research and diagnostic studies of familial hypercholesterolaemia (FH) families, because direct mutation screening remains complex. Here we describe the development and application of a more efficient approach to co-segregation studies based on highly informative dinucleotide and tetranucleotide repeats flanking the LDLR gene. A series of microsatellites (D19S391, D19S394, D19S221 and D19S179) were selected for study on the basis of linkage analysis in the CEPH families using intragenic polymorphisms for a TA repeat (exon 18) in the LDLR gene, and earlier data for a Pvu II polymorphism (intron 15). A physical map of the region of chromosome 19 also contributed to this selection. One marker in particular, D19S394, sited 150 kilobases telomeric to the gene, was extremely useful, displaying 90% heterozygosity, robust PCR of tetranucleotide repeats without stutter bands, and no recombination with the LDLR gene (theta = 0, LOD 68). Use of this marker in the families of twenty-three FH probands from Hampshire demonstrated co-segregation of the hyperlipidaemia phenotype with the LDLR gene region, except in one family with defective apolipoprotein B-100, and a family turning out to display familial combined hyperlipidaemia. This approach should facilitate the search for any families where FH does not co-segregate with the LDLR gene, and will enhance the repertoire of molecular diagnostic tools available for FH.

Adolescent↗

Configurational bias Monte Carlo simulation of phase segregation in block copolymer networks.

Cross-linked block copolymers are used as adhesives in fiber-reinforced composite material manufactures for automotive applications. Good adhesion between the polymer matrix and fibers in the interphase region is required for the structural integrity of these materials. Experimental evidence indicates that superior adhesion is obtained when phase segregation occurs between the two matrix phase block copolymers. It is therefore desirable to predict the conditions under which phase segregation is expected to occur. Configurational bias Monte Carlo simulations of two-component, trifunctional block copolymer networks were carried out to investigate phase segregation in these materials. The effects of four principal parameters on phase segregation were examined: the weight fractions of the two components, the cross-link length, the connectivity of the network, and the ratio of the square-well interactions. The molecular simulation results confirmed trends observed in laboratory measurements.

Journal Article↗

Segregation induced instabilities of granular fronts.

Experimental investigation of granular flows containing particles of several sizes and moving down slopes shows that segregation of coarse-grained, irregularly shaped particles induces a fingering instability at the propagating front. The size-segregation mechanism involves percolation of small particles downward and a corresponding migration of large ones toward the flow surface. Large particles at the flow surface experience velocities that are greater than average so that they migrate forward and begin to collect at the flow front. In the case of dry cohesionless flows, the instability depends upon these large particles at the flow perimeter being more angular and thus more resistant to flow than the smaller rounder ones in the interior. A simple analytical model predicts the fingering instability when friction of the flow front is greater than that of the following flow. The presence of viscous liquid inhibits both size-segregation and the development of the instability. Fluidization of dry flows permits segregation of large particles to flow perimeters, thus increasing permeability and permitting a similar instability that owes its development to the dry frictional perimeter that surrounds a partly fluidized interior. (c) 1999 American Institute of Physics.

Journal Article↗

Monte Carlo simulations of segregation in Pt-Re catalyst nanoparticles.

We have investigated the segregation of Pt atoms to the surfaces of Pt-Re nanoparticles using the Monte Carlo method and modified embedded-atom method potentials that we have developed for Pt-Re alloys. The Pt(75)Re(25) nanoparticles (containing from 586 to 4,033 atoms) are assumed to have disordered fcc configurations and cubo-octahedral shapes (terminated by [111] and [100] facets), while the Pt(50)Re(50) and Pt(25)Re(75) nanoparticles (containing from 587 to 4,061 atoms) are assumed to have disordered hcp configurations and truncated hexagonal bipyramidal shapes (terminated by [0001] and [1011] facets). We predict that due to the segregation process the equilibrium Pt-Re nanoparticles would achieve a core-shell structure, with a Pt-enriched shell surrounding a Pt-deficient core. For fcc cubo-octahedral Pt(75)Re(25) nanoparticles, the shells consist of almost 100 at. % of Pt atoms. Even in the shells of hcp truncated hexagonal bipyramidal Pt(50)Re(50) nanoparticles, the concentrations of Pt atoms exceed 85 at. % (35 at. % higher than the overall concentration of Pt atoms in these nanoparticles). Most prominently, all Pt atoms will segregate to the surfaces in the hcp truncated hexagonal bipyramidal Pt(25)Re(75) nanoparticles containing less than 1000 atoms. We also find that the Pt atoms segregate preferentially to the vertex sites, less to edge sites, and least to facet sites on the shell of Pt-Re nanoparticles.

Journal Article↗