Identification and characterization of Saccharomyces cerevisiae dihydrosphingosine-1-phosphate phosphatase.
AuthorsMao C, Wadleigh M, Jenkins GM, Hannun YA, Obeid LM
JournalJ Biol Chem
PubMed ID9353337
We have identified the yeast sphingosine resistance gene (YSR2) of Saccharomyces cerevisiae as encoding a protein that specifically dephosphorylates dihydrosphingosine 1-phosphate (DHS-1-P), and we refer to this protein as dihydrosphingosine-1-phosphate phosphatase. Overexpression of YSR2 conferred sphingosine resistance to the dihydrosphingosine-1-P lyase-defective mutant (JS16) of S. cerevisiae, which is hypersensitive to ... More
Determination of the intracellular sites and topology of glucosylceramide synthesis in rat liver.
AuthorsFuterman AH, Pagano RE
JournalBiochem J
PubMed ID1747103
We examined the intracellular site(s) and topology of glucosylceramide (GlcCer) synthesis in subcellular fractions from rat liver, using radioactive and fluorescent ceramide analogues as precursors, and compared these results with those obtained in our recent study of sphingomyelin (SM) synthesis in rat liver [Futerman, Stieger, Hubbard & Pagano (1990) J. ... More
AuthorsChiang AS, Lin WY, Liu HP, Pszczolkowski MA, Fu TF, Chiu SL, Holbrook GL
JournalProc Natl Acad Sci U S A
PubMed ID11773617
'In vertebrates, the N-methyl-D-aspartate subtype of glutamate receptors (NMDAR) appears to play a role in neuronal development, synaptic plasticity, memory formation, and pituitary activity. However, functional NMDAR have not yet been characterized in insects. We have now demonstrated immunohistochemically glutamatergic nerve terminals in the corpora allata of an adult female ... More
An endocytosed TGN38 chimeric protein is delivered to the TGN after trafficking through the endocytic recycling compartment in CHO cells.
AuthorsGhosh RN, Mallet WG, Soe TT, McGraw TE, Maxfield FR
JournalJ Cell Biol
PubMed ID9722606
'To examine TGN38 trafficking from the cell surface to the TGN, CHO cells were stably transfected with a chimeric transmembrane protein, TacTGN38. We used fluorescent and 125I-labeled anti-Tac IgG and Fab fragments to follow TacTGN38''s postendocytic trafficking. At steady-state, anti-Tac was mainly in the TGN, but shortly after endocytosis it ... More
The signal anchor and stem regions of the beta-galactoside alpha 2,6-sialyltransferase may each act to localize the enzyme to the Golgi apparatus.
AuthorsColley KJ, Lee EU, Paulson JC
JournalJ Biol Chem
PubMed ID1560012
'The beta-galactoside alpha 2,6-sialyltransferase has been localized to the trans cisternae of the Golgi apparatus and the trans Golgi network where it transfers sialic acid residues to terminal positions on N-linked oligosaccharides. It is a type II transmembrane protein possessing a 9-amino acid amino-terminal cytoplasmic tail, a 17-amino acid signal ... More
Kinesin-mediated organelle translocation revealed by specific cellular manipulations.
AuthorsFeiguin F, Ferreira A, Kosik KS, Caceres A
JournalJ Cell Biol
PubMed ID7962067
'The distribution of membrane-bound organelles was studied in cultured hippocampal neurons after antisense oligonucleotide suppression of the kinesin-heavy chain (KHC). We observed reduced 3,3''-dihexyloxacarbocyanine iodide (DiOC6(3)) fluorescent staining in neurites and growth cones. In astrocytes, KHC suppression results in the disappearance of the DiOC6(3)-positive reticular network from the cell periphery, ... More
Forskolin inhibits and reverses the effects of brefeldin A on Golgi morphology by a cAMP-independent mechanism.
'Brefeldin A (BFA) causes rapid redistribution of Golgi proteins into the ER, leaving no definable Golgi apparatus, and blocks transport of proteins into post-Golgi compartments in the cell. In this study we follow the disassembly of the Golgi apparatus in BFA-treated, living cells labeled with NBD-ceramide and demonstrate that forskolin ... More
Analysis of P-glycoprotein-mediated membrane transport in human peripheral blood lymphocytes using the UIC2 shift assay.
AuthorsPark SW, Lomri N, Simeoni LA, Fruehauf JP, Mechetner E
JournalCytometry A
PubMed ID12766968
'BACKGROUND: During transport-associated adenosine triphosphate hydrolysis, P-glycoprotein (Pgp) undergoes conformation transitions detected by UIC2, a functional anti-Pgp monoclonal antibody. A newly developed UIC2 shift assay is based on increased UIC2 reactivity in the presence of Pgp substrates. All peripheral blood leukocytes express low Pgp levels. The existing antibody-based detection methods ... More
Coupling assembly of the E-cadherin/beta-catenin complex to efficient endoplasmic reticulum exit and basal-lateral membrane targeting of E-cadherin in polarized MDCK cells.
AuthorsChen YT, Stewart DB, Nelson WJ
JournalJ Cell Biol
PubMed ID10037790
'The E-cadherin/catenin complex regulates Ca++-dependent cell-cell adhesion and is localized to the basal-lateral membrane of polarized epithelial cells. Little is known about mechanisms of complex assembly or intracellular trafficking, or how these processes might ultimately regulate adhesion functions of the complex at the cell surface. The cytoplasmic domain of E-cadherin ... More
Differential targeting of glucosylceramide and galactosylceramide analogues after synthesis but not during transcytosis in Madin-Darby canine kidney cells.
Authorsvan Genderen I, van Meer G
JournalJ Cell Biol
PubMed ID7593186
'A short-chain analogue of galactosylceramide (6-NBD-amino-hexanoyl-galactosylceramide, C6-NBD-GalCer) was inserted into the apical or the basolateral surface of MDCK cells and transcytosis was monitored by depleting the opposite cell surface of the analogue with serum albumin. In MDCK I cells 32% of the analogue from the apical surface and 9% of ... More
Lipid metabolism in Chlamydia trachomatis-infected cells: directed trafficking of Golgi-derived sphingolipids to the chlamydial inclusion.
AuthorsHackstadt T, Scidmore MA, Rockey DD
JournalProc Natl Acad Sci U S A
PubMed ID7761416
'Chlamydia trachomatis undergoes its entire life cycle within an uncharacterized intracellular vesicle that does not fuse with lysosomes. We used a fluorescent Golgi-specific probe, (N-[7-(4-nitrobenzo-2-oxa-1,3-diazole)]) aminocaproylsphingosine (C6-NBD-Cer), in conjunction with conventional fluorescence or confocal microscopy to identify interactions between the Golgi apparatus and the chlamydial inclusion. We observed not only ... More
Differential effects of brefeldin A on transport of secretory and lysosomal proteins.
AuthorsStrous GJ, van Kerkhof P, van Meer G, Rijnboutt S, Stoorvogel W
JournalJ Biol Chem
PubMed ID8428908
'Brefeldin A (BFA) rapidly blocks anterograde exocytotic transport through the Golgi complex. Sustained retrograde traffic induced by brefeldin A causes redistribution of constituents of the Golgi, but not the trans-Golgi network (TGN), to the endoplasmic reticulum (ER). In the present study on HepG2 cells, we have observed a differential effect ... More
Energy-dependent accumulation of daunorubicin into subcellular compartments of human leukemia cells and cytoplasts.
AuthorsSlapak CA, Lecerf JM, Daniel JC, Levy SB
JournalJ Biol Chem
PubMed ID1350280
'Anthracycline accumulation was evaluated by flow cytometry or radiolabeled drug assays in cells and cytoplasts (enucleated cells) prepared from parental and multidrug-resistant human K562 leukemia cells. Treatment with energy inhibitors, such as dinitrophenol (DNP) or sodium azide/deoxyglucose, led to a marked decrease in daunorubicin accumulation in parental cells and cytoplasts. ... More
The effect of apical and basolateral lipids on the function of the band 3 anion exchange protein.
Authorsvan't Hof W, Malik A, Vijayakumar S, Qiao J, van Adelsberg J, Al-Awqati Q
JournalJ Cell Biol
PubMed ID9362512
'Although many polarized proteins are sorted to the same membrane domain in all epithelial tissues, there are some that exhibit a cell type-specific polarity. We recently found that band 3 (the anion exchanger AE1) was present in the apical membrane of a renal intercalated cell line when these cells were ... More
Sphingolipids and glycoproteins are differentially trafficked to the Chlamydia trachomatis inclusion.
AuthorsScidmore MA, Fischer ER, Hackstadt T
JournalJ Cell Biol
PubMed ID8707822
'Chlamydia trachomatis is an obligate intracellular pathogen that multiples within the confines of a membrane-bound vacuole called an inclusion. Approximately 40-50% of the sphingomyelin synthesized from exogenously added NBD-ceramide is specifically transported from the Golgi apparatus to the chlamydial inclusion (Hackstadt, T., M.A. Scidmore, and D.D. Rockey. 1995. Proc. Natl. ... More
Determination of intracellular organelles implicated in daunorubicin cytoplasmic sequestration in multidrug-resistant MCF-7 cells using fluorescence microscopy image analysis.
AuthorsBour-Dill C, Gramain MP, Merlin JL, Marchal S, Guillemin F
JournalCytometry
PubMed ID10655559
'BACKGROUND: Anthracycline resistance is known to be mediated by P-glycoprotein (P-gp) or multidrug-resistance related protein (MRP) as well as intracellular sequestration of drugs. METHODS: The resistance phenotype of doxorubicin-selected MCF-7(DXR) human breast adenocarcinoma cell line was characterized by cellular and nuclear daunorubicin efflux, P-gp and MRP expression and apoptosis induction. ... More
Tubulovesicular processes emerge from trans-Golgi cisternae, extend along microtubules, and interlink adjacent trans-golgi elements into a reticulum.
AuthorsCooper MS, Cornell-Bell AH, Chernjavsky A, Dani JW, Smith SJ
JournalCell
PubMed ID2180583
'Morphological dynamics and membrane transport within the living Golgi apparatus of astrocytes labeled with NBD-ceramide were imaged using both electronically enhanced fluorescence video and laser confocal microscopy. In time-lapse recordings, continuous tubulovesicular processes are observed to emerge from trans-Golgi elements and extend along microtubules at average rates of 0.4 microns/s. ... More
A vital stain for the Golgi apparatus.
AuthorsLipsky NG, Pagano RE
JournalScience
PubMed ID2581316
'The Golgi complex, a membranous organelle with important functions in membrane traffic and macromolecular synthesis, has been stained in living cells with a fluorescent sphingolipid. Cells were first incubated with liposomes containing N-[7-(4-nitrobenzo-2-oxa-1,3-diazole)]-6-aminocaproyl sphingosine (C6-NBD-ceramide), or with a bovine serum albumin complex of the fluorescent lipid, and then examined by ... More
Plasmodium falciparum exports the Golgi marker sphingomyelin synthase into a tubovesicular network in the cytoplasm of mature erythrocytes.
AuthorsElmendorf HG, Haldar K
JournalJ Cell Biol
PubMed ID8106545
'This work describes two unusual features of membrane development in a eukaryotic cell. (a) The induction of an extensive network of tubovesicular membranes by the malaria parasite Plasmodium falciparum in the cytoplasm of the mature erythrocyte, and its visualization with two ceramide analogues C5-DMB-ceramide and C6-NBD-ceramide. "Sectioning" of the infected ... More
Tumor necrosis factor induces ceramide oscillations and negatively controls sphingolipid synthases by caspases in apoptotic Kym-1 cells.
AuthorsBourteele S, Hausser A, Döppler H, Horn-Müller J, Röpke C, Schwarzmann G, Pfizenmaier K, Müller G
JournalJ Biol Chem
PubMed ID9813032
'The role, origin, and mode of action of the lipid messenger ceramide in programmed cell death and its linkage to receptor-associated apoptotic signal proteins is still unresolved. We show here in Kym-1 rhabdomyosarcoma cells that tumor necrosis factor (TNF)-induced apoptosis is preceded by a multiphasic increase in intracellular ceramide levels. ... More
Cholesterol deprivation affects the fluorescence properties of a ceramide analog at the Golgi apparatus of living cells.
AuthorsMartin OC, Comly ME, Blanchette-Mackie EJ, Pentchev PG, Pagano RE
JournalProc Natl Acad Sci U S A
PubMed ID8464873
'Previous studies have established that a fluorescent analog of ceramide, N-[7-(4-nitrobenzo-2-oxa-1,3-diazole)] -6-aminohexanoyl-D-erythro-sphingosine (C6-NBD-Cer), is a vital stain for the Golgi apparatus and a useful tool for studying the sorting and transport of sphingolipids along the secretory pathway in animal cells. Here, we examine the effects of various culture conditions on ... More
Chlamydia trachomatis interrupts an exocytic pathway to acquire endogenously synthesized sphingomyelin in transit from the Golgi apparatus to the plasma membrane.
AuthorsHackstadt T, Rockey DD, Heinzen RA, Scidmore MA
JournalEMBO J
PubMed ID8605892
'Chlamydia trachomatis acquires C6-NBD-sphingomyelin endogenously synthesized from C6-NBD-ceramide and transported to the vesicle (inclusion) in which they multiply. Here we explore the mechanisms of this unusual trafficking and further characterize the association of the chlamydial inclusion with the Golgi apparatus. Endocytosed chlamydiae are trafficked to the Golgi region and begin ... More
Restricted fusion of Chlamydia trachomatis vesicles with endocytic compartments during the initial stages of infection.
AuthorsScidmore MA, Fischer ER, Hackstadt T
JournalInfect Immun
PubMed ID12540580
'The chlamydial inclusion occupies a unique niche within the eukaryotic cell that does not interact with endocytic compartments but instead is fusogenic with a subset of sphingomyelin-containing exocytic vesicles. The Chlamydia trachomatis inclusion acquires these distinctive properties by as early as 2 h postinfection as demonstrated by the ability to ... More
Penetration of Toxoplasma gondii into host cells induces changes in the distribution of the mitochondria and the endoplasmic reticulum.
Authorsde Melo EJ, de Carvalho TU, de Souza W
JournalCell Struct Funct
PubMed ID1473161
'Fluorescence microscopy, using dyes which specifically label mitochondria, endoplasmic reticulum and the Golgi complex, and transmission electron microscopy, were used to analyze the changes which occur in the organization of these structures during interaction of Toxoplasma gondii with host cells. In uninfected cells the mitochondria are long filamentous structures which ... More
An endoplasmic reticulum retention signal in the CD3 epsilon chain of the T-cell receptor.
AuthorsMallabiabarrena A, Fresno M, Alarcón B
JournalNature
PubMed ID1535117
'Isolated polypeptide chains of the T-cell antigen receptor complex are degraded or retained in the endoplasmic reticulum (ER). Assembly of the multisubunit complex allows the individual chains to escape retention in the ER and to be expressed on the cell surface. We engineered a series of deletions in the CD3 ... More
Complex sphingolipid synthesis in plants: characterization of inositolphosphorylceramide synthase activity in bean microsomes.
AuthorsBromley PE, Li YO, Murphy SM, Sumner CM, Lynch DV
JournalArch Biochem Biophys
PubMed ID12941304
'Complex glycophosphosphingolipids present in plants are composed of ceramide, inositolphosphate, and diverse polar oligosaccharide substituents. The activity of inositolphosphorylceramide (IPC) synthase (phosphatidylinositol:ceramide inositolphosphate transferase), the enzyme proposed to catalyze the initial committed step in the formation of these complex sphingolipids, was characterized in wax bean hypocotyl microsomes. Enzyme activity was ... More
The spatial organization of corneal endothelial cytoskeletal proteins and their relationship to the apical junctional complex.
'PURPOSE. To determine the spatial organization of the major cytoskeletal proteins and their relationship to the apical junctional complex (AJC) in the normal rabbit corneal endothelium. METHODS. Normal endothelial cytoskeletal structure in three dimensions was studied in rabbit eyes by laser scanning confocal microscopy after en bloc immunocytochemical staining of ... More
Mutant Rab7 causes the accumulation of cathepsin D and cation-independent mannose 6-phosphate receptor in an early endocytic compartment.
AuthorsPress B, Feng Y, Hoflack B, Wandinger-Ness A
JournalJ Cell Biol
PubMed ID9490721
'Stable BHK cell lines inducibly expressing wild-type or dominant negative mutant forms of the rab7 GTPase were isolated and used to analyze the role of a rab7-regulated pathway in lysosome biogenesis. Expression of mutant rab7N125I protein induced a dramatic redistribution of cation-independent mannose 6-phosphate receptor (CI-MPR) from its normal perinuclear ... More
Reconstitution of constitutive secretion using semi-intact cells: regulation by GTP but not calcium.
AuthorsMiller SG, Moore HP
JournalJ Cell Biol
PubMed ID1986006
'Regulated exocytosis in many permeabilized cells can be triggered by calcium and nonhydrolyzable GTP analogues. Here we examine the role of these effectors in exocytosis of constitutive vesicles using a system that reconstitutes transport between the trans-Golgi region and the plasma membrane. Transport is assayed by two independent methods: the ... More
Intracellular targeting and trafficking of thrombin receptors. A novel mechanism for resensitization of a G protein-coupled receptor.
AuthorsHein L, Ishii K, Coughlin SR, Kobilka BK
JournalJ Biol Chem
PubMed ID7961693
'The receptor for the protease thrombin is a member of the G protein-coupled receptor family, but is activated by a unique proteolytic mechanism. The irreversibility of this proteolytic mechanism and the fact that the ligand is tethered to its receptor raise special questions about inactivation of cleaved receptors and recovery ... More
Association of cytoplasmic free Ca2+ gradients with subcellular organelles.
AuthorsWahl M, Sleight RG, Gruenstein E
JournalJ Cell Physiol
PubMed ID1537888
'Previous investigations have identified gradients of intracellular free (Ca2+)i (Ca2+i) in the cytoplasm of human fibroblasts. In this study we have compared the spatial distribution of these gradients with the subcellular distribution of cytoplasmic organelles. Using the Ca(2+)-sensitive dye fura-2 and organelle-specific fluorescent dyes, we have found that the highest ... More
Molecular trapping of a fluorescent ceramide analogue at the Golgi apparatus of fixed cells: interaction with endogenous lipids provides a trans-Golgi marker for both light and electron microscopy.
AuthorsPagano RE, Sepanski MA, Martin OC
JournalJ Cell Biol
PubMed ID2478562
'We have previously shown that a fluorescent derivative of ceramide, N-(epsilon-7-nitrobenz-2-oxa-1,3-diazol-4-yl-aminocaproyl)-D-eryth ro-sphingosin e (C6-NBD-Cer), vitally stains the Golgi apparatus of cells (Lipsky, N. G., and R. E. Pagano. 1985. Science (Wash. DC). 228:745-747). In the present paper we demonstrate that C6-NBD-Cer also accumulates at the Golgi apparatus of fixed cells ... More
Induction of ceramide glucosyltransferase activity in cultured human keratinocytes. Correlation with culture differentiation.
AuthorsSando GN, Howard EJ, Madison KC
JournalJ Biol Chem
PubMed ID8703011
'Ceramides are the major component of the extracellular lipids that comprise the epidermal permeability barrier. They are derived from glucosylceramides (GlcCer) upon their extrusion from lamellar granules into the extracellular space in the upper layers of the epidermis. To better understand the regulation of the unique pathway for ceramide production ... More
A non-exchangeable fluorescent phospholipid analog as a membrane traffic marker of the endocytic pathway.
AuthorsWillem J, ter Beest M, Scherphof G, Hoekstra D
JournalEur J Cell Biol
PubMed ID2076704
'The fluorescent phospholipid analog N-(lissamine rhodamine B sulfonyl)phosphatidylethanolamine (N-Rh-PE) was inserted into the plasma membrane of Baby hamster kidney cells at low temperature (2 degrees C). The mobility characteristics of the analog--as revealed by fluorescence photobleaching recovery--were very similar to those of membrane-inserted 1-acyl-2[6-[N-(7-nitro-2,1,3-benzoxadiazol-4-yl) amino]caproyl] phosphatidylcholine (C6-NBD-PC). Upon warming to ... More
Normal- and reverse-phase HPLC separations of fluorescent (NBD) lipids.
AuthorsMartin OC, Pagano RE
JournalAnal Biochem
PubMed ID3812987
'We have developed two high-performance liquid chromatography methods for separating a number of fluorescent 4-nitrobenzo-2-oxa-1,3-diazole (NBD) analogs of glycerolipids and sphingolipids. Samples of fluorescent lipid analogs containing NBD-aminocaproyl (C6-NBD) or NBD-aminododecanoyl (C12-NBD) acyl chains were synthesized and analyzed by the following HPLC methods. An isocratic normal-phase method permitted resolution of ... More
Ceramide inhibits axonal growth and nerve growth factor uptake without compromising the viability of sympathetic neurons.
Authorsde Chaves EP, Bussiere M, MacInnis B, Vance DE, Campenot RB, Vance JE
JournalJ Biol Chem
PubMed ID11454862
'Ceramide inhibits axonal growth of cultured rat sympathetic neurons when the ceramide content of distal axons, but not cell bodies, is increased (Posse de Chaves, E. I., Bussiere, M. Vance, D. E., Campenot, R. B., and Vance, J.E. (1997) J. Biol. Chem. 272, 3028-3035). We now report that inhibition of ... More
Role of microtubules in transferrin receptor transport from the cell surface to endosomes and the Golgi complex.
AuthorsJin M, Snider MD
JournalJ Biol Chem
PubMed ID8349714
'Transferrin receptor (TfR) follows complex pathways of transport after endocytosis from the cell surface. Most TfR is transported to endosomes and returns rapidly to the cell surface. In addition, approximately 10% of the internalized receptor recycles through the Golgi complex. To examine the role of microtubules in TfR traffic, K562 ... More
Developmental induction of Golgi structure and function in the primitive eukaryote Giardia lamblia.
AuthorsLuján HD, Marotta A, Mowatt MR, Sciaky N, Lippincott-Schwartz J, Nash TE
JournalJ Biol Chem
PubMed ID7876232
'A fundamental characteristic of eukaryotic cells is the presence of membrane-bound compartments and membrane transport pathways in which the Golgi complex plays a central role in the selective processing, sorting, and secretion of proteins. The parasitic protozoan Giardia lamblia belongs to the earliest identified lineage among eukaryotes and therefore offers ... More
Analysis of lipoproteins by capillary zone electrophoresis in microfluidic devices: assay development and surface roughness measurements.
AuthorsWeiller BH, Ceriotti L, Shibata T, Rein D, Roberts MA, Lichtenberg J, German JB, de Rooij NF, Verpoorte E
JournalAnal Chem
PubMed ID12033263
'The development of a new assay for lipoproteins by capillary electrophoresis in fused-silica capillaries and in glass microdevices is described in this paper. The separation of low-density (LDL) and high-density (HDL) lipoproteins by capillary zone electrophoresis is demonstrated in fused-silica capillaries with both UV absorption and laser-induced fluorescence detection. This ... More
Hierarchy of mechanisms involved in generating Na/K-ATPase polarity in MDCK epithelial cells.
AuthorsMays RW, Siemers KA, Fritz BA, Lowe AW, van Meer G, Nelson WJ
JournalJ Cell Biol
PubMed ID7657695
'We have studied mechanisms involved in generating a polarized distribution of Na/K-ATPase in the basal-lateral membrane of two clones of MDCK II cells. Both clones exhibit polarized distributions of marker proteins of the apical and basal-lateral membranes, including Na/K-ATPase, at steady state. Newly synthesized Na/K-ATPase, however, is delivered from the ... More
Vasodilation by the calcium-mobilizing messenger cyclic ADP-ribose.
AuthorsBoittin FX, Dipp M, Kinnear NP, Galione A, Evans AM
JournalJ Biol Chem
PubMed ID12486132
'In artery smooth muscle, adenylyl cyclase-coupled receptors such as beta-adrenoceptors evoke Ca(2+) signals, which open Ca(2+)-activated potassium (BK(Ca)) channels in the plasma membrane. Thus, blood pressure may be lowered, in part, through vasodilation due to membrane hyperpolarization. The Ca(2+) signal is evoked via ryanodine receptors (RyRs) in sarcoplasmic reticulum proximal ... More
Chicken erythroid AE1 anion exchangers associate with the cytoskeleton during recycling to the Golgi.
AuthorsGhosh S, Cox KH, Cox JV
JournalMol Biol Cell
PubMed ID9950688
'Chicken erythroid AE1 anion exchangers receive endoglycosidase F (endo F)-sensitive sugar modifications in their initial transit through the secretory pathway. After delivery to the plasma membrane, anion exchangers are internalized and recycled to the Golgi where they acquire additional N-linked modifications that are resistant to endo F. During recycling, some ... More
Fluorescent lipid uptake and transport in adult Schistosoma mansoni.
AuthorsMoffat D, Kusel JR
JournalParasitology
PubMed ID1437279
'Fluorescent lipophilic compounds can be used to label the surface membrane of Schistosoma mansoni by adding the compound in small amounts of organic solvents to aqueous medium in vitro. Under these conditions it is difficult to follow routes of distribution of the label. Here we have absorbed nitrobenzoxadiazolamine methylamino-(NBD)-ceramides to ... More
Reconstitution of ATP- and cytosol-dependent transport of de novo synthesized ceramide to the site of sphingomyelin synthesis in semi-intact cells.
AuthorsFunakoshi T, Yasuda S, Fukasawa M, Nishijima M, Hanada K
JournalJ Biol Chem
PubMed ID10882735
'Transport of ceramide synthesized at the endoplasmic reticulum to the Golgi compartment, where sphingomyelin (SM) synthase exists, was reconstituted within semi-intact Chinese hamster ovary cells. When [(3)H]ceramide that had been produced from [(3)H]sphingosine at 15 degrees C in perforated cells was chased at 37 degrees C, [(3)H]ceramide-to-[(3)H]SM conversion occurred in ... More
Serial quantitative image analysis and confocal microscopy of hepatic uptake, intracellular distribution and biliary secretion of a fluorescent bile acid analog in rat hepatocyte doublets.
AuthorsKitamura T, Gatmaitan Z, Arias IM
JournalHepatology
PubMed ID2124205
'To characterize the poorly understood mechanisms of intracellular transport of bile acids, fluorescein isothiocyanate-glycocholate was synthesized and its ring-OH-linked structure established by fast atom bombardment, mass spectroscopy and 13C nuclear magnetic resonance. Biliary secretion of fluorescein isothiocyanate-glycocholate and [14C]-labeled glycocholate in rats was similar, in contrast to the biliary secretion ... More
Visualization and functional analysis of RNA-dependent RNA polymerase lattices.
AuthorsLyle JM, Bullitt E, Bienz K, Kirkegaard K
JournalScience
PubMed ID12077417
'Positive-strand RNA viruses such as poliovirus replicate their genomes on intracellular membranes of their eukaryotic hosts. Electron microscopy has revealed that purified poliovirus RNA-dependent RNA polymerase forms planar and tubular oligomeric arrays. The structural integrity of these arrays correlates with cooperative RNA binding and RNA elongation and is sensitive to ... More
The End2 mutation in CHO cells slows the exit of transferrin receptors from the recycling compartment but bulk membrane recycling is unaffected.
AuthorsPresley JF, Mayor S, Dunn KW, Johnson LS, McGraw TE, Maxfield FR
JournalJ Cell Biol
PubMed ID8376460
'We have characterized a new CHO cell line (12-4) derived from a parental line, TRVb-1, that expresses the human transferrin receptor. This mutant belongs to the end2 complementation group of endocytosis mutants. Like other end2 mutants, the endosomes in 12-4 cells show a partial acidification defect. These cells internalize LDL ... More
The effect of Chlamydia trachomatis infection on the host cell cytoskeleton and membrane compartments.
AuthorsCampbell S, Richmond SJ, Yates PS
JournalJ Gen Microbiol
PubMed ID2483409
'Human epithelial cells and the McCoy cell line were infected with Chlamydia trachomatis, serotype E. The organization of the cytoplasm was then studied with probes which stained cytoskeletal components and membrane compartments. The major actin-containing stress fibre bundles were not associated with inclusions due to the peri-basal and peri-apical location ... More
Endoplasmic reticulum and Golgi-like elements in Entamoeba.
AuthorsMazzuco A, Benchimol M, De Souza W
JournalMicron
PubMed ID9377074
'The cytoplasm of Entamoeba is characterized by the presence of a large number of vesicles of different size and shape. Previous electron microscopic studies have not clearly revealed the presence of the endoplasmic reticulum and the Golgi complex. In the present study two approaches were used aimed at the identification ... More
Pathway of C6-NBD-Ceramide on the host cell infected with Toxoplasma gondii.
Authorsde Melo EJ, de Souza W
JournalCell Struct Funct
PubMed ID8726473
'Fluorescence microscopy, with dyes analog of ceramide, and transmission electron microscopy, were used to analyze lipid traffic during interaction of Toxoplasma gondii with host cells. It is C6-NBD-Ceramide (C6-NBD-Cer), a fluorescent analog of ceramide, stained the Golgi complex where was metabolized into fluorescent sphingolipid and glucosylceramide, and translocated via the ... More
Inhibition of sphingolipid synthesis affects axonal outgrowth in cultured hippocampal neurons.
AuthorsHarel R, Futerman AH
JournalJ Biol Chem
PubMed ID8314804
'Neuronal growth is regulated by both extracellular and cellular determinants and is believed to proceed by the addition of new membrane material at the growth cone. To determine whether lipid synthesis is necessary to maintain neuronal growth, we have examined the effect of Fumonisin B1, an inhibitor of ceramide synthesis, ... More
DS28-6, a temperature-sensitive mutant of Chinese hamster ovary cells, expresses key phenotypic changes associated with brefeldin A treatment.
AuthorsZuber C, Roth J, Misteli T, Nakano A, Moremen K
JournalProc Natl Acad Sci U S A
PubMed ID1946407
'The temperature-sensitive Chinese hamster ovary (CHO) cell mutant DS28-6 has been previously shown to be pleiotropically defective in protein secretion. We have examined the mutant cells to determine the intracellular site of the block in secretion. By transmission electron microscopy a time-dependent disassembly of the Golgi apparatus was found under ... More
Temporal analysis of the developing Chlamydia psittaci inclusion by use of fluorescence and electron microscopy.
AuthorsRockey DD, Fischer ER, Hackstadt T
JournalInfect Immun
PubMed ID8926099
'The chlamydiae are obligate intracellular parasites that develop and multiply within a vacuole (termed an inclusion) that does not fuse with lysosomes. Inclusion morphology varies dramatically among the different chlamydiae, particularly within the species Chlamydia psittaci. Some strains develop within a single vacuole, while the mature inclusion of other strains ... More
Lipopolysaccharide-induced ischemic tolerance is associated with increased levels of ceramide in brain and in plasma.
AuthorsZimmermann C, Ginis I, Furuya K, Klimanis D, Ruetzler C, Spatz M, Hallenbeck JM
JournalBrain Res
PubMed ID11259760
'Intravenous administration of lipopolysaccharide (LPS) (0.9 mg/kg) has been shown to induce ischemic tolerance in spontaneously hypertensive rats (SHR). TNF-alpha is believed to play a crucial role in preconditioning as its inhibition with TNF-alpha-binding protein abolished tolerance. Our recent studies (Liu et al., Am. J. Physiol. 278 C144, 2000) have ... More
Sphingolipid metabolism in cultured fibroblasts: microscopic and biochemical studies employing a fluorescent ceramide analogue.
AuthorsLipsky NG, Pagano RE
JournalProc Natl Acad Sci U S A
PubMed ID6573674
'A fluorescent analogue of ceramide, N-[7-(4-nitrobenzo-2-oxa-1,3-diazole)]-epsilon-aminocaproyl sphingosine (C6-NBD-ceramide), was used to investigate sphingolipid metabolism in Chinese hamster fibroblasts. C6-NBD-ceramide was incorporated into small unilamellar dioleoyl phosphatidylcholine vesicles and incubated with cells in monolayer culture at 2 degrees C, resulting in rapid and preferential transfer of the labeled ceramide from vesicles ... More
Induction of endocytic vesicles by exogenous C(6)-ceramide.
AuthorsLi R, Blanchette-Mackie EJ, Ladisch S
JournalJ Biol Chem
PubMed ID10409665
'Ceramide is a newly discovered second messenger that has been shown to cause cell growth arrest and apoptosis. Here, we present evidence that exogenously added C(6)-ceramide induces enlargement of late endosomes and lysosomes. 10 microM C(6)-ceramide caused the formation of numerous vesicles of varying sizes (2-10 micrometers) in fibroblasts (3T3-L1 ... More
Inhibition of glycoprotein traffic through the secretory pathway by ceramide.
AuthorsRosenwald AG, Pagano RE
JournalJ Biol Chem
PubMed ID8383117
'Incubation of vesicular stomatitis virus-infected cells with short-chain, cell-permeable ceramide (Cer) analogs decreased the rate of viral glycoprotein transport through the Golgi complex and reduced the number of infectious virions released from cells in a concentration-dependent manner. These effects appeared to be caused directly by Cer, rather than by one ... More
Generation of specific deoxynojirimycin-type inhibitors of the non-lysosomal glucosylceramidase.
AuthorsOverkleeft HS, Renkema GH, Neele J, Vianello P, Hung IO, Strijland A, van der Burg AM, Koomen GJ, Pandit UK, Aerts JM
JournalJ Biol Chem
PubMed ID9756888
'The existence of a non-lysosomal glucosylceramidase in human cells has been documented (van Weely, S., Brandsma, M., Strijland, A., Tager, J. M., and Aerts, J. M. F. G. (1993) Biochim. Biophys. Acta 1181, 55-62). Hypothetically, the activity of this enzyme, which is localized near the cell surface, may influence ceramide-mediated ... More
Activation of 5-[125I]iodonaphthyl-1-azide via excitation of fluorescent (N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)) lipid analogs in living cells. A potential tool for identification of compartment-specific proteins and proteins involved in intracellular transport and metabolism of lipids.
AuthorsRosenwald AG, Pagano RE, Raviv Y
JournalJ Biol Chem
PubMed ID2033068
'We describe a new technique for analysis of proteins located near fluorescent lipid analogs in intact living cells using the membrane-permeant, photoactivatable probe, 5-[125I]iodonaphthyl-1-azide ([125I]INA). [125I] INA can be activated directly with UV light or indirectly through excitation of adjacent fluorophores (photosensitizers) with visible light to modify nearby proteins covalently ... More
Transferrin receptor containing the SDYQRL motif of TGN38 causes a reorganization of the recycling compartment but is not targeted to the TGN.
AuthorsJohnson AO, Ghosh RN, Dunn KW, Garippa R, Park J, Mayor S, Maxfield FR, McGraw TE
JournalJ Cell Biol
PubMed ID8991088
'The SDYQRL motif of the cytoplasmic domain of TGN38 is involved in targeting TGN38 from endosomes to the TGN. To create a system for studying this pathway, we replaced the native transferrin receptor (TR) internalization motif (YTRF) with the SDYQRL TGN-targeting motif. The advantages of using TR as a reporter ... More
Involvement of sphingosine in mitochondria-dependent Fas-induced apoptosis of type II Jurkat T cells.
AuthorsCuvillier O, Edsall L, Spiegel S
JournalJ Biol Chem
PubMed ID10747891
'Exposure to anti-Fas antibody in Jurkat cells (type II cells), which are characterized by a weak caspase-8 activation at the death-inducing signaling complex (DISC), induced a biphasic increase in ceramide levels. The early generation of ceramide preceded transient activation of acidic ceramidase and subsequent production of sphingosine, followed by cytochrome ... More
Cloning and characterization of a novel human alkaline ceramidase. A mammalian enzyme that hydrolyzes phytoceramide.
'Ceramidases are enzymes involved in regulating cellular levels of ceramides, sphingoid bases, and their phosphates. Based on sequence homology to the yeast alkaline ceramidases YPC1p (Mao, C., Xu, R., Bielawska, A., and Obeid, L. M. (2000) J. Biol. Chem. 275, 6876--6884) and YDC1p (Mao, C., Xu, R., Bielawska, A., Szulc, ... More
Abnormal processing of Golgi elements and lysosomes in Tangier disease.
AuthorsRobenek H, Schmitz G
JournalArterioscler Thromb
PubMed ID2065025
'It has been demonstrated that the cellular defect in Tangier disease is associated with morphological abnormalities in the lysosomal compartment and in the Golgi apparatus of mononuclear phagocytes (MNPs) after lipid loading by exposure to acetylated low density lipoproteins (acetyl-LDLs). On exposure to acetyl-LDL, Tangier MNPs accumulate two unusual types ... More
D-threo-1-phenyl-2-decanoylamino-3-morpholino-1-propanol alters cellular cholesterol homeostasis by modulating the endosome lipid domains.
AuthorsMakino A, Ishii K, Murate M, Hayakawa T, Suzuki Y, Suzuki M, Ito K, Fujisawa T, Matsuo H, Ishitsuka R, Kobayashi T
JournalBiochemistry
PubMed ID16584188
'D-threo-1-phenyl-2-decanoylamino-3-morpholino-1-propanol (D-PDMP) is a frequently used inhibitor of glycosphingolipid biosynthesis. However, some interesting characteristics of D-PDMP cannot be explained by the inhibition of glycolipid synthesis alone. In the present study, we showed that d-PDMP inhibits the activation of lysosomal acid lipase by late endosome/lysosome specific lipid, bis(monoacylglycero)phosphate (also called as ... More
An actin-associated protein present in the microtubule organizing center and the growth cones of PC-12 cells.
AuthorsBearer EL
JournalJ Neurosci
PubMed ID1372044
'The pathfinding ability of the growth cone depends upon the integrity of a dynamic actin filament network. However, although a number of actin-binding proteins have been found in growth cones, it is not known how these proteins come to be concentrated there or how they might interact to produce these ... More
Taurocholate induces pericanalicular localization of C6-NBD-ceramide in isolated hepatocyte couplets.
AuthorsCrawford JM, Vinter DW, Gollan JL
JournalAm J Physiol
PubMed ID1987800
'The mechanisms and pathways involved in hepatocellular transport of lipid destined for biliary excretion remain poorly understood. Using fluorescence microscopy of rat hepatocyte couplets in primary culture, we examined the effects of taurocholate (TC) on the intracellular distribution of 6-N-[7-nitrobenz-2-oxa-1,3-diazol- 4-yl]aminocaproyl-sphingosine (C6-NBD-ceramide), a lipid that accumulates in the Golgi apparatus. ... More
ADPKD: a human disease altering Golgi function and basolateral exocytosis in renal epithelia.
AuthorsCharron AJ, Bacallao RL, Wandinger-Ness A
JournalTraffic
PubMed ID11208155
'Epithelial cells explanted from autosomal dominant polycystic kidney disease (ADPKD) tissue exhibit impaired exocytosis, specifically between the Golgi and basolateral membrane (Charron A, Nakamura B, Bacallo R, Wandinger-Ness A. Compromised cytoarchitecture and polarized trafficking in autosomal dominant polycystic kidney disease cells. J Cell Biol 2000; 148: 111-124.). Here the defect ... More
Fluorescence-imaged microdeformation of the outer hair cell lateral wall.
AuthorsOghalai JS, Patel AA, Nakagawa T, Brownell WE
JournalJ Neurosci
PubMed ID9412485
'Outer hair cell (OHC) electromotility appears to be central to mammalian hearing and originates within its lateral wall. The OHC lateral wall is a unique trilaminate structure consisting of the plasma membrane (PM), the cortical lattice (CL), and the subsurface cisternae (SSC). We selectively labeled and imaged the lateral wall ... More
Ceramide as a second messenger: sticky solutions to sticky problems.
AuthorsVenkataraman K, Futerman AH
JournalTrends Cell Biol
PubMed ID10998592
'Much discussion has recently centred around the biochemical mechanisms by which ceramide is produced in signalling pathways. Since ceramide is virtually insoluble in aqueous solutions, the biological effects of ceramide should be considered in the context of its generation within the membrane lipid bilayer. To this end, we now summarize ... More
Reversed-phase liquid chromatographic method for the determination of 7-nitrobenz-2-oxa-1,3-diazol-4-yl-labelled lipid analogues.
AuthorsFrijters CM, Tuijn CJ, Hoek FJ, Groen AK, Oude Elferink RP, Zegers BN
JournalJ Chromatogr B Biomed Sci Appl
PubMed ID9686866
'This paper reports the development of a dual column system for the simultaneous separation of fluorescent short-chain ceramide, 6-[(7-nitrobenz-2-oxa-1,3,-diazol-4-yl[NBD])amino]hexanoyl-sphingo sine and its metabolites, C6-NBD-sphingomyelin and C6-NBD-glucosylceramide, as well as the fluorescent derivatives of choline and serine phosphatides. The method enables the separation of these lipids in a single run on ... More
Fluorescence assay of glucosylceramide glucosidase using NBD-cerebroside.
AuthorsAbe A, Shayman JA, Radin NS
JournalLipids
PubMed ID1487952
'A sensitive fluorometric assay for glucocerebroside beta-glucosidase [Dinur, T., Grabowski, G.A., Desnick, R.J., and Gatt, S. (1984) Anal. Biochem. 136, 223-234] has been reexamined. It was found that the lipids containing the NBD moiety (12-[N-methyl-N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)] used for standardization of the assay are light-sensitive and that the yield of fluorescent light ... More
Effects of mepanipyrim on intracellular trafficking: a comparative study on its effects on exocytic and endocytic trafficking of proteins, sphingolipids, and cholesterol.
AuthorsMiura I, Muroi M, Shiragami N, Yamaguchi I, Takatsuki A
JournalBiosci Biotechnol Biochem
PubMed ID8987670
'Mepanipyrim, N-(4-methyl-6-prop-1-ynylpyrimidin-2-yl)aniline, diminished the cell surface expression of envelope glycoproteins of Newcastle disease and vesicular stomatitis viruses at concentrations where their synthesis was not profoundly affected. Intoxication by diphtheria toxin and ricin and recycling of transferrin were not affected even when cells were treated with mepanipyrim for 2 h before ... More
Charge-based heterogeneity of human plasma lipoproteins at hypertriglyceridemia: capillary isotachophoresis study.
AuthorsDergunov AD, Hoy A, Smirnova EA, Visvikis S, Siest G
JournalInt J Biochem Cell Biol
PubMed ID12565713
'To reveal the metabolic links between and within pools of pro-atherogenic triglyceride(TG)-rich lipoproteins and anti-atherogenic high density lipoproteins (HDL), the changes in lipoprotein profile at hypertriglyceridemia were analyzed by capillary isotachophoresis. Plasma samples from patients with apoE3/3 phenotype were stained with a fluorescent probe NBD-C6-ceramide and lipoproteins resolved into six ... More
Uptake and metabolism of fluorescent lipid analogs by Pneumocystis carinii.
AuthorsSleight RG, Mehta MA, Kaneshiro ES
JournalJ Eukaryot Microbiol
PubMed ID7804204
Role of lysosomal acid ceramidase in the metabolism of ceramide in human skin fibroblasts.
AuthorsChen WW, Moser AB, Moser HW
JournalArch Biochem Biophys
PubMed ID7259198
A fluorescent derivative of ceramide: physical properties and use in studying the Golgi apparatus of animal cells.
AuthorsPagano RE
JournalMethods Cell Biol
PubMed ID2913469
The Golgi apparatus: insights from lipid biochemistry.
AuthorsPagano RE
JournalBiochem Soc Trans
PubMed ID2197129
Lipid sorting in epithelial cells.
AuthorsSimons K, van Meer G
JournalBiochemistry
PubMed ID3064805
Membrane glycolipid trafficking in living, polarized pancreatic acinar cells: assessment by confocal microscopy.
AuthorsCornell-Bell AH, Otake LR, Sadler K, Thomas PG, Lawrence S, Olsen K, Gumkowski F, Peterson JR, Jamieson JD
JournalMethods Cell Biol
PubMed ID8246783
Trafficking of glycosphingolipids in eukaryotic cells; sorting and recycling of lipids.
AuthorsHoekstra D, Kok JW
JournalBiochim Biophys Acta
PubMed ID1450202
Suicidal membrane repair regulates phosphatidylserine externalization during apoptosis.
One of the hallmarks of apoptosis is the redistribution of phosphatidylserine (PS) from the inner-to-outer plasma membrane (PM) leaflet, where it functions as a ligand for phagocyte recognition and the suppression of inflammatory responses. The mechanism by which apoptotic cells externalize PS has been assumed to involve ... More
Defining lipid transport pathways in animal cells.
AuthorsPagano RE, Sleight RG
JournalScience
PubMed ID4035344
A new technique for studying the metabolism and intracellular transport of lipid molecules in living cells based on the use of fluorescent lipid analogs is described. The cellular processing of various intermediates (phosphatidic acid and ceramide) and end products (phosphatidylcholine and phosphatidylethanolamine) in lipid biosynthesis is reviewed and a working ... More
To boldly glow ... applications of laser scanning confocal microscopy in developmental biology.
AuthorsPaddock SW
JournalBioessays
PubMed ID8024544
The laser scanning confocal microscope (LSCM) is now established as an invaluable tool in developmental biology for improved light microscope imaging of fluorescently labelled eggs, embryos and developing tissues. The universal application of the LSCM in biomedical research has stimulated improvements to the microscopes themselves and the synthesis of novel ... More
Activation of myosin V-based motility and F-actin-dependent network formation of endoplasmic reticulum during mitosis.
AuthorsWollert T, Weiss DG, Gerdes HH, Kuznetsov SA
JournalJ Cell Biol
PubMed ID12438410
It is widely believed that microtubule- and F-actin-based transport of cytoplasmic organelles and membrane fusion is down-regulated during mitosis. Here we show that during the transition of Xenopus egg extracts from interphase to metaphase myosin V-driven movement of small globular vesicles along F-actin is strongly inhibited. In contrast, the movement ... More
Sphingomyelin is synthesized at the plasma membrane of oligodendrocytes and by purified myelin membranes: a study with fluorescent- and radio-labelled ceramide analogues.
AuthorsVos JP, Giudici ML, van der Bijl P, Magni P, Marchesini S, van Golde LM, Lopes-Cardozo M
JournalFEBS Lett
PubMed ID7628646
In most cell types sphingomyelin is synthesized predominantly in the cis-medial compartments of the Golgi stacks whereas the contribution of the plasma membrane is much lower. The aim of this study was to assess the contribution of both compartments to the synthesis of sphingomyelin in myelinating cells. Therefore, oligodendrocytes from ... More
Inhibition of yeast inositol phosphorylceramide synthase by aureobasidin A measured by a fluorometric assay.
AuthorsZhong W, Murphy DJ, Georgopapadakou NH
JournalFEBS Lett
PubMed ID10606729
Inositol phosphorylceramide synthase (IPC synthase) is an essential and unique enzyme in fungal sphingolipid biosynthesis and is the target of the cyclic nonadepsipeptide antibiotic aureobasidin A. As a first step towards understanding the mechanism of aureobasidin A inhibition, we developed a fluorometric HPLC assay for IPC synthase using the Saccharomyces ... More
Possible involvement of heterotrimeric G proteins in the organization of the Golgi apparatus.
AuthorsYamaguchi T, Yamamoto A, Furuno A, Hatsuzawa K, Tani K, Himeno M, Tagaya M
JournalJ Biol Chem
PubMed ID9312142
Nordihydroguaiaretic acid (NDGA) caused disassembly of the Golgi apparatus of NRK cells in a dose-, time-, and energy-dependent manner but not in a microtubule-dependent manner. In contrast to brefeldin A, NDGA did not cause release of beta-COP, a component of Golgi-derived vesicles. However, NDGA-induced disassembly was blocked by AlF4-, an ... More
Assessment of golgi apparatus versus plasma membrane-localized multi-drug resistance-associated protein 1.
AuthorsKaufmann AM, Toro-Ramos AJ, Krise JP,
JournalMol Pharm
PubMed ID18557629
Traditionally, proteins belonging to the ATP-binding cassette superfamily have been thought to function exclusively at the plasma membrane (PM) of cells. We have previously shown multidrug resistance-associated protein 1 (MRP1) to reside on the Golgi apparatus of the multidrug resistant (MDR) human leukemic cell line HL-60 (HL-60/ADR); however, neither the ... More
Stable Golgi-mitochondria complexes and formation of Golgi Ca(2+) gradients in pancreatic acinar cells.
We have determined the localization of the Golgi with respect to other organelles in living pancreatic acinar cells and the importance of this localization to the establishment of Ca(2+) gradients over the Golgi. Using confocal microscopy and the Golgi-specific fluorescent probe 6-((N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino)hexanoyl)sphingosine, we found Golgi structures localizing to the outer ... More
Cholesterol controls lipid endocytosis through Rab11.
AuthorsTakahashi M, Murate M, Fukuda M, Sato SB, Ohta A, Kobayashi T,
JournalMol Biol Cell
PubMed ID17475773
Cellular cholesterol increases when cells reach confluency in Chinese hamster ovary (CHO) cells. We examined the endocytosis of several lipid probes in subconfluent and confluent CHO cells. In subconfluent cells, fluorescent lipid probes including poly(ethylene glycol)derivatized cholesterol, 22-(N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino)-23,24-bisnor-5-cholen-3beta-ol, and fluorescent sphingomyelin analogs were internalized to pericentriolar recycling endosomes. This accumulation ... More
ER-Tracker dye and BODIPY-brefeldin A differentiate the endoplasmic reticulum and golgi bodies from the tubular-vacuole system in living hyphae of Pisolithus tinctorius.
AuthorsCole L, Davies D, Hyde GJ, Ashford AE
JournalJ Microsc
PubMed ID10692127
Two fluorochromes, ER-TrackerTM Blue-White DPX dye and the fluorescent brefeldin A (BFA) derivative, BODIPY-BFA, label the endoplasmic reticulum (ER) in hyphal tips of Pisolithus tinctorius and allow its differentiation from the tubular-vacuole system at the light microscope level in living cells. The ER-Tracker dye labels a reticulate network similar in ... More
ATP-dependent fusion of liposomes with the Golgi apparatus of perforated cells.
AuthorsKobayashi T, Pagano RE
JournalCell
PubMed ID3191530
We examined the interactions of lipid vesicles (liposomes) labeled with various fluorescent markers with the intracellular membranes of semi-intact ("perforated") fibroblasts. When incubations were performed in the presence of an ATP-regenerating system, both vesicle lipids and entrapped water soluble markers were transferred to the Golgi apparatus of treated cells, indicative ... More
Microsurgical removal of centrosomes blocks cell reproduction and centriole generation in BSC-1 cells.
AuthorsManiotis A, Schliwa M
JournalCell
PubMed ID1934057
We have removed the centrosome from cultured BSC-1 cells by microsurgery, leaving enough cytoplasm with the nucleated cell fragment (karyoplast) to ensure survival and growth. In each experiment, we followed the fate of the karyoplast as well as the anucleate cell fragment (cytoplast) containing the original pair of centrioles. Experimental ... More
Fluorescent lipid probes: some properties and applications (a review).
AuthorsMaier O, Oberle V, Hoekstra D
JournalChem Phys Lipids
PubMed ID12093532
Odd as it may seem, experimental challenges in lipid research are often hampered by the simplicity of the lipid structure. Since, as in protein research, mutants or overexpression of lipids are not realistic, a considerable amount of lipid research relies on the use of tagged lipid analogues. However, given the ... More
Chemistry and biology of N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)-labeled lipids: fluorescent probes of biological and model membranes.
AuthorsChattopadhyay A
JournalChem Phys Lipids
PubMed ID2191793
Lipids that are covalently labeled with the 7-nitrobenz-2-oxa-1,3-diazol-4-yl (NBD) group are widely used as fluorescent analogues of native lipids in model and biological membranes to study a variety of processes. The fluorescent NBD group may be attached either to the polar or the apolar regions of a wide variety of ... More
Glucosylceramide synthesis is required for basic fibroblast growth factor and laminin to stimulate axonal growth.
AuthorsBoldin S, Futerman AH
JournalJ Neurochem
PubMed ID9003082
To test the hypothesis that neuronal growth requires the synthesis and supply of new membrane components to the growing neurite, we have examined the relationship between the synthesis of sphingolipids and the ability of two growth factors, basic fibroblast growth factor (bFGF) and laminin, to stimulate axonal growth in cultured ... More
Chimeric forms of furin and TGN38 are transported with the plasma membrane in the trans-Golgi network via distinct endosomal pathways.
AuthorsMallet WG, Maxfield FR
JournalJ Cell Biol
PubMed ID10465644
Furin and TGN38 are menbrane proteins that cycle between the plasma membrane and the trans-Golgi network (TGN), each maintaining a predominant distribution in the TGN. We have used chimeric proteins with an extracellular Tac domain and the cytoplasmic domain of TGN38 or furin to study the trafficking of these proteins ... More
Functional reconstitution of sphingomyelin synthase in Chinese hamster ovary cell membranes.
AuthorsHanada K, Horii M, Akamatsu Y
JournalBiochim Biophys Acta
PubMed ID1657181
Sphingomyelin synthase (phosphatidylcholine:ceramide phosphocholinetransferase) activity in the membranes of Chinese hamster ovary cells was found to be detectable with a fluorescent ceramide analog, containing a short acyl chain, as a substrate. We developed a method for the functional reconstitution of sphingomyelin synthase in detergent-treated membranes. Treatment of membranes with 1.5% ... More
In vivo metabolism of fluorescent ceramide in central nervous system myelin of adult rats.
AuthorsDi Biase A, Argiolas L, Confaloni A, Salvati S
JournalNeurochem Res
PubMed ID1754031
The metabolism of sphingolipids in the central nervous system (CNS) has been studied in adult rats by intraventricular administration of fluorescent ceramide (CER). Rats were sacrificed at various time points post inoculation and the fluorescence of CER, cerebrosides (CB), sulfatides (SULF) and sphingomyelin (SPM) was determined in the CNS myelin ... More