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Microdomains
Kinds of Microdomains Selected AbstractsWhen is high-Ca2+ microdomain required for mitochondrial Ca2+ uptake?,ACTA PHYSIOLOGICA, Issue 1 2009A. Spät Abstract Ca2+ release from IP3 -sensitive stores in the endoplasmic reticulum (ER) induced by Ca2+ -mobilizing agonists generates high-Ca2+ microdomains between ER vesicles and neighbouring mitochondria. Here we present a model that describes when such microdomains are required and when submicromolar [Ca2+] is sufficient for mitochondrial Ca2+ uptake. Mitochondrial Ca2+ uptake rate in angiotensin II-stimulated H295R adrenocortical cells correlates with the proximity between ER vesicles and the mitochondrion, reflecting the uptake promoting effect of high-Ca2+ peri-mitochondrial microdomains. Silencing or inhibition of p38 mitogen-activated protein kinase (MAPK) or inhibition of the novel isoforms of protein kinase C enhances mitochondrial Ca2+ uptake and abolishes the positive correlation between Ca2+ uptake and ER-mitochondrion proximity. Inhibition of protein phosphatases attenuates mitochondrial Ca2+ uptake and also abolishes its positive correlation with ER-mitochondrion proximity. We postulate that during IP3 -induced Ca2+ release, Ca2+ uptake is confined to ER-close mitochondria, because of the simultaneous activation of the protein kinases. Attenuation of Ca2+ uptake prevents Ca2+ overload of mitochondria and thus protects the cell against apoptosis. On the other hand, all the mitochondria accumulate Ca2+ at a non-inhibited rate during physiological Ca2+ influx through the plasma membrane. Membrane potential is higher in ER-distant mitochondria, providing a bigger driving force for Ca2+ uptake. Our model explains why comparable mitochondrial Ca2+ signals are formed in response to K+ and angiotensin II (equipotent in respect to global cytosolic Ca2+ signals), although only the latter generates high-Ca2+ microdomains. [source] The formation of eclogite facies metatroctolites and a general petrogenetic grid in Na2O,CaO,FeO,MgO,Al2O3,SiO2,H2O (NCFMASH)JOURNAL OF METAMORPHIC GEOLOGY, Issue 9 2002G. Rebay Abstract Eclogite facies metatroctolites from a variety of Western Alps localities (Voltri, Monviso, Lanzo, Allalin, Zermat,Saas, etc.) that preserve textural evidence of their original form as bimineralic olivine-plagioclase rocks are considered in terms of calculated mineral equilibria in the system Na2O-CaO-FeO-MgO-Al2O3 -SiO2 -H2O (NCFMASH). Pseudosections, based on a new petrogenetic grid for NCFMASH presented here, are used to unravel the metamorphic history of the metatroctolites, considering the rocks to consist of different composition microdomains corresponding to the original olivine and plagioclase grains. On the basis that the preservation of the mineral assemblage in each microdomain will tend to be from where on a rock's P,T path the metamorphic fluid phase is used up via rehydration reactions, P,T pseudosections contoured for water content, and P,T path-MH2O (amount of water) pseudosections, are used to examine fluid behaviour in each microdomain. We show that the different microdomains are likely to preserve their mineral assemblages from different places on the P,T path. For the olivine microdomain, the diagnostic mineral assemblage is chloritoid + talc (+ garnet + omphacite). The preservation of this assemblage, in the light of the closed system P,T path-MH2O relationships, implies that the microdomain loses its metamorphic fluid as it starts to decompress, and, in the absence of subsequent hydration, the high pressure mineral assemblage is then preserved. In the plagioclase microdomain, the diagnostic assemblage is epidote (or zoisite) + kyanite + quartz suggesting a lower pressure (of about 2 GPa) than for the olivine microdomain. In the light of P,T path-MH2O relationships, development of this assemblage implies breakdown of lawsonite across the lawsonite breakdown reaction, regardless of the maximum pressure reached. It is likely that the plagioclase microdomain was mainly fluid-absent prior to lawsonite breakdown, only becoming fluid-present across the reaction, then immediately becoming fluid-absent again. [source] Role of group A Streptococcus HtrA in the maturation of SpeB proteasePROTEINS: STRUCTURE, FUNCTION AND BIOINFORMATICS, Issue 24 2007Jason N. Cole Abstract The serine protease high-temperature requirement A (HtrA) (DegP) of the human pathogen Streptococcus pyogenes (group A Streptococcus; GAS) is localized to the ExPortal secretory microdomain and is reportedly essential for the maturation of cysteine protease streptococcal pyrogenic exotoxin B (SpeB). Here, we utilize HSC5 (M5 serotype) and the in-frame isogenic mutant HSC5,htrA to determine whether HtrA contributes to the maturation of other GAS virulence determinants. Mutanolysin cell wall extracts and secreted proteins were arrayed by 2-DE and identified by MALDI-TOF PMF analysis. HSC5,htrA had elevated levels of cell wall-associated M protein, whilst the supernatant had higher concentrations of M protein fragments and a reduced amount of mature SpeB protease, compared to wild-type (WT). Western blot analysis and protease assays revealed a delay in the maturation of SpeB in the HSC5,htrA supernatant. HtrA was unable to directly process SpeB zymogen (proSpeB) to the active form in vitro. We therefore conclude that HtrA plays an indirect role in the maturation of cysteine protease SpeB. [source] Ca2+ microdomains near plasma membrane Ca2+ channels: impact on cell functionTHE JOURNAL OF PHYSIOLOGY, Issue 13 2008Anant B. Parekh In eukaryotic cells, a rise in cytoplasmic Ca2+ can activate a plethora of responses that operate on time scales ranging from milliseconds to days. Inherent to the use of a promiscuous signal like Ca2+ is the problem of specificity: how can Ca2+ activate some responses but not others? We now know that the spatial profile of the Ca2+ signal is important Ca2+ does not simply rise uniformly throughout the cytoplasm upon stimulation but can reach very high levels locally, creating spatial gradients. The most fundamental local Ca2+ signal is the Ca2+ microdomain that develops rapidly near open plasmalemmal Ca2+ channels like voltage-gated L-type (Cav1.2) and store-operated CRAC channels. Recent work has revealed that Ca2+ microdomains arising from these channels are remarkably versatile in triggering a range of responses that differ enormously in both temporal and spatial profile. Here, I delineate basic features of Ca2+ microdomains and then describe how these highly local signals are used by Ca2+ -permeable channels to drive cellular responses. [source] cAMP microdomains and L-type Ca2+ channel regulation in guinea-pig ventricular myocytesTHE JOURNAL OF PHYSIOLOGY, Issue 3 2007Sunita Warrier Many different receptors can stimulate cAMP synthesis in the heart, but not all elicit the same functional responses. For example, it has been recognized for some time that prostaglandins such as PGE1 increase cAMP production and activate PKA, but they do not elicit responses like those produced by ,-adrenergic receptor (,AR) agonists such as isoproterenol (isoprenaline), even though both stimulate the same signalling pathway. In the present study, we confirm that isoproterenol, but not PGE1, is able to produce cAMP-dependent stimulation of the L-type Ca2+ current in guinea pig ventricular myocytes. This is despite finding evidence that these cells express EP4 prostaglandin receptors, which are known to activate Gs -dependent signalling pathways. Using fluorescence resonance energy transfer-based biosensors that are either freely diffusible or bound to A kinase anchoring proteins, we demonstrate that the difference is due to the ability of isoproterenol to stimulate cAMP production in cytosolic and caveolar compartments of intact cardiac myocytes, while PGE1 only stimulates cAMP production in the cytosolic compartment. Unlike other receptor-mediated responses, compartmentation of PGE1 responses was not due to concurrent activation of a Gi -dependent signalling pathway or phosphodiesterase activity. Instead, compartmentation of the PGE1 response in cardiac myocytes appears to be due to transient stimulation of cAMP in a microdomain that can communicate directly with the bulk cytosolic compartment but not the caveolar compartment associated with ,AR regulation of L-type Ca2+ channel function. [source] Large Bore Catheters with Surface Treatments versus Untreated Catheters for Vascular Access in HemodialysisARTIFICIAL ORGANS, Issue 7 2004Rolf Bambauer Abstract:, Infection, thrombosis, and stenosis are among the most frequent complications associated with blood-contacting catheters. Complications resulting from infection remain a major problem for hemodialysis catheters, with significant numbers of catheters being removed due to catheter-related sepsis. Numerous strategies have been employed to reduce the occurrence of infection and im-prove long-term outcomes, with varying degrees of success. The most important is the careful and sterile handling by the attending staff of the catheters during hemodialysis treatments to minimize or stop a microbial colonization of the skin and the catheter. Another approach is coating the external surface of the catheters with substances which are antibacterial like silver and/or substances with low thrombogenicity like silicone. This investigation reviews results of animal and clinical experiments conducted to assess the efficacy and biocompatibility of silver and silicone coated dialysis catheters. It is concluded that silver coatings can reduce bacterial colonization and occurrence of infection associated with these devices. The catheters employing ion implantation of silicone rubber showed low thrombogenicity. Results of the studies indicate that ion beam based processes can be used to improve thrombus and infection resistance of blood contacting catheters. A new development is the microdomain structured surface (PUR-SMA coated catheters). Preliminary results with these catheters are very encouraging. [source] GM1,/,GD1b,/,GA1 synthase expression results in the reduced cancer phenotypes with modulation of composition and raft-localization of gangliosides in a melanoma cell lineCANCER SCIENCE, Issue 9 2010Yu Dong Gangliosides are expressed in neuroectoderm-derived tumors, and seemed to play roles in the regulation of cancer properties. To examine the behavior and roles of individual gangliosides, GM1/GD1b/GA1 synthase cDNA was introduced into the melanoma cell line SK-MEL-37, and changes in tumor phenotypes were analyzed. The transfectant cells showed neo-expression of GD1b, GT1b, and GM1, and reduced expression of GM3, GM2, GD2, and GD3. Function analyses revealed that the transfectant cells had definite reduction in cell growth and invasion. Tyrosine-phosphorylation levels of proteins such as p130Cas and paxillin were also reduced in the transfectants. These results suggested that the expression of GM1/GD1b/GA1 synthase resulted in the suppression of tumor properties. In the analyses of the floating patterns of gangliosides using fractions from sucrose density gradient ultracentrifugation of TritonX-100 extracts, the majority of gangliosides were found in glycolipid-enriched microdomain (GEM)/raft fractions, while GD3, GD1b, and GT1b in the transfectant cells tended to disperse to non-GEM/raft fractions. Furthermore, GD3, GD1b, and GT1b in non-GEM/raft dominantly had unsaturated fatty acids, while those in GEM/rafts contained more saturated forms than in non-GEM/rafts. This might be a mechanism for the decreased tumor properties in the transfectants of GM1/GD1b/GA1 synthase cDNA. (Cancer Sci 2010) [source] Modulation of calcium signalling by intracellular organelles seen with targeted aequorinsACTA PHYSIOLOGICA, Issue 1 2009M. T. Alonso Abstract The cytosolic Ca2+ signals that trigger cell responses occur either as localized domains of high Ca2+ concentration or as propagating Ca2+ waves. Cytoplasmic organelles, taking up or releasing Ca2+ to the cytosol, shape the cytosolic signals. On the other hand, Ca2+ concentration inside organelles is also important in physiology and pathophysiology. Comprehensive study of these matters requires to measure [Ca2+] inside organelles and at the relevant cytosolic domains. Aequorins, the best-known chemiluminescent Ca2+ probes, are excellent for this end as they do not require stressing illumination, have a large dynamic range and a sharp Ca2+ -dependence, can be targeted to the appropriate location and engineered to have the proper Ca2+ affinity. Using this methodology, we have evidenced the existence in chromaffin cells of functional units composed by three closely interrelated elements: (1) plasma membrane Ca2+ channels, (2) subplasmalemmal endoplasmic reticulum and (3) mitochondria. These Ca2+ -signalling triads optimize Ca2+ microdomains for secretion and prevent propagation of the Ca2+ wave towards the cell core. Oscillatory cytosolic Ca2+ signals originate also oscillations of mitochondrial Ca2+ in several cell types. The nuclear envelope slows down the propagation of the Ca2+ wave to the nucleus and filters high frequencies. On the other hand, inositol-trisphosphate may produce direct release of Ca2+ to the nucleoplasm in GH3 pituitary cells, thus providing mechanisms for selective nuclear signalling. Aequorins emitting at different wavelengths, prepared by fusion either with green or red fluorescent protein, permit simultaneous and independent monitorization of the Ca2+ signals in different subcellular domains within the same cell. [source] When is high-Ca2+ microdomain required for mitochondrial Ca2+ uptake?,ACTA PHYSIOLOGICA, Issue 1 2009A. Spät Abstract Ca2+ release from IP3 -sensitive stores in the endoplasmic reticulum (ER) induced by Ca2+ -mobilizing agonists generates high-Ca2+ microdomains between ER vesicles and neighbouring mitochondria. Here we present a model that describes when such microdomains are required and when submicromolar [Ca2+] is sufficient for mitochondrial Ca2+ uptake. Mitochondrial Ca2+ uptake rate in angiotensin II-stimulated H295R adrenocortical cells correlates with the proximity between ER vesicles and the mitochondrion, reflecting the uptake promoting effect of high-Ca2+ peri-mitochondrial microdomains. Silencing or inhibition of p38 mitogen-activated protein kinase (MAPK) or inhibition of the novel isoforms of protein kinase C enhances mitochondrial Ca2+ uptake and abolishes the positive correlation between Ca2+ uptake and ER-mitochondrion proximity. Inhibition of protein phosphatases attenuates mitochondrial Ca2+ uptake and also abolishes its positive correlation with ER-mitochondrion proximity. We postulate that during IP3 -induced Ca2+ release, Ca2+ uptake is confined to ER-close mitochondria, because of the simultaneous activation of the protein kinases. Attenuation of Ca2+ uptake prevents Ca2+ overload of mitochondria and thus protects the cell against apoptosis. On the other hand, all the mitochondria accumulate Ca2+ at a non-inhibited rate during physiological Ca2+ influx through the plasma membrane. Membrane potential is higher in ER-distant mitochondria, providing a bigger driving force for Ca2+ uptake. Our model explains why comparable mitochondrial Ca2+ signals are formed in response to K+ and angiotensin II (equipotent in respect to global cytosolic Ca2+ signals), although only the latter generates high-Ca2+ microdomains. [source] Plectin deposition at podosome rings requires myosin contractilityCYTOSKELETON, Issue 8 2008Annica Gad Abstract Metalloproteinase-dependent tissue invasion requires the formation of podosomes and invadopodia for localized matrix degradation. Actin cytoskeleton remodeling via Arp2/3-mediated actin polymerization is essential for podosome formation, and dynamic microtubules have an important role in maintaining podosome turnover in macrophages and osteoclasts. Little is known, however, about the involvement of the intermediate filament cytoskeleton in formation, stabilization, and turnover of podosomes. Here we show that vimentin intermediate filaments colocalize with the early sites of podosome formation at the stress fiber - focal adhesion interface in cultured vascular smooth muscle cells, but do not directly contribute to podosome formation, or stabilization. In unstimulated A7r5 cells the cytolinker protein plectin poorly colocalized with vimentin and the microdomains, but following induction by phorbol ester accumulated in the rings that surround the podosomes. In plectin-deficient A7r5 cells actin stress fiber remodelling is reduced in response to PDBu, and small podosomes remain localized at stable actin stress fibres. Pharmacological inhibition of actomyosin contractility by blebbistatin leads to an aberrant localization of podosomes away from the cell periphery and induces failure of plectin to surround the outer perimeter of these invasive adhesions. Taken together, we conclude that plectin is involved in growth and maturation of podosomes by reducing focal adhesion and stress fiber turnover, and that actomyosin-dependent contractility is required for the peripheral localization and specific deposition of plectin at the podosome rings. Cell Motil. Cytoskeleton 2008. © 2008 Wiley-Liss, Inc. [source] Abnormal CTLA-4 function in T cells from patients with systemic lupus erythematosusEUROPEAN JOURNAL OF IMMUNOLOGY, Issue 2 2010Elizabeth C. Jury Abstract CTLA-4 is a critical gatekeeper of T-cell activation and immunological tolerance and has been implicated in patients with a variety of autoimmune diseases through genetic association. Since T cells from patients with the autoimmune disease systemic lupus erythematosus (SLE) display a characteristic hyperactive phenotype, we investigated the function of CTLA-4 in SLE. Our results reveal increased CTLA-4 expression in FOXP3, responder T cells from patients with SLE compared with other autoimmune rheumatic diseases and healthy controls. However, CTLA-4 was unable to regulate T-cell proliferation, lipid microdomain formation and phosphorylation of TCR-, following CD3/CD28 co-stimulation, in contrast to healthy T cells. Although lupus T cells responded in vitro to CD3/CD28 co-stimulation, there was no parallel increase in CTLA-4 expression, which would normally provide a break on T-cell proliferation. These defects were associated with exclusion of CTLA-4 from lipid microdomains providing an anatomical basis for its loss of function. Collectively our data identify CTLA-4 dysfunction as a potential cause for abnormal T-cell activation in patients with SLE, which could be targeted for therapy. [source] Recombinase-deficient T cell development by selective accumulation of CD3 into lipid raftsEUROPEAN JOURNAL OF IMMUNOLOGY, Issue 4 2008Denise Ferrera Abstract The pre-T cell receptor (pre-TCR) promotes the development of thymocytes with productive rearrangement at the TCR ,,chain locus by signaling in a ligand-independent fashion. The TCR ,,chain associates with the invariant pre-T, (pT,) chain, which bears specific charged residues in the extracellular portion mediating pre-TCR self-oligomerization. In recombinase-deficient thymocytes, calnexin (CNX) associated with CD3 chains is inefficiently retained in the endoplasmic reticulum (ER) and weakly expressed in the plasma membrane. Deliberate cross-linking of CNX/CD3 complexes mimics pre-TCR signaling. Here, we show that, analogously to the pT, chain, surface CNX is palmitoylated and that CD3 prominently accumulated in lipid rafts upon cross-linking. Mutant CNX isoforms devoid of ER retention determined pre-TCR-like signaling and simulated ,,selection only when stably translocating CD3 to lipid rafts. Inclusion of the palmitoylated cytoplasmic tail from the pT, chain in recombinant CNX strikingly improved the pre-TCR-like signaling efficiency of CNX/CD3 in rafts. This study indicates that lipid rafts in the plasma membrane represent proficient microdomains for the initiation of pre-TCR signaling, and supports the view that ,,selection by oligomerized pre-TCR is implemented by the pT, cytoplasmic tail. [source] Characterization of Xenopus egg membrane microdomains containing uroplakin Ib/III complex: roles of their molecular interactions for subcellular localization and signal transductionGENES TO CELLS, Issue 2 2007A.K.M. Mahbub Hasan A single-transmembrane protein uroplakin III (UPIII) and its tetraspanin binding-partner uroplakin Ib (UPIb) are members of the UP proteins that were originally identified in mammalian urothelium. In Xenopus laevis eggs, these proteins: xUPIII and xUPIb, are components of the cholesterol-enriched membrane microdomains or "rafts" and involved in the sperm,egg membrane interaction and subsequent egg activation signaling via Src tyrosine kinase at fertilization. Here, we investigate whether the xUPIII-xUPIb complex is in close proximity to CD9, a tetraspanin that has been implicated in the sperm,egg fusion in the mouse and GM1, a ganglioside typically enriched in egg rafts. Preparation of the egg membrane microdomains using different non-ionic detergents (Brij 98 and Triton X-100), chemical cross-linking, co-immunoprecipitation, in vitro kinase assay and in vitro fertilization experiments demonstrated that GM1, but not CD9, is in association with the xUPIII-xUPIb complex and contributes to the sperm-dependent egg activation. Transfection experiments using HEK293 cells demonstrated that xUPIII and xUPIb localized efficiently to the cholesterol-dependent membrane microdomains when they were co-expressed, whereas co-expression of xUPIII and CD9, instead of xUPIb, did not show this effect. Furthermore, xUPIII and xUPIb were shown to suppress kinase activity of the wild type, but not a constitutively active form of, Xenopus Src protein co-expressed in HEK293 cells. These results provide novel insight into the molecular architecture of the egg membrane microdomains containing xUPIII, xUPIb and Src, which may contribute to the understanding of sperm,egg interaction and signaling during Xenopus fertilization. [source] Phosphorylation and lipid raft association of fibroblast growth factor receptor-2 in oligodendrocytes,GLIA, Issue 9 2009M. R. Bryant Abstract Fibroblast growth factors (FGFs) and their receptors (FGFRs) initiate diverse cellular responses that contribute to the regulation of oligodendrocyte (OL) function. To understand the mechanisms by which FGFRs elicit these cellular responses, we investigated the phosphorylation of signal transduction proteins and the role of cholesterol-glycosphingolipid-enriched "lipid raft" microdomains in differentiated OLs. Surprisingly, we found that the most abundant tyrosine-phosphorylated protein in OLs was the 120-kd isoform of FGFR2 and that it was phosphorylated even in the absence of FGF2, suggesting a potential ligand-independent function for this receptor. Furthermore, FGFR2, but not FGFR1, was associated with lipid raft microdomains in OLs and myelin (but not in astrocytes). This provides the first evidence for the association of FGFR with TX-100-insoluble lipid raft fractions. FGFR2 phosphorylated the key downstream target, FRS2 in OLs. Raft disruption resulted in loss of phosphorylated FRS2 from lipid rafts, coupled with the loss of Akt but not of Mek or Erk phosphorylation. This suggests that FGFR2-FRS2 signaling in lipid rafts operates via the PI3-Kinase/Akt pathway rather than the Ras/Mek/Erk pathway, emphasizing the importance of microenvironments within the cell membrane. Also present in lipid rafts in OLs and myelin, but not in astrocytes, was a novel 52-kd isoform of FGFR2 that lacked the extracellular ligand-binding region. These results demonstrate that FGFR2 in OLs and myelin possess unique characteristics that are specific both to receptor type and to OLs and provide a novel mechanism to elicit distinct cellular responses that mediate both FGF-dependent and -independent functions. © 2008 Wiley-Liss, Inc. [source] Kir4.1 and AQP4 associate with Dp71- and utrophin-DAPs complexes in specific and defined microdomains of Müller retinal glial cell membraneGLIA, Issue 6 2008Patrice E. Fort Abstract The dystrophin-associated proteins (DAPs) complex consisting of dystroglycan, syntrophin, dystrobrevin, and sarcoglycans in muscle cells is associated either with dystrophin or its homolog utrophin. In rat retina, a similar complex was found associated with dystrophin-Dp71 that serves as an anchor for the inwardly rectifying potassium channel Kir4.1 and the aqueous pore, aquaporin-4 (AQP4). Here, using immunofluorescence imaging of isolated retinal Müller glial cells and co-immunoprecipitation experiments performed on an enriched Müller glial cells end-feet fraction, we investigated the effect of Dp71 deletion on the composition, anchoring, and membrane localization of the DAPs,Kir4.1 and/or ,AQP4 complex. Two distinct complexes were identified in the end-feet fraction associated either with Dp71 or with utrophin. Upon Dp71 deletion, the corresponding DAPs complex was disrupted and a compensating utrophin upregulation was observed, accompanied by diffuse overall staining of Kir4.1 along the Müller glial cells and redistribution of the K+ conductance. Dp71 deficiency was also associated with a marked reduction of AQP4 and ,-dystroglycan expression. Furthermore, it was observed that the Dp71,DAPs dependent complex could be, at least partially, associated with a specific membrane fraction. These results demonstrate that Dp71 has a central role in the molecular scaffold responsible for anchoring AQP4 and Kir4.1 in Müller cell end-feet membranes. They also show that despite its close relationship to the dystrophin proteins and its correlated upregulation, utrophin is only partially compensating for the absence of Dp71 in Müller glial cells. © 2008 Wiley-Liss, Inc. [source] Rafts in oligodendrocytes: Evidence and structure,function relationshipGLIA, Issue 6 2006Ellen Gielen Abstract The plasma membrane of eukaryotic cells exhibits lateral inhomogeneities, mainly containing cholesterol and sphingomyelin, which provide liquid-ordered microdomains (lipid "rafts") that segregate membrane components. Rafts are thought to modulate the biological functions of molecules that become associated with them, and as such, they appear to be involved in a variety of processes, including signal transduction, membrane sorting, cell adhesion and pathogen entry. Although still a matter of ongoing debate, evidence in favor of the presence of these microdomains is gradually accumulating but a consensus on issues like their size, lifetime, composition, and biological significance has yet to be reached. Here, we provide an overview of the evidence supporting the presence of rafts in oligodendrocytes, the myelin-producing cells of the central nervous system, and discuss their functional significance. The myelin membrane differs fundamentally from the plasma membrane, both in lipid and protein composition. Moreover, since myelin membranes are unusually enriched in glycosphingolipids, questions concerning the biogenesis and functional relevance of microdomains thus appear of special interest in oligodendrocytes. The current picture of rafts in oligodendrocytes is mainly based on detergent methods. The robustness of such data is discussed and alternative methods that may provide complementary data are indicated. © 2006 Wiley-Liss, Inc. [source] Directed Helical Growth: A Spring-Like Behavior of Chiral Block Copolymer with Helical Nanostructure Driven by Crystallization (Adv. Funct.ADVANCED FUNCTIONAL MATERIALS, Issue 3 2009Mater. Crystalline helices (PLLA crystallization directed by helical confined microdomains) and crystalline cylinders (phase transformation of helical nano structures dictated by crystallization) are obtained by controlling the crystallization temperature of PLLA with respect to the glass transition temperature of PS in PS-PLLA block copolymers; this process is described by J.-W. Chiang et al. on page 448. A spring-like behavior of the PLLA helical nanostructures embedded in the PS matrix can be driven by crystallization, so as to dictate the transformation of the helices, resulting in crystalline cylinders that might represent a possible avenue for the design of switchable large-strain actuators. [source] A Spring-Like Behavior of Chiral Block Copolymer with Helical Nanostructure Driven by CrystallizationADVANCED FUNCTIONAL MATERIALS, Issue 3 2009Yeo-Wan Chiang Abstract The crystallization of helical nanostructure resulting from the self-assembly of a chiral diblock copolymer, poly(styrene)- b -poly(L -lactide) (PS-PLLA), is studied. Various crystalline PS-PLLA nanostructures are obtained by controlling the crystallization temperature of PLLA (Tc,PLLA), at which crystalline helices and crystalline cylinders occur while Tc,PLLA,<,Tg,PS (the glass transition temperature of PS) and Tc,PLLA,,,Tg,PS, respectively. As evidenced by selected-area electron diffraction and two-dimensional X-ray diffraction results, the PLLA crystallites under confinement reveal a unique anisotropic character regardless of the crystallization temperature. On the basis of observed uniaxial scattering results the PLLA crystallites grown within the microdomains are identified as crystals with preferential growth directions either along the [100] or along the [110]-axes of the PLLA crystalline unit cell, at which the molecular chains and the growth direction are normal and parallel to the central axes of helices, respectively. The formation of this exclusive crystalline growth is attributed to the spatial confinement effect for crystallization. While Tc,PLLA,<,Tg,PS, owing to the directed crystallization by helical confinement, the preferential crystalline growth leads to the crystallization following a helical track with growth direction parallel to the central axes of helices through a twisting mechanism. Consequently, winding crystals with specific crystallographic orientation within the helical microdomains can be found. By contrast, while Tc,PLLA,,,Tg,PS, the preferential growth may modulate the curvature of microdomains by shifting the molecular chains to access the fast path for crystalline growth due to the increase in chain mobility. As a result, a spring-like behavior of the helical nanostructure can be driven by crystallization so as to dictate the transformation of helices, resulting in crystalline cylinders that might be applicable to the design of switchable large-strain actuators. [source] Directed Self-Assembly of Block Copolymers on Two-Dimensional Chemical Patterns Fabricated by Electro-Oxidation NanolithographyADVANCED MATERIALS, Issue 20 2010Ji Xu A hexagonal web of carboxylic-terminated nanostripes (left image, bright areas) is patterned onto a methyl-terminated surface of an octadecyltrichlorosilane monolayer. A thermally annealed polystyrene- block -poly(ethylene oxide) (PS- b -PEO) thin-film, spin-cast on the chemical pattern (right image), exhibits surface normal oriented cylindrical PEO microdomains on the methyl-terminated regions only. These chemical patterns effectively template the order and spatial orientation of diblock-copolymer microdomains. [source] Initiation of TCR signaling: regulation within CD3 dimersIMMUNOLOGICAL REVIEWS, Issue 1 2003Balbino Alarcón Summary: The number of possible T cell activation outcomes resulting from T cell receptor (TCR) engagement suggests that the TCR is able to differentially activate a myriad of signaling pathways depending on the nature of the stimulus. The complex structural organization of the TCR itself could underlie this diversity of responses. Assembly and stoichiometric studies have helped us to shed some light on the initiation of TCR signaling. The TCR is composed of TCR and CD3 dimers. Changes in the interaction between CD3 subunits within the CD3 dimers and in the interaction of these dimers with the TCR heterodimer could be the triggering mechanism that initiates the first activation events. One of the hallmarks of these early changes in TCR conformation is the induced recruitment of the adapter protein Nck to a proline-rich sequence of the cytoplasmic tail of CD3,, but there may be others. According to our most recent observations, the TCR is organized in pre-existing clusters within plasma membrane microdomains, exhibiting a complexity above and beyond that of dimer composition complexity. How the presence of TCR in clusters influences TCR avidity and propagation of TCR signals is something that has yet to be investigated. [source] Regulation of T-cell receptor signalling by membrane microdomainsIMMUNOLOGY, Issue 4 2004Tahir M. Razzaq Summary There is now considerable evidence suggesting that the plasma membrane of mammalian cells is compartmentalized by functional lipid raft microdomains. These structures are assemblies of specialized lipids and proteins and have been implicated in diverse biological functions. Analysis of their protein content using proteomics and other methods revealed enrichment of signalling proteins, suggesting a role for these domains in intracellular signalling. In T lymphocytes, structure/function experiments and complementary pharmacological studies have shown that raft microdomains control the localization and function of proteins which are components of signalling pathways regulated by the T-cell antigen receptor (TCR). Based on these studies, a model for TCR phosphorylation in lipid rafts is presented. However, despite substantial progress in the field, critical questions remain. For example, it is unclear if membrane rafts represent a homogeneous population and if their structure is modified upon TCR stimulation. In the future, proteomics and the parallel development of complementary analytical methods will undoubtedly contribute in further delineating the role of lipid rafts in signal transduction mechanisms. [source] CD20-mediated apoptosis: signalling through lipid raftsIMMUNOLOGY, Issue 2 2002Julie P. Deans Summary CD20 is an effective target for therapeutic B-cell depletion with monoclonal antibodies. One proposed mechanism of action is direct cytotoxicity mediated via tyrosine kinase-dependent signalling pathways activated upon CD20 cross-linking. The association of CD20 with membrane microdomains known as lipid rafts, enriched in src-family tyrosine kinases and other signalling effectors, suggests an indirect mechanism of anti-CD20-induced apoptosis in which activation of src-family kinases occurs as a consequence of lipid raft clustering. [source] Fabrication of Highly Ordered Silicon Oxide Dots and Stripes from Block Copolymer Thin Films,ADVANCED MATERIALS, Issue 4 2008S. Park A general route to fabricate highly ordered arrays of nanoscopic silicon oxide dots and stripes (see figure) from block copolymer thin films is described. Poly(styrene- b -4-vinylpyridine) thin films with cylindrical microdomains oriented normal and parallel to the surface were used as templates for the fabrication of nanoscopic silicon oxide, with polydimethylsiloxane as the inorganic precursor. [source] Self-Assembly of Nanoparticle,Copolymer Mixtures: A Kinetic Point of View,ADVANCED MATERIALS, Issue 3 2007J. He The prediction of synergistic effects between two self-organizing systems is tested. In,situ grazing-incidence small-angle X-ray scattering (see figure) is used during thermal annealing of a nanoparticle,copolymer mixture, and shows that the orientation of the microdomains begins at the free surface and propagates in the film towards the substrate. This synergistic interaction is shown to apply to both cylindrical and lamellar block-copolymer morphologies. [source] Membrane Photolithography: Direct Micropatterning and Manipulation of Fluid Phospholipid Membranes in the Aqueous Phase Using Deep-UV Light,ADVANCED MATERIALS, Issue 14 2004K. Yee A wet photolithography approach using light-activated, localized, oxidative chemistry can directly pattern fluid phospholipid bilayers submerged in aqueous phases. Targeted incorporation of secondary components within pattern voids (see Figure) allows many membrane dynamical processes to be probed and optically defined arrays of holes, functional membrane microdomains, and proteins embedded in a lipidic background can be designed. [source] Low-density caveolae-like membrane from Xenopus laevis oocytes is enriched in RasJOURNAL OF CELLULAR BIOCHEMISTRY, Issue 1 2001Susan E. SadlerArticle first published online: 10 JUL 200 Abstract Detergent-free discontinuous sucrose density gradient centrifugation was used to resolve low- and high-density membrane fractions from Xenopus laevis oocytes. Compared to high-density membrane, low-density oocyte membrane is enriched two-fold in cholesterol and highly enriched in ganglioside GM1. Protein immunoblotting of membrane fractions from whole cells with polyclonal anti-human caveolin antibody detected multiple bands, including a distinctive triad with apparent molecular weights of 21, 33, and 48 kDa. To more clearly determine which of these caveolin-like protein(s) is associated with the oocyte plasma membrane, microdissection was used to separate external membrane (cortical preparations containing plasma membrane) from intracellular membrane. Cortical membrane preparations displayed a single 21-kDa caveolin-like protein in low-density membrane. Internal oocyte membrane displayed the higher molecular weight bands of 33 and 48 kDa and a lesser amount of the 21-kDa protein in low-density membrane fractions. Monoclonal anti-human Ras antibody detected a single 23-kDa immunoblot band that is enriched an average of eight-fold in low-density membrane fractions prepared from whole cells. This is the first report of caveolin-associated, low-density membrane in amphibian oocytes, and is consistent with a role for caveolin and caveolae-like microdomains in oocyte signal transduction. © 2001 Wiley-Liss, Inc. [source] Multiple Kv1.5 targeting to membrane surface microdomains,JOURNAL OF CELLULAR PHYSIOLOGY, Issue 3 2008Ramón Martínez-Mármol Surface expression of voltage-dependent K+ channels (Kv) has a pivotal role in leukocyte physiology. Although little is known about the physiological role of lipid rafts, these microdomains concentrate signaling molecules and their ion channel substrates. Kv1.3 associates with Kv1.5 to form functional channels in macrophages. Different isoform stoichiometries lead to distinct heteromeric channels which may be further modulated by targeting the complex to different membrane surface microdomains. Kv1.3 targets to lipid rafts, whereas Kv1.5 localization is under debate. With this in mind, we wanted to study whether heterotetrameric Kv1.5-containing channels target to lipid rafts. While in transfected HEK-293 cells, homo- and heterotetrameric channels targeted to rafts, Kv1.5 did not target to rafts in macrophages. Therefore, Kv1.3/Kv1.5 hybrid channels are mostly concentrated in non-raft microdomains. However, LPS-induced activation, which increases the Kv1.3/Kv1.5 ratio and caveolin, targeted Kv1.5 back to lipid rafts. Moreover, Kv1.5 did not localize to low-buoyancy fractions in L6E9 skeletal myoblasts, which also coexpress both channels, heart membranes or cardiomyocyes. Coexpression of a Cav3DGV -mutant confined Kv1.5 to Cav3DGV -vesicles of HEK cells. Contrarily, coexpression of Kv,2.1 impaired the Kv1.5 targeting to raft microdomains in HEK cells. Our results indicate that Kv1.5 partnership interactions are underlying mechanisms governing channel targeting to lipid rafts. J. Cell. Physiol. 217: 667,673, 2008. © 2008 Wiley-Liss, Inc. [source] The formation of eclogite facies metatroctolites and a general petrogenetic grid in Na2O,CaO,FeO,MgO,Al2O3,SiO2,H2O (NCFMASH)JOURNAL OF METAMORPHIC GEOLOGY, Issue 9 2002G. Rebay Abstract Eclogite facies metatroctolites from a variety of Western Alps localities (Voltri, Monviso, Lanzo, Allalin, Zermat,Saas, etc.) that preserve textural evidence of their original form as bimineralic olivine-plagioclase rocks are considered in terms of calculated mineral equilibria in the system Na2O-CaO-FeO-MgO-Al2O3 -SiO2 -H2O (NCFMASH). Pseudosections, based on a new petrogenetic grid for NCFMASH presented here, are used to unravel the metamorphic history of the metatroctolites, considering the rocks to consist of different composition microdomains corresponding to the original olivine and plagioclase grains. On the basis that the preservation of the mineral assemblage in each microdomain will tend to be from where on a rock's P,T path the metamorphic fluid phase is used up via rehydration reactions, P,T pseudosections contoured for water content, and P,T path-MH2O (amount of water) pseudosections, are used to examine fluid behaviour in each microdomain. We show that the different microdomains are likely to preserve their mineral assemblages from different places on the P,T path. For the olivine microdomain, the diagnostic mineral assemblage is chloritoid + talc (+ garnet + omphacite). The preservation of this assemblage, in the light of the closed system P,T path-MH2O relationships, implies that the microdomain loses its metamorphic fluid as it starts to decompress, and, in the absence of subsequent hydration, the high pressure mineral assemblage is then preserved. In the plagioclase microdomain, the diagnostic assemblage is epidote (or zoisite) + kyanite + quartz suggesting a lower pressure (of about 2 GPa) than for the olivine microdomain. In the light of P,T path-MH2O relationships, development of this assemblage implies breakdown of lawsonite across the lawsonite breakdown reaction, regardless of the maximum pressure reached. It is likely that the plagioclase microdomain was mainly fluid-absent prior to lawsonite breakdown, only becoming fluid-present across the reaction, then immediately becoming fluid-absent again. [source] Melt-producing and melt-consuming reactions in the Achankovil cordierite gneisses, South IndiaJOURNAL OF METAMORPHIC GEOLOGY, Issue 6 2002B. Cenki Abstract Migmatitic cordierite gneisses within the Achankovil Zone (AZ) of southern Pan-African India record melt-producing and subsequent melt-consuming mineral reactions. Early mineral assemblages Bt-Sil-Qtz and Bt-Sil-Spl, deduced from inclusion textures in garnet prophyroblasts, break down via successive dehydration melting reactions to high- T phase assemblages (e.g. Grt-Crd-Liq, Opx-Liq, Spl-Crd-Liq). Later back reactions between the restite and the in situ crystallizing melt resulted in thin cordierite coronas separating garnet from the leucosome, and partial resorption of garnet to Opx-Crd or Crd-Bt-Qtz symplectites. Leucosomes generally display a moderate (low-strain gneisses) to strong (high-strain gneisses) depletion of alkali feldspar attributed to mineral-melt back reactions partly controlled by the degree of melt segregation. Using a KFMASH partial petrogenetic grid that includes a melt phase, and qualitative pseudosections for microdomains of high and low Al/Si ratios, the successive phase assemblages and reaction textures are interpreted in terms of a clockwise P,T path culminating at about 6,7 kbar and 900,950 °C. This P,T path is consistent with, but more detailed than published results, which suggests that taking a melt phase into account is not only a valid, but also a useful approach. Comparing P,T data and lithological and isotopic data for the AZ with adjacent East Gondwana fragments, suggests the presence of a coherent metasedimentary unit exposed from southern Madagascar via South India (AZ) and Sri Lanka (Wanni Complex) to the Lützow,Holm Bay in Eastern Antarctica. [source] Microstructural tectonometamorphic processes and the development of gneissic layering: a mechanism for metamorphic segregationJOURNAL OF METAMORPHIC GEOLOGY, Issue 1 2000Williams The Mary granite, in the East Athabasca mylonite triangle, northern Saskatchewan, provides an example and a model for the development of non-migmatitic gneissic texture. Gneissic compositional layering developed through the simultaneous evolution of three microdomains corresponding to original plagioclase, orthopyroxene and matrix in the igneous rocks. Plagioclase phenocrysts were progressively deformed and recrystallized, first into core and mantle structures, and ultimately into plagioclase-rich layers or ribbons. Garnet preferentially developed in the outer portions of recrystallized mantles, and, with further deformation, produced garnet-rich sub-layers within the plagioclase-rich gneissic domains. Orthopyroxene was replaced by clinopyroxene and garnet (and hornblende if sufficient water was present), which were, in turn, drawn into layers with new garnet growth along the boundaries. The igneous matrix evolved through a number of transient fabric stages involving S-C fabrics, S-C-C, fabrics, and ultramylonitic domains. In addition, quartz veins were emplaced and subsequently deformed into quartz-rich gneissic layers. Moderate to highly strained samples display extreme mineralogical (compositional) segregation, yet most domains can be directly related to the original igneous precursors. The Mary granite was emplaced at approximately 900 °C and 1.0 GPa and was metamorphosed at approximately 750 °C and 1.0 GPa. The igneous rocks crystallized in the medium-pressure granulite field (Opx,Pl) but were metamorphosed on cooling into the high-pressure (Grt,Cpx,Pl) granulite field. The compositional segregation resulted from a dynamic, mutually reinforcing interaction between deformation, metamorphic and igneous processes in the deep crust. The production of gneissic texture by processes such as these may be the inevitable result of isobaric cooling of igneous rocks within a tectonically active deep crust. [source] | |||||||||||||||||||||||||||||||