scholarly journals Cell Surface Orifices of Caveolae and Localization of Caveolin to the Necks of Caveolae in Adipocytes

2003 ◽  
Vol 14 (10) ◽  
pp. 3967-3976 ◽  
Author(s):  
Hans Thorn ◽  
Karin G. Stenkula ◽  
Margareta Karlsson ◽  
Unn Örtegren ◽  
Fredrik H. Nystrom ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Cell Surface ◽  
Human Fibroblasts ◽  
High Resolution Electron Microscopy ◽  
Proximal Region ◽  
Cholesterol Depletion ◽  
Caveolin 1 ◽  
Resolution Electron ◽  
20 Nm ◽  
Membrane Proximal Region

Caveolae are noncoated invaginations of the plasma membrane that form in the presence of the protein caveolin. Caveolae are found in most cells, but are especially abundant in adipocytes. By high-resolution electron microscopy of plasma membrane sheets the detailed structure of individual caveolae of primary rat adipocytes was examined. Caveolin-1 and -2 binding was restricted to the membrane proximal region, such as the ducts or necks attaching the caveolar bulb to the membrane. This was confirmed by transfection with myc-tagged caveolin-1 and -2. Essentially the same results were obtained with human fibroblasts. Hence caveolin does not form the caveolar bulb in these cells, but rather the neck and may thus act to retain the caveolar constituents, indicating how caveolin participates in the formation of caveolae. Caveolae, randomly distributed over the plasma membrane, were very heterogeneous, varying in size between 25 and 150 nm. There was about one million caveolae in an adipocyte, which increased the surface area of the plasma membrane by 50%. Half of the caveolae, those larger than 50 nm, had access to the outside of the cell via ducts and 20-nm orifices at the cell surface. The rest of the caveolae, those smaller than 50 nm, were not open to the cell exterior. Cholesterol depletion destroyed both caveolae and the cell surface orifices.

scholarly journals Caveolin-1 is incorporated into mature respiratory syncytial virus particles during virus assembly on the surface of virus-infected cells

2002 ◽  
Vol 83 (3) ◽  
pp. 611-621 ◽  
Author(s):  
Gaie Brown ◽  
James Aitken ◽  
Helen W. McL. Rixon ◽  
Richard J. Sugrue
Keyword(s):  
Electron Microscopy ◽  
Plasma Membrane ◽  
Respiratory Syncytial Virus ◽  
Confocal Microscopy ◽  
Cell Surface ◽  
Filamentous Form ◽  
Caveolin 1 ◽  
Immuno Electron Microscopy ◽  
Infected Cells ◽  
Syncytial Virus

We have employed immunofluorescence microscopy and transmission electron microscopy to examine the assembly and maturation of respiratory syncytial virus (RSV) in the Vero cell line C1008. RSV matures at the apical cell surface in a filamentous form that extends from the plasma membrane. We observed that inclusion bodies containing viral ribonucleoprotein (RNP) cores predominantly appeared immediately below the plasma membrane, from where RSV filaments form during maturation at the cell surface. A comparison of mock-infected and RSV-infected cells by confocal microscopy revealed a significant change in the pattern of caveolin-1 (cav-1) fluorescence staining. Analysis by immuno-electron microscopy showed that RSV filaments formed in close proximity to cav-1 clusters at the cell surface membrane. In addition, immuno-electron microscopy showed that cav-1 was closely associated with early budding RSV. Further analysis by confocal microscopy showed that cav-1 was subsequently incorporated into the envelope of RSV filaments maturing on the host cell membrane, but was not associated with other virus structures such as the viral RNPs. Although cav-1 was incorporated into the mature virus, it was localized in clusters rather than being uniformly distributed along the length of the viral filaments. Furthermore, when RSV particles in the tissue culture medium from infected cells were examined by immuno-negative staining, the presence of cav-1 on the viral envelope was clearly demonstrated. Collectively, these findings show that cav-1 is incorporated into the envelope of mature RSV particles during egress.

Synthesis of ZnO2 Nanocrystals Produced by Hydrothermal Process

2009 ◽  
Vol 1242 ◽  
Author(s):  
R. Esparza ◽  
A. Aguilar ◽  
A. Escobedo-Morales ◽  
C. Patiño-Carachure ◽  
U. Pal ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Hydrogen Peroxide ◽  
Transmission Electron Microscopy ◽  
Hydrothermal Process ◽  
High Resolution Electron Microscopy ◽  
X Ray ◽  
Transmission Electron ◽  
Resolution Electron ◽  
Zinc Peroxide ◽  
20 Nm

ABSTRACTZinc peroxide (ZnO2) nanocrystals were directly produced by hydrothermal process. The nanocrystals were synthesized using zinc acetate as precursor and hydrogen peroxide as oxidant agent. The ZnO2 powders were characterized by X-ray powder diffraction and transmission electron microscopy. The results of transmission electron microscopy indicated that the ZnO2powders consisted of nanocrystals with diameters below to 20 nm and a faceted morphology. High resolution electron microscopy observations have been used in order to the structural characterization. ZnO2 nanocrystals exhibit a well-crystallized structure.

scholarly journals Regulation of caveolae through cholesterol-depletion dependent tubulation by PACSIN2/Syndapin II

2020 ◽  
Author(s):  
Aini Gusmira Amir ◽  
Kazuhiro Takemura ◽  
Kyoko Hanawa-Suetsugu ◽  
Kayoko Oono-Yakura ◽  
Kazuma Yasuhara ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Lipid Composition ◽  
Electrostatic Charge ◽  
Cholesterol Depletion ◽  
Membrane Deformation ◽  
Bar Domain ◽  
Caveolin 1 ◽  
Membrane Tubulation ◽  
Shaping Ability

AbstractThe membrane shaping ability of PACSIN2 via its FCH-BAR (F-BAR) domain has been shown to be essential for caveolar morphogenesis, presumably through the shaping of the caveolar neck. Caveolar membrane contains abundant levels of cholesterol. However, the role of cholesterol in PACSIN2-mediated membrane deformation remains unclear. We show that the binding of PACSIN2 to the membrane could be negatively regulated by the amount of cholesterol in the membrane. We prepared a reconstituted membrane based on the lipid composition of caveolae. The reconstituted membrane with cholesterol had a weaker affinity to the F-BAR domain of PACSIN2 than the membrane without cholesterol, presumably due to a decrease in electrostatic charge density. Consistently, the acute depletion of cholesterol from the plasma membrane resulted in the appearance of PACSIN2-localized tubules with caveolin-1 at their tips, suggesting that the presence of cholesterol inhibited the prominent membrane tubulation by PACSIN2. The tubules induced by PACSIN2 were suggested to be an intermediate of caveolae endocytosis. Consistently, the removal of caveolae from the plasma membrane upon cholesterol depletion was diminished in the cells deficient in PACSIN2. These data suggested that PACSIN2 mediated the caveolae internalization dependently on the amount of cholesterol at the plasma membrane, providing a possible mechanism for the cholesterol-dependent regulation of caveolae.

scholarly journals Clustering and Internalization of Toxic Amylin Oligomers in Pancreatic Cells Require Plasma Membrane Cholesterol

2011 ◽  
Vol 286 (41) ◽  
pp. 36086-36097 ◽  
Author(s):  
Saurabh Trikha ◽  
Aleksandar M. Jeremic
Keyword(s):  
Plasma Membrane ◽  
Cell Surface ◽  
Self Assembly ◽  
Surface Coverage ◽  
Islet Cells ◽  
Fold Increase ◽  
Water Soluble ◽  
Cholesterol Depletion ◽  
Β Cells ◽  
Pancreatic Hormone

Self-assembly of the human pancreatic hormone amylin into toxic oligomers and aggregates is linked to dysfunction of islet β-cells and pathogenesis of type 2 diabetes mellitus. Recent evidence suggests that cholesterol, an essential component of eukaryotic cells membranes, controls amylin aggregation on model membranes. However, the pathophysiological consequence of cholesterol-regulated amylin polymerization on membranes and biochemical mechanisms that protect β-cells from amylin toxicity are poorly understood. Here, we report that plasma membrane (PM) cholesterol plays a key role in molecular recognition, sorting, and internalization of toxic amylin oligomers but not monomers in pancreatic rat insulinoma and human islet cells. Depletion of PM cholesterol or the disruption of the cytoskeleton network inhibits internalization of amylin oligomers, which in turn enhances extracellular oligomer accumulation and potentiates amylin toxicity. Confocal microscopy reveals an increased nucleation of amylin oligomers across the plasma membrane in cholesterol-depleted cells, with a 2-fold increase in cell surface coverage and a 3-fold increase in their number on the PM. Biochemical studies confirm accumulation of amylin oligomers in the medium after depletion of PM cholesterol. Replenishment of PM cholesterol from intracellular cholesterol stores or by the addition of water-soluble cholesterol restores amylin oligomer clustering at the PM and internalization, which consequently diminishes cell surface coverage and toxicity of amylin oligomers. In contrast to oligomers, amylin monomers followed clathrin-dependent endocytosis, which is not sensitive to cholesterol depletion. Our studies identify an actin-mediated and cholesterol-dependent mechanism for selective uptake and clearance of amylin oligomers, impairment of which greatly potentiates amylin toxicity.

New nonhydrolytic route to synthesize crystalline BaTiO3 nanocrystals with surface capping ligands

2006 ◽  
Vol 21 (12) ◽  
pp. 3187-3195 ◽  
Author(s):  
Zhuoying Chen ◽  
Limin Huang ◽  
Jiaqing He ◽  
Yimei Zhu ◽  
Stephen O'Brien
Keyword(s):  
Electron Microscopy ◽  
Barium Titanate ◽  
Oleyl Alcohol ◽  
High Resolution Electron Microscopy ◽  
Decanoic Acid ◽  
Nonpolar Solvents ◽  
Selected Area Electron Diffraction ◽  
Resolution Electron ◽  
Capping Ligands ◽  
20 Nm

A new nonhydrolytic route for the preparation of well-crystallized size-tunable barium titanate (BaTiO3) nanocrystals capped with surface ligands is reported. Our approach involves: (i) synthesizing a “pseudo” bimetallic precursor, and (ii) combining the as-synthesized bimetallic precursor with a mixture of oleylamine with different surface coordinating ligands at 320 °C for crystallization and crystal growth. Different alcohols in the precursor synthesis and different carboxylic acids were used to study the effect of size and morphological control over the nanocrystals. Nanocrystals of barium titanate with diameters of 6–10 nm (capped with decanoic acid), 3–5 nm (capped with oleic acid), 10–20 nm (a nanoparticle and nanorod mixture capped with oleyl alcohol), and 2–3 nm (capped with oleyl alcohol) were synthesized, and can be easily dispersed into nonpolar solvents such as hexane or toluene. Techniques including x-ray diffraction, transmission electron microscopy, selected area electron diffraction, and high-resolution electron microscopy confirm the crystallinity and morphology of these as-synthesized nanocrystals.

Enlarged Crystalline Patches on the Plasma Membrane of Yeast Protoplasts

1980 ◽  
Vol 38 ◽  
pp. 686-687
Author(s):  
G. Sosinsky ◽  
R. Schekman ◽  
R. Glaeser
Keyword(s):  
Electron Microscopy ◽  
Cell Wall ◽  
Plasma Membrane ◽  
High Resolution Electron Microscopy ◽  
Yeast Cells ◽  
Physiological Conditions ◽  
Resolution Electron ◽  
Yeast Protoplasts ◽  
Intramembraneous Particles ◽  
Crystalline Array

The crystalline patches of intramembraneous particles that form in the yeast plasma membrane, under stationary state physiological conditions, represent a potentially interesting specimen for high resolution electron microscopy. Isolation of these crystalline membrane patches first requires removal of the cell wall and the formation of osmotically fragile yeast protoplasts. In developing a procedure for the isolation of these crystalline membrane patches, we have found that the intramembraneous particles form much larger crystalline patches in protoplasts than in intact yeast cells. We have performed deep etch experiments and have found that the crystalline array of particles is not expressed on the extracellular surface of the plasma membrane.

scholarly journals Sr1/2Ce5/14□1/7WO4: a new modulated ternary scheelite compound

2017 ◽  
Vol 73 (3) ◽  
pp. 466-473 ◽  
Author(s):  
Rafael Hernandez Damascena dos Passos ◽  
Madjid Arab ◽  
Carlson Pereira de Souza ◽  
Christine Leroux
Keyword(s):  
Band Gap ◽  
High Resolution Electron Microscopy ◽  
Partial Ordering ◽  
Promising Candidate ◽  
Atomic Displacements ◽  
Violet Luminescence ◽  
Resolution Electron ◽  
20 Nm ◽  
Cation Sites ◽  
First Time

For the first time, a ternary tetragonal scheelite structure tungstate with strontium and cerium cations, (Sr,Ce)WO4, was synthesized. As much as 35% Ce could be inserted into the structure, leaving 1\over 7 of the (Sr,Ce) cation sites vacant. Partial ordering of Sr and Ce, with atomic displacements, were shown by high-resolution electron microscopy. Two-dimensional incommensurate modulations occur in this material, in small domains 20 nm in size. The band gap of this compound is significantly lower than the band gap of SrWO4and this was related to the distortions of WO4and (Sr,Ce)O8polyhedra. The band gap value of 3.2 eV makes Sr1/2Ce5/14□1/7WO4a promising candidate for violet luminescence.

scholarly journals Intracellular Retention of Glycosylphosphatidyl Inositol-Linked Proteins in Caveolin-Deficient Cells

2002 ◽  
Vol 22 (11) ◽  
pp. 3905-3926 ◽  
Author(s):  
Federica Sotgia ◽  
Babak Razani ◽  
Gloria Bonuccelli ◽  
William Schubert ◽  
Michela Battista ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Cell Surface ◽  
Lipid Rafts ◽  
Recombinant Expression ◽  
Surface Expression ◽  
Cell Surface Expression ◽  
Null Mouse ◽  
Tissue Samples ◽  
Caveolin 1 ◽  
Glycosylphosphatidyl Inositol

ABSTRACT The relationship between glycosylphosphatidyl inositol (GPI)-linked proteins and caveolins remains controversial. Here, we derived fibroblasts from Cav-1 null mouse embryos to study the behavior of GPI-linked proteins in the absence of caveolins. These cells lack morphological caveolae, do not express caveolin-1, and show a ∼95% down-regulation in caveolin-2 expression; these cells also do not express caveolin-3, a muscle-specific caveolin family member. As such, these caveolin-deficient cells represent an ideal tool to study the role of caveolins in GPI-linked protein sorting. We show that in Cav-1 null cells GPI-linked proteins are preferentially retained in an intracellular compartment that we identify as the Golgi complex. This intracellular pool of GPI-linked proteins is not degraded and remains associated with intracellular lipid rafts as judged by its Triton insolubility. In contrast, GPI-linked proteins are transported to the plasma membrane in wild-type cells, as expected. Furthermore, recombinant expression of caveolin-1 or caveolin-3, but not caveolin-2, in Cav-1 null cells complements this phenotype and restores the cell surface expression of GPI-linked proteins. This is perhaps surprising, as GPI-linked proteins are confined to the exoplasmic leaflet of the membrane, while caveolins are cytoplasmically oriented membrane proteins. As caveolin-1 normally undergoes palmitoylation on three cysteine residues (133, 143, and 156), we speculated that palmitoylation might mechanistically couple caveolin-1 to GPI-linked proteins. In support of this hypothesis, we show that palmitoylation of caveolin-1 on residues 143 and 156, but not residue 133, is required to restore cell surface expression of GPI-linked proteins in this complementation assay. We also show that another lipid raft-associated protein, c-Src, is retained intracellularly in Cav-1 null cells. Thus, Golgi-associated caveolins and caveola-like vesicles could represent part of the transport machinery that is necessary for efficiently moving lipid rafts and their associated proteins from the trans-Golgi to the plasma membrane. In further support of these findings, GPI-linked proteins were also retained intracellularly in tissue samples derived from Cav-1 null mice (i.e., lung endothelial and renal epithelial cells) and Cav-3 null mice (skeletal muscle fibers).

scholarly journals Regulation of caveolin-1 membrane trafficking by the Na/K-ATPase

2008 ◽  
Vol 182 (6) ◽  
pp. 1153-1169 ◽  
Author(s):  
Ting Cai ◽  
Haojie Wang ◽  
Yiliang Chen ◽  
Lijun Liu ◽  
William T Gunning ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Cell Surface ◽  
Long Range ◽  
Membrane Trafficking ◽  
Endocytic Trafficking ◽  
Binding Motif ◽  
Caveolin 1 ◽  
Pumping Function ◽  
Α1 Subunit

Here, we show that the Na/K-ATPase interacts with caveolin-1 (Cav1) and regulates Cav1 trafficking. Graded knockdown of Na/K-ATPase decreases the plasma membrane pool of Cav1, which results in a significant reduction in the number of caveolae on the cell surface. These effects are independent of the pumping function of Na/K-ATPase, and instead depend on interaction between Na/K-ATPase and Cav1 mediated by an N-terminal caveolin-binding motif within the ATPase α1 subunit. Moreover, knockdown of the Na/K-ATPase increases basal levels of active Src and stimulates endocytosis of Cav1 from the plasma membrane. Microtubule-dependent long-range directional trafficking in Na/K-ATPase–depleted cells results in perinuclear accumulation of Cav1-positive vesicles. Finally, Na/K-ATPase knockdown has no effect on processing or exit of Cav1 from the Golgi. Thus, the Na/K-ATPase regulates Cav1 endocytic trafficking and stabilizes the Cav1 plasma membrane pool.

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