Authorsde Anda FC, Pollarolo G, Da Silva JS, Camoletto PG, Feiguin F, Dotti CG
JournalNature
PubMed ID16079847
'Neuronal polarization occurs shortly after mitosis. In neurons differentiating in vitro, axon formation follows the segregation of growth-promoting activities to only one of the multiple neurites that form after mitosis. It is unresolved whether such spatial restriction makes use of an intrinsic program, like during C. elegans embryo polarization, or ... More
AuthorsSnow CJ, Goody M, Kelly MW, Oster EC, Jones R, Khalil A, Henry CA,
JournalPLoS Genet
PubMed ID18833302
'Skeletal muscle morphogenesis transforms short muscle precursor cells into long, multinucleate myotubes that anchor to tendons via the myotendinous junction (MTJ). In vertebrates, a great deal is known about muscle specification as well as how somitic cells, as a cohort, generate the early myotome. However, the cellular mechanisms that generate ... More
Two-photon fluorescence absorption and emission spectra of dyes relevant for cell imaging.
AuthorsBestvater F, Spiess E, Stobrawa G, Hacker M, Feurer T, Porwol T, Berchner-Pfannschmidt U, Wotzlaw C, Acker H
JournalJ Microsc
PubMed ID12423261
'Two-photon absorption and emission spectra for fluorophores relevant in cell imaging were measured using a 45 fs Ti:sapphire laser, a continuously tuneable optical parametric amplifier for the excitation range 580-1150 nm and an optical multichannel analyser. The measurements included DNA stains, fluorescent dyes coupled to antibodies as well as organelle ... More
Use of fluorescent dyes for measurement and localization of organelles associated with Ca2+ store release in human neutrophils.
AuthorsDavies EV, Blanchfield H, Hallett MB
JournalCell Biol Int
PubMed ID9693835
'Fura-2 and its lipid analogue, FFP-18, were used to measure changes in cytosolic free Ca2+ concentration within human neutrophils. Whereas fura-2 was employed to monitor cytosolic Ca2+ increases throughout the cytosol, FFP-18 was used to monitor Ca2+ changes only near the membrane. This latter probe was incorporated into the plasma ... More
Changes in architecture of the Golgi complex and other subcellular organelles during myogenesis.
AuthorsRalston E
JournalJ Cell Biol
PubMed ID7678420
'Myogenesis involves changes in both gene expression and cellular architecture. Little is known of the organization, in muscle in vivo, of the subcellular organelles involved in protein synthesis despite the potential importance of targeted protein synthesis for formation and maintenance of functional domains such as the neuromuscular junction. A panel ... More
Porphyrin-retinamides: synthesis and cellular studies.
AuthorsSibrian-Vazquez M, Jensen TJ, Vicente MG
JournalBioconjug Chem
PubMed ID17518439
'A series of four porphyrin-retinamides containing either all-trans- or 13-cis-retinoid acid residues, directly linked to the para-phenyl position of meso-tetraphenylporphyrin or via a low-molecular-weight PEG spacer, have been synthesized. The biological properties of these conjugates were evaluated in a model cell line, human HEp2, and in neuroblastoma SK-N-DZ cells, which ... More
Cytoarchitecture of size-excluding compartments in living cells.
AuthorsProvance DW, McDowall A, Marko M, Luby-Phelps K
JournalJ Cell Sci
PubMed ID7980739
'By fluorescence ratio imaging of large and small inert tracer particles in living cells, we have previously shown that particles 24 nm in radius are excluded from otherwise uncharacterized compartments in the distal and perinuclear cytoplasm (Luby-Phelps, K. and Taylor, D.L., 1988. Cell Motil. Cytoskel. 10, 28-37). In this study ... More
Common trafficking pathway for variant antigens destined for the surface of the Plasmodium falciparum-infected erythrocyte.
AuthorsHaeggström M, Kironde F, Berzins K, Chen Q, Wahlgren M, Fernandez V
JournalMol Biochem Parasitol
PubMed ID14668007
'Intraerythrocytic Plasmodium falciparum exports proteins to the cytosol and to the plasma membrane of the host cell. We here present data revealing the existence of a unique common pathway for the surface bound traffic of the clonally variant antigens, repeated-interspersed-antigen (RIFINS) and P. falciparum erythrocyte-membrane-protein-1 (PfEMP1). RIFIN- and PfEMP1-specific antibodies ... More
Ceramide involvement in homocamptothecin- and camptothecin-induced cytotoxicity and apoptosis in colon HT29 cells.
AuthorsChauvier D, Morjani H, Manfait M
JournalInt J Oncol
PubMed ID11894136
'Topoisomerase I inhibitors of the camptothecin (CPT) family have emerged as potent clinical chemotherapeutic agents in first-line treatment of solid colorectal cancer and in second-line for 5-fluorouracil resistant patients. CPT and homocamptothecin (hCPT), derivative with enhanced lactone stability, induced growth inhibition in HT29 cells via p53-independent apoptosis. hCPT- and CPT-induced ... 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
Golgi tubule traffic and the effects of brefeldin A visualized in living cells.
AuthorsSciaky N, Presley J, Smith C, Zaal KJ, Cole N, Moreira JE, Terasaki M, Siggia E, Lippincott-Schwartz J
JournalJ Cell Biol
PubMed ID9382862
'The Golgi complex is a dynamic organelle engaged in both secretory and retrograde membrane traffic. Here, we use green fluorescent protein-Golgi protein chimeras to study Golgi morphology in vivo. In untreated cells, membrane tubules were a ubiquitous, prominent feature of the Golgi complex, serving both to interconnect adjacent Golgi elements ... 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
Measurement of spontaneous transfer and transbilayer movement of BODIPY-labeled lipids in lipid vesicles.
AuthorsBai J, Pagano RE
JournalBiochemistry
PubMed ID9220970
'An assay was developed to study the spontaneous transfer and transbilayer movement (flip-flop) of lipid analogs labeled with the fluorescent fatty acid, 5-(5,7-dimethyl BODIPY)-1-pentanoic acid (C5-DMB-) in large unilamellar lipid vesicles comprised of 1-palmitoyl-2-oleoyl phosphatidylcholine (POPC). The assay is based on the concentration-dependent changes in fluorescence intensity that occur when ... More
AuthorsSteinert S, Lee E, Tresset G, Zhang D, Hortsch R, Wetzel R, Hebbar S, Sundram JR, Kesavapany S, Boschke E, Kraut R,
JournalPLoS ONE
PubMed ID18716682
'BACKGROUND: The uptake and intracellular trafficking of sphingolipids, which self-associate into plasma membrane microdomains, is associated with many pathological conditions, including viral and toxin infection, lipid storage disease, and neurodegenerative disease. However, the means available to label the trafficking pathways of sphingolipids in live cells are extremely limited. In order ... More
Evidence for a cholesterol transport pathway from lysosomes to endoplasmic reticulum that is independent of the plasma membrane.
AuthorsUnderwood KW, Jacobs NL, Howley A, Liscum L
JournalJ Biol Chem
PubMed ID9461625
'We have studied the movement of low density lipoprotein (LDL)-derived cholesterol in cultured Chinese hamster ovary cells. Our hypothesis is that when LDL cholesterol is effluxed from lysosomes, the bulk of LDL cholesterol is mobilized to the plasma membrane, while another pathway delivers LDL cholesterol from lysosomes to acyl-CoA/cholesterol acyltransferase ... More
The mechanism of facilitated cell membrane resealing.
AuthorsTogo T, Alderton JM, Bi GQ, Steinhardt RA
JournalJ Cell Sci
PubMed ID9973606
'Disruption of the plasma membrane evokes an exocytotic response that is required for rapid membrane resealing. We show here in Swiss 3T3 fibroblasts that a second disruption at the same site reseals more rapidly than the initial wound. This facilitated response of resealing was inhibited by both low external Ca2+ ... More
Measurement of the rate of myelination using a fluorescent analogue of ceramide.
AuthorsBilderback TR, Chan JR, Harvey JJ, Glaser M
JournalJ Neurosci Res
PubMed ID9285525
'Fluorescence digital imaging microscopy was used to investigate the process of myelin formation by Schwann cells in neuronal cocultures. The uptake of the fluorescent ceramide analogue N-[5-(5,7-dimethyl BODIPY)-1-pentanoyl]D-erythro-sphingosine (C5-DMB-ceramide) and its return to the plasma membrane as the corresponding fluorescent sphingomyelin and galactocerebroside analogues were measured. Through observation of this ... More
TbRAB18, a developmentally regulated Golgi GTPase from Trypanosoma brucei.
AuthorsJeffries TR, Morgan GW, Field MC
JournalMol Biochem Parasitol
PubMed ID11985863
'The trypanosomal secretory system is broadly similar to that of higher eukaryotes as proteins enter the system via the endoplasmic reticulum and are transported to the Golgi complex for elaboration of glycan chains. Importantly N-glycan processing is stage specific with only the bloodstream form (BSF) processing beyond the oligomannose form. ... More
Role of KIFC3 motor protein in Golgi positioning and integration.
AuthorsXu Y, Takeda S, Nakata T, Noda Y, Tanaka Y, Hirokawa N
JournalJ Cell Biol
PubMed ID12135985
'KIFC3, a microtubule (MT) minus end-directed kinesin superfamily protein, is expressed abundantly and is associated with the Golgi apparatus in adrenocortical cells. We report here that disruption of the kifC3 gene induced fragmentation of the Golgi apparatus when cholesterol was depleted. Analysis of the reassembly process of the Golgi apparatus ... More
A novel fluorescent ceramide analogue for studying membrane traffic in animal cells: accumulation at the Golgi apparatus results in altered spectral properties of the sphingolipid precursor.
AuthorsPagano RE, Martin OC, Kang HC, Haugland RP
JournalJ Cell Biol
PubMed ID2045412
'A series of ceramide analogues bearing the fluorophore boron dipyrromethene difluoride (BODIPY) were synthesized and evaluated as vital stains for the Golgi apparatus, and as tools for studying lipid traffic between the Golgi apparatus and the plasma membrane of living cells. Studies of the spectral properties of several of 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
Plasmodium vivax and Plasmodium chabaudi: intraerythrocytic traffic of antigenically homologous proteins involves a brefeldin A-sensitive secretory pathway.
AuthorsBracho C, Dunia I, Romano M, Benedetti EL, Perez HA
JournalEur J Cell Biol
PubMed ID11302521
'We have used a monoclonal antibody (mAb 7C5B71) raised against the erythrocytic stages of Plasmodium vivax to identify a 148-kDa P vivax protein antigen (Pv-148) which crossreacts with an antigenically homologous 190-kDa protein of P. chabaudi (Pc-190). During parasite intraerythrocytic development Pv-148 and Pc-190 are exported into the host cell ... 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
Sphingolipid synthesis as a target for chemotherapy against malaria parasites.
AuthorsLauer SA, Ghori N, Haldar K
JournalProc Natl Acad Sci U S A
PubMed ID7568097
'The human malaria parasite Plasmodium falciparum contains sphingomyelin synthase in its Golgi apparatus and in a network of tubovesicular membranes in the cytoplasm of the infected erythrocyte. Palmitoyl and decanoyl analogues of 1-phenyl-2-acylamino-3-morpholino-1-propanol inhibit the enzyme activity in infected erythrocytes. An average of 35% of the activity is extremely sensitive ... More
Fluorescence and multiphoton imaging resolve unique structural forms of sterol in membranes of living cells.
'Although cholesterol is an essential component of mammalian membranes, resolution of cholesterol organization in membranes and organelles (i.e. lysosomes) of living cells is hampered by the paucity of nondestructive, nonperturbing methods providing real time structural information. Advantage was taken of the fact that the emission maxima of a naturally occurring ... More
Modified low density lipoprotein delivers substrate for ceramide formation and stimulates the sphingomyelin-ceramide pathway in human macrophages.
AuthorsKinscherf R, Claus R, Deigner HP, Nauen O, Gehrke C, Hermetter A, Russwurm S, Daniel V, Hack V, Metz J
JournalFEBS Lett
PubMed ID9094423
'Exposure of human blood monocytes derived macrophages to modified (oxidized or acetylated) LDL induced a approximately 40% elevation (60 pmol/10(6) cells) of the endogenous level of the sphingolipid ceramide. A rise of both neutral and acidic SMase activity was found after treatment with oxidized LDL (250 and 80%), while addition ... 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
Applications of ratio fluorescence microscopy in the study of cell physiology.
AuthorsDunn KW, Mayor S, Myers JN, Maxfield FR
JournalFASEB J
PubMed ID8005385
'Quantitative fluorescence microscopy is becoming an increasingly important tool in the study of cell biology. Fluorescence microscopy has long been used for qualitative characterizations of subcellular distributions of proteins, lipids, nucleic acids, and ions, but quantifying these distributions is complicated by a variety of optical, biological, and physical factors. Many ... More
Fluorescent probes for living cells.
AuthorsJohnson I
JournalHistochem J
PubMed ID10188922
'The functional characteristics of fluorescent probes used for imaging and measuring dynamic processes in living cells are reviewed. Initial consideration is given to general design requirements for delivery, targeting, detectability and fluorescence readout, and current technologies for attaining them. Discussion then proceeds to the more application-specific properties of intracellular ion ... More
Septin-dependent compartmentalization of the endoplasmic reticulum during yeast polarized growth.
AuthorsLuedeke C, Frei SB, Sbalzarini I, Schwarz H, Spang A, Barral Y
JournalJ Cell Biol
PubMed ID15967812
'Polarized cells frequently use diffusion barriers to separate plasma membrane domains. It is unknown whether diffusion barriers also compartmentalize intracellular organelles. We used photobleaching techniques to characterize protein diffusion in the yeast endoplasmic reticulum (ER). Although a soluble protein diffused rapidly throughout the ER lumen, diffusion of ER membrane proteins ... More
Golgi apparatus dynamics during mouse oocyte in vitro maturation: effect of the membrane trafficking inhibitor brefeldin A.
AuthorsMoreno RD, Schatten G, Ramalho-Santos J
JournalBiol Reprod
PubMed ID11967185
'We have studied Golgi apparatus dynamics during mouse oocyte in vitro maturation, employing both live imaging with the fluorescent lipid BODIPY-ceramide and immunocytochemistry using several specific markers (beta-COP, giantin, and TGN38). In germinal vesicle oocytes the Golgi consisted of a series of structures, possibly cisternal stacks, dispersed in the ooplasm, ... More
Evidence for secretory pathway localization of a voltage-dependent anion channel isoform.
AuthorsBuettner R, Papoutsoglou G, Scemes E, Spray DC, Dermietzel R
JournalProc Natl Acad Sci U S A
PubMed ID10716730
'Voltage-dependent anion channels (VDACs) are pore-forming proteins (porins) that form the major pathway for movement of adenine nucleotides through the outer mitochondrial membrane. Electrophysiological studies indicate that VDAC-like channel activity is also prevalent in the cell membranes of many mammalian cells. However, the multitopological localization of porins outside the mitochondrion ... More
HVEM tomography of the trans-Golgi network: structural insights and identification of a lace-like vesicle coat.
AuthorsLadinsky MS, Kremer JR, Furcinitti PS, McIntosh JR, Howell KE
JournalJ Cell Biol
PubMed ID7929568
'High voltage electron microscopy and computer axial tomography have been used to study the 3-D structure of trans-Golgi cisternae and trans-Golgi networks (TGNs) in NRK cells. Both structures were specifically labeled by photoconversion of a fluorescent analogue of ceramide using a modification of the techique of Pagano et al. (J. ... More
ISC1-encoded inositol phosphosphingolipid phospholipase C is involved in Na+/Li+ halotolerance of Saccharomyces cerevisiae.
AuthorsBetz C, Zajonc D, Moll M, Schweizer E
JournalEur J Biochem
PubMed ID12180980
'In Saccharomyces cerevisiae, toxic concentrations of Na+ orLi+ ions induce the expression of the cation-extrusion ATPase gene, ENA1. Several well-studied signal transduction pathways are known correlating high salinity to the transcriptional activation of ENA1. Nevertheless, information on the actual sensing mechanism initiating these pathways is limited. Here, we report that ... More
Sphingomyelinase activity associated with human plasma low density lipoprotein.
'Isolated human plasma low density lipoprotein (LDL) was observed to possess sphingomyelinase activity. Accordingly, the formation of ceramide was catalyzed by LDL at 37 degrees C using tertiary liposomes composed of sphingomyelin (mole fraction (x) = 0.2), 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (x = 0.7), 1, 2-dimyristoyl-sn-glycero-3-phospho-rac-glycerol (x = 0.1), and either the fluorescent ... More
Lateral diffusion of small compounds in human stratum corneum and model lipid bilayer systems.
AuthorsJohnson ME, Berk DA, Blankschtein D, Golan DE, Jain RK, Langer RS
JournalBiophys J
PubMed ID8913603
'An image-based technique of fluorescence recovery after photobleaching (video-FRAP) was used to measure the lateral diffusion coefficients of a series of nine fluorescent probes in two model lipid bilayer systems, dimyristoylphosphatidylcholine (DMPC) and DMPC/cholesterol (40 mol%), as well as in human stratum corneum-extracted lipids. The probes were all lipophilic, varied ... More
The Golgi apparatus and the centrosome are localized to the sites of newly emerging axons in cerebellar granule neurons in vitro.
AuthorsZmuda JF, Rivas RJ
JournalCell Motil Cytoskeleton
PubMed ID9744296
'Cultured cerebellar granule neurons develop their characteristic axonal and dendritic morphologies in a series of discrete temporal steps highly similar to those observed in situ, initially extending a single process, followed by the extension of a second process from the opposite pole of the cell, both of which develop into ... More
Intracellular localization of the radiation enhancer motexafin gadolinium using interferometric Fourier fluorescence microscopy.
AuthorsWoodburn KW
JournalJ Pharmacol Exp Ther
PubMed ID11356908
'Motexafin gadolinium (MGd) is a unique therapeutic agent that localizes in cancer cells and increases tumor response to ionizing radiation and certain chemotherapeutics. The in vitro intracellular localization, accumulation, and retention of MGd in murine EMT6 mammary sarcoma and Rif-1 fibrosarcoma cell lines were studied using interferometric Fourier fluorescence microscopy. ... More
Flippase activity detected with unlabeled lipids by shape changes of giant unilamellar vesicles.
AuthorsPapadopulos A, Vehring S, López-Montero I, Kutschenko L, Stöckl M, Devaux PF, Kozlov M, Pomorski T, Herrmann A
JournalJ Biol Chem
PubMed ID17369612
'Transbilayer movement of phospholipids in biological membranes is mediated by energy-dependent and energy-independent flippases. Available methods for detection of flippase mediated transversal flip-flop are essentially based on spin-labeled or fluorescent lipid analogues. Here we demonstrate that shape change of giant unilamellar vesicles (GUVs) can be used as a new tool ... 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
Use of BODIPY-labeled sphingolipids to study membrane traffic along the endocytic pathway.
AuthorsPagano RE, Chen CS
JournalAnn N Y Acad Sci
PubMed ID9668349
'In this chapter we discuss the use of BODIPY-labeled sphingolipids to study lipid transport along the endocytic pathway of cultured mammalian cells. The unique spectral properties of the BODIPY fluorophore allow the investigator to distinguish various populations of labeled endosome and lysosomes within the living cell by fluorescence microscopy, and ... More
A membrane network for nutrient import in red cells infected with the malaria parasite.
AuthorsLauer SA, Rathod PK, Ghori N, Haldar K
JournalScience
PubMed ID9148808
'The human malaria parasite Plasmodium falciparum exports an interconnected network of tubovesicular membranes (the TVM) that extends from the parasite''s vacuolar membrane to the periphery of the red cell. Here it is shown that extracellular solutes such as Lucifer yellow enter the TVM and are delivered to the parasite. Blocking ... More
Distribution and biophysical properties of fluorescent lipids on the surface of adult Schistosoma mansoni.
AuthorsRedman CA, Kusel JR
JournalParasitology
PubMed ID8786886
'The properties of 4 fluorescent lipid compounds in the surface membrane of adult male Schistosoma mansoni worms were examined by fluorescent microscopy and fluorescent recovery after photobleaching (FRAP). The data suggest that the probes N-(4,4-difluoro-5,7-dimethyl-4-bora-3a,4a-diaza-s- indacene-3-pentanoyl) sphingosine (BODIPY FL ceramide) and PKH2 pass through the outer membrane and enter structures ... More
Complete vesiculation of Golgi membranes and inhibition of protein transport by a novel sea sponge metabolite, ilimaquinone.
AuthorsTakizawa PA, Yucel JK, Veit B, Faulkner DJ, Deerinck T, Soto G, Ellisman M, Malhotra V
JournalCell
PubMed ID8513494
'We have identified a novel natural metabolite, ilimaquinone (IQ), from sea sponges that causes Golgi membranes to break down completely in vivo into small vesicular structures (called vesiculated Golgi membranes [VGMs]). Under these conditions, transport of newly synthesized proteins from endoplasmic reticulum (ER) to the cis-Golgi-derived VGMs is unaffected; however, ... More
Inheriting the Golgi.
AuthorsRoth MG
JournalCell
PubMed ID10612391
Analyzing morphogenetic cell behaviors in vitally stained zebrafish embryos.
AuthorsCooper MS, D'Amico LA, Henry CA
JournalMethods Mol Biol
PubMed ID10231793
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
Applications of BODIPY-sphingolipid analogs to study lipid traffic and metabolism in cells.
AuthorsPagano RE, Watanabe R, Wheatley C, Dominguez M
JournalMethods Enzymol
PubMed ID11070900
Use of Bodipy-labeled sphingolipid and cholesterol analogs to examine membrane microdomains in cells.
AuthorsMarks DL, Bittman R, Pagano RE,
JournalHistochem Cell Biol
PubMed ID18820942
Much evidence has accumulated to show that cellular membranes such as the plasma membrane, contain multiple
Essential role of MD-2 in LPS responsiveness and TLR4 distribution.
AuthorsNagai Y, Akashi S, Nagafuku M, Ogata M, Iwakura Y, Akira S, Kitamura T, Kosugi A, Kimoto M, Miyake K
JournalNat Immunol
PubMed ID12055629
Toll-like receptor 4 (TLR4) mediates lipopolysaccharide (LPS) signaling in a variety of cell types. MD-2 is associated with the extracellular domain of TLR4 and augments TLR4-dependent LPS responses in vitro. We show here that MD-2(-/-) mice do not respond to LPS, do survive endotoxic shock but are susceptible to Salmonella ... More
Cholesterol-independent targeting of Golgi membrane proteins in insect cells.
AuthorsRolls MM, Marquardt MT, Kielian M, Machamer CE
JournalMol Biol Cell
PubMed ID9362056
Distinct lipid compositions of intracellular organelles could provide a physical basis for targeting of membrane proteins, particularly where transmembrane domains have been shown to play a role. We tested the possibility that cholesterol is required for targeting of membrane proteins to the Golgi complex. We used insect cells for our ... More
Fluorescent labelling of intracellular bacteria in living host cells.
AuthorsBoleti H, Ojcius DM, Dautry-Varsat A
JournalJ Microbiol Methods
PubMed ID10802143
The fluorescent reagent, CellTracker, labels metabolically-active cells and was used here to label Chlamydia in vivo during their exponential phase of growth in infected cells. HeLa cells infected with C. psittaci were labelled with the CellTracker reagents between 15 and 48 h post-infection. The fluorescent label accumulated in the host-cell ... More
Carotenoids located in human lymphocyte subpopulations and natural killer cells by Raman microspectroscopy.
The presence and subcellular location of carotenoids in human lymphocyte subpopulations (CD4+, CD8+, T-cell receptor-gamma delta+, and CD19+) and natural killer cells (CD16+) were studied by means of Raman microspectroscopy. In CD4+ lymphocytes a high concentration (approximately 10(-3) M) of carotenoids was found in the Gall body. In CD8+ lymphocytes, ... More
Use of fluorescent sphingolipid analogs to study lipid transport along the endocytic pathway.
AuthorsMarks DL, Singh RD, Choudhury A, Wheatley CL, Pagano RE
JournalMethods
PubMed ID15905102
Sphingolipids (SLs) are concentrated at the plasma membrane where they play important roles in cell-cell communication, host-pathogen interactions, and cell signaling events. Our laboratory has used fluorescent SL analogs and SL-binding toxins to elucidate mechanisms by which SLs are internalized by endocytosis and subsequently sorted and transported to various intracellular ... More
In vivo time-lapse imaging of cell divisions during neurogenesis in the developing zebrafish retina.
AuthorsDas T, Payer B, Cayouette M, Harris WA
JournalNeuron
PubMed ID12597858
Two-photon excitation microscopy was used to reconstruct cell divisions in living zebrafish embryonic retinas. Contrary to proposed models for vertebrate asymmetric divisions, no apico-basal cell divisions take place in the zebrafish retina during the generation of postmitotic neurons. However, a surprising shift in the orientation of cell division from central-peripheral ... More
Cholesterol level regulates endosome motility via Rab proteins.
AuthorsChen H, Yang J, Low PS, Cheng JX,
JournalBiophys J
PubMed ID17981910
The role of cholesterol in the regulation of endosome motility was investigated by monitoring the intracellular trafficking of endocytosed folate receptors (FRs) labeled with fluorescent folate conjugates. Real-time fluorescence imaging of HeLa cells transfected with green fluorescent protein-tubulin revealed that FR-containing endosomes migrate along microtubules. Moreover, microinjection with antibodies that ... More
Glycosphingolipid synthesis requires FAPP2 transfer of glucosylceramide.
AuthorsD'Angelo G, Polishchuk E, Di Tullio G, Santoro M, Di Campli A, Godi A, West G, Bielawski J, Chuang CC, van der Spoel AC, Platt FM, Hannun YA, Polishchuk R, Mattjus P, De Matteis MA,
JournalNature
PubMed ID17687330
The molecular machinery responsible for the generation of transport carriers moving from the Golgi complex to the plasma membrane relies on a tight interplay between proteins and lipids. Among the lipid-binding proteins of this machinery, we previously identified the four-phosphate adaptor protein FAPP2, the pleckstrin homology domain of which binds ... More
Novel anti-cholesterol monoclonal immunoglobulin G antibodies as probes and potential modulators of membrane raft-dependent immune functions.
AuthorsBíró A, Cervenak L, Balogh A, Lorincz A, Uray K, Horváth A, Romics L, Matkó J, Füst G, László G,
JournalJ Lipid Res
PubMed ID17023738
Natural autoantibodies against cholesterol are present in the sera of all healthy individuals; their function, production, and regulation, however, are still unclear. Here, we managed to produce two monoclonal anti-cholesterol antibodies (ACHAs) by immunizing mice with cholesterol-rich liposomes. The new ACHAs were specific to cholesterol and to some structurally closely ... More
Uptake kinetics and intracellular localization of hypocrellin photosensitizers for photodynamic therapy: a confocal microscopy study.
AuthorsMiller GG, Brown K, Moore RB, Diwu ZJ, Liu J, Huang L, Lown JW, Begg DA, Chlumecky V, Tulip J
JournalPhotochem Photobiol
PubMed ID7568409
Hypocrellins are naturally occurring compounds with photosensitizing properties in biological systems. We have prepared synthetic derivatives of hypocrellin B, which have promise as photosensitizers in the clinical application of photodynamic therapy. The intracellular localization and uptake kinetics of hypocrellin B and several selected hypocrellin congeners were determined semiquantitatively by fluorescence ... More
A FRET-based fluorogenic phosphine for live-cell imaging with the Staudinger ligation.
AuthorsHangauer MJ, Bertozzi CR,
JournalAngew Chem Int Ed Engl
PubMed ID18306205
Fluorescent phosphine probes have been used for direct imaging of various azide-bearing biomolecules with the Staudinger ligation in cell-free environments. Recently, we applied phosphine-based dyes to image azides on the surface of live cells. Notably, significant labeling above background could only be achieved using a highly negatively charged ... More
A Plasmodium falciparum protein located in Maurer's clefts underneath knobs and protein localization in association with Rhop-3 and SERA in the intracellular network of infected erythrocytes.
AuthorsSam-Yellowe TY, Fujioka H, Aikawa M, Hall T, Drazba JA
JournalParasitol Res
PubMed ID11293564
We report on the characterization of monoclonal antibodies against Plasmodium falciparum schizonts, which recognize parasite proteins of 130 kDa and 20 kDa. The 130-kDa protein was released by alkaline sodium carbonate treatment, suggesting that the protein is a peripheral membrane protein, while the 20-kDa protein remained associated with the membranes ... More
Bioinformatic and image analyses of the cellular localization of the apoptotic proteins endonuclease G, AIF, and AMID during apoptosis in human cells.
AuthorsVarecha M, Amrichová J, Zimmermann M, Ulman V, Lukásová E, Kozubek M
JournalApoptosis
PubMed ID17347867
We studied the cellular localization of the apoptotic proteins endonuclease G, AIF, and AMID in silico using three prediction tools and in living cells using both single-cell colocalization image analysis and nuclear translocation analysis. We confirmed the mitochondrial localization of endonuclease G and AIF by prediction analysis and by single-cell ... More
Golgi membranes are absorbed into and reemerge from the ER during mitosis.
AuthorsZaal KJ, Smith CL, Polishchuk RS, Altan N, Cole NB, Ellenberg J, Hirschberg K, Presley JF, Roberts TH, Siggia E, Phair RD, Lippincott-Schwartz J
JournalCell
PubMed ID10612395
Quantitative imaging and photobleaching were used to measure ER/Golgi recycling of GFP-tagged Golgi proteins in interphase cells and to monitor the dissolution and reformation of the Golgi during mitosis. In interphase, recycling occurred every 1.5 hr, and blocking ER egress trapped cycling Golgi enzymes in the ER with loss of ... More
Tamoxifen inhibits acidification in cells independent of the estrogen receptor.
AuthorsAltan N, Chen Y, Schindler M, Simon SM
JournalProc Natl Acad Sci U S A
PubMed ID10200279
Tamoxifen has been reported to have numerous physiological effects that are independent of the estrogen receptor, including sensitization of resistant tumor cells to many chemotherapeutic agents. Drug-resistant cells sequester weak base chemotherapeutics in acidic organelles away from their sites of action in the cytosol and nucleus. This work reports that ... 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
Mechanism of cellular 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) reduction.
AuthorsLiu Y, Peterson DA, Kimura H, Schubert D
JournalJ Neurochem
PubMed ID9231715
3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) reduction is one of the most frequently used methods for measuring cell proliferation and neural cytotoxicity. It is widely assumed that MTT is reduced by active mitochondria in living cells. By using isolated mitochondria from rat brain and B12 cells, we indeed found that malate, glutamate, and ... More
Intracardiac fluid forces are an essential epigenetic factor for embryonic cardiogenesis.
AuthorsHove JR, Köster RW, Forouhar AS, Acevedo-Bolton G, Fraser SE, Gharib M
JournalNature
PubMed ID12520305
The pattern of blood flow in the developing heart has long been proposed to play a significant role in cardiac morphogenesis. In response to flow-induced forces, cultured cardiac endothelial cells rearrange their cytoskeletal structure and change their gene expression profiles. To link such in vitro data to the intact heart, ... More
Role of dynactin in endocytic traffic: effects of dynamitin overexpression and colocalization with CLIP-170.
AuthorsValetti C, Wetzel DM, Schrader M, Hasbani MJ, Gill SR, Kreis TE, Schroer TA
JournalMol Biol Cell
PubMed ID10588646
The flow of material from peripheral, early endosomes to late endosomes requires microtubules and is thought to be facilitated by the minus end-directed motor cytoplasmic dynein and its activator dynactin. The microtubule-binding protein CLIP-170 may also play a role by providing an early link to endosomes. Here, we show that ... More
Internalized plasma membrane cholesterol passes through an endosome compartment that is distinct from the acid vesicle-lysosome compartment.
AuthorsPorpaczy Z, Tomasek JJ, Freeman DA
JournalExp Cell Res
PubMed ID9260888
Cholesterol from the plasma membrane of MA-10 Leydig tumor cells is internalized into the cell and either esterified or used as substrate for steroid hormone synthesis. In the present studies we show that chloroquine and sphinganine cause LDL cholesterol and cholesteryl esters to accumulate in the cells. A lysosome fraction ... More
A proline-rich region and nearby cysteine residues target XLalphas to the Golgi complex region.
AuthorsUgur O, Jones TL
JournalMol Biol Cell
PubMed ID10749939
XLalphas is a splice variant of the heterotrimeric G protein, Galpha(s), found on Golgi membranes in cells with regulated and constitutive secretion. We examined the role of the alternatively spliced amino terminus of XLalphas for Golgi targeting with the use of subcellular fractionation and fluorescence microscopy. XLalphas incorporated [(3)H]palmitate, and ... More
Endoplasmic reticulum and Golgi apparatus are the preferential sites of Foscan localisation in cultured tumour cells.
AuthorsTeiten MH, Bezdetnaya L, Morlière P, Santus R, Guillemin F
JournalBr J Cancer
PubMed ID12556974
Intracellular photosensitiser localisation significantly influences the mechanism of response to photodynamic therapy (PDT), since the primary sites of damage are closely related to the specific sensitiser distribution. Foscan subcellular localisation in the MCF-7 human adenocarcinoma cell line has been studied by means of confocal microscopy and microspectrofluorometry. The fluorescence topographic ... More
Rapid, endoplasmic reticulum-independent diffusion of the mitotic Golgi haze.
AuthorsAxelsson MA, Warren G
JournalMol Biol Cell
PubMed ID14767069
Early in mitosis, the mammalian Golgi apparatus disassembles, and fluorescence microscopy reveals Golgi clusters and an extensive, nonresolvable haze that either represents scattered vesicles or a merged endoplasmic reticulum (ER)-Golgi compartment. To help decide between these alternatives, we have carried out a combined microscopic and pharmacological analysis, by using a ... More
A requirement for ARF6 in Fcgamma receptor-mediated phagocytosis in macrophages.
AuthorsZhang Q, Cox D, Tseng CC, Donaldson JG, Greenberg S
JournalJ Biol Chem
PubMed ID9685333
Phagocytosis requires extension of F-actin-rich pseudopods and is accompanied by membrane fusion events. Members of the ARF family of GTPases are essential for many aspects of membrane trafficking. To test a role for this family of proteins in Fcgamma receptor-mediated phagocytosis, we utilized the fungal metabolite brefeldin A (BFA). The ... More
C11-BODIPY(581/591), an oxidation-sensitive fluorescent lipid peroxidation probe: (micro)spectroscopic characterization and validation of methodology.
AuthorsDrummen GP, van Liebergen LC, Op den Kamp JA, Post JA
JournalFree Radic Biol Med
PubMed ID12160930
C11-BODIPY(581/591) is a fluorescent radio-probe for indexing lipid peroxidation and antioxidant efficacy in model membrane systems and living cells, with excellent characteristics: (i) emission in the visible range of the electromagnetic spectrum, with good spectral separation of the nonoxidized (595 nm) and oxidized (520 nm) forms; (ii) has a high ... More
Uptake and rapid transfer of fluorescent ceramide analogues to acidosomes (late endosomes) in Paramecium.
AuthorsIwamoto M, Allen RD
JournalJ Histochem Cytochem
PubMed ID15100234
The ciliated protozoan Paramecium incorporates sphingolipids into its cell membranes. However, it is still unclear if these sphingolipids are metabolically synthesized in the cell or if their precursors are taken up from exogenous materials. Here we studied the route of uptake of fluorescence-labeled analogues of ceramide. Fluorescent ceramide was taken ... More
A deficiency of the small GTPase rab8 inhibits membrane traffic in developing neurons.
AuthorsHuber LA, Dupree P, Dotti CG
JournalMol Cell Biol
PubMed ID7823956
One of the major activities of developing neurons is the transport of new membrane to the growing axon. Candidates for playing a key role in the regulation of this intense traffic are the small GTP-binding proteins of the rab family. We have used hippocampal neurons in culture and analyzed membrane ... More
Glycosylphosphatidylinositol biosynthetic enzymes are localized to a stable tubular subcompartment of the endoplasmic reticulum in Leishmania mexicana.
AuthorsIlgoutz SC, Mullin KA, Southwell BR, McConville MJ
JournalEMBO J
PubMed ID10393180
Glycosylphosphatidylinositols (GPI) are essential components in the plasma membrane of the protozoan parasite Leishmania mexicana, both as membrane anchors for the major surface macromolecules and as the sole class of free glycolipids. We provide evidence that L.mexicana dolichol-phosphate-mannose synthase (DPMS), a key enzyme in GPI biosynthesis, is localized to a ... More
Interconversion of sphingomyelin and ceramide in adult Schistosoma mansoni.
AuthorsRedman CA, Kennington S, Spathopoulou T, Kusel JR
JournalMol Biochem Parasitol
PubMed ID9497039
Fluorescent lipid analogues of the lipids ceramide and sphingomyelin, namely BODIPY FL C5-ceramide ((N-(4,4-difluoro-5,7-dimethyl-4-bora-3a,4a-diaza-sindacene-3-pentanoyl) sphingosine) and BODIPY C5-sphingomyelin ((N-(4,4-difluoro-5,7-dimethyl-4-bora-3a,4a-diaza-s-indacene-3-pentanoyl) sphingosyl phosphocholine), respectively, were used to investigate the presence of a sphingomyelin cycle in Schistosoma mansoni adult males. The parasites were able to convert BODIPY FL C5-ceramide into a fluorescent sphingomyelin ... More
HDL-mediated cholesterol uptake and targeting to lipid droplets in adipocytes.
AuthorsDagher G, Donne N, Klein C, Ferre P, Dugail I
JournalJ Lipid Res
PubMed ID12867544
Adipocytes express high levels of the HDL scavenger receptor class B type I in a differentiation-dependent manner. We thus have analyzed the routes of HDL cholesterol trafficking at different phases of adipocyte differentiation in the 3T3-L1 cell line. One novel and salient feature of this paper is the observation of ... More
"Synchronized" endocytosis and intracellular sorting in alveolar macrophages: the early sorting endosome is a transient organelle.
AuthorsWard DM, Perou CM, Lloyd M, Kaplan J
JournalJ Cell Biol
PubMed ID7775570
Incubation of alveolar macrophages in hypoosmotic K(+)-containing buffers results in persistent cell swelling and an inability to undergo regulatory volume decrease. We demonstrate that cells incubated in hypo-K+ show an inhibition of endocytosis without any observed alteration in recycling. The inhibition of endocytosis affected all forms of membrane internalization, receptor ... More
Cationic amphiphilic drugs inhibit the internalization of cholera toxin to the Golgi apparatus and the subsequent elevation of cyclic AMP.
AuthorsSofer A, Futerman AH
JournalJ Biol Chem
PubMed ID7744860
Cholera toxin (CT) consists of a pentameric B subunit which binds with high affinity to ganglioside GM1, and an A subunit which stimulates adenylate cyclase, resulting in the elevation of cAMP. We now examine the effect of cationic amphiphilic drugs (CADs) on the internalization of rhodamine (Rh)-CT in cultured hippocampal ... More
Separation of phospholipids in microfluidic chip device: application to high-throughput screening assays for lipid-modifying enzymes.
AuthorsLin S, Fischl AS, Bi X, Parce W
JournalAnal Biochem
PubMed ID12633607
Phospholipid molecules such as ceramide and phosphoinositides play crucial roles in signal transduction pathways. Lipid-modifying enzymes including sphingomyelinase and phosphoinositide kinases regulate the generation and degradation of these lipid-signaling molecules and are important therapeutic targets in drug discovery. We now report a sensitive and convenient method to separate these lipids ... More
Uptake and cellular localization of exogenous lipids by Giardia lamblia, a primitive eukaryote.
AuthorsStevens TL, Gibson GR, Adam R, Maier J, Allison-Ennis M, Das S
JournalExp Parasitol
PubMed ID9207743
Giardia lamblia trophozoites are unable to carry out de novo lipid synthesis. It is therefore likely that lipids are acquired from the small intestine of the host, in which the trophozoites are exposed to free and conjugated fatty acids, various sterols, phospholipids, bile acids, and bile-lipid mixed micelles. Here we ... More
Isolation, cloning, and characterization of a new mammalian coronin family member, coroninse, which is regulated within the protein kinase C signaling pathway.
In order to understand the regulatory role of protein kinase C (PKC) in secretory epithelia, it is necessary to identify and characterize specific downstream targets. We previously identified one such protein in studies of gastric parietal cells. This protein was referred to as pp66 because it migrated with an apparent ... More
Point mutation of adenosine triphosphate-binding motif generated rigor kinesin that selectively blocks anterograde lysosome membrane transport.
AuthorsNakata T, Hirokawa N
JournalJ Cell Biol
PubMed ID7490281
In the study of motor proteins, the molecular mechanism of mechanochemical coupling, as well as the cellular role of these proteins, is an important issue. To assess these questions we introduced cDNA of wild-type and site-directed mutant kinesin heavy chains into fibroblasts, and analyzed the behavior of the recombinant proteins ... More
Purification and characterization of UDP-glucose:ceramide glucosyltransferase from rat liver Golgi membranes.
AuthorsPaul P, Kamisaka Y, Marks DL, Pagano RE
JournalJ Biol Chem
PubMed ID8567691
We present a method for solubilizing and purifying UDP-Glc:ceramide glucosyltransferase (EC 2.4.1.80; glucosylceramide synthase (GCS) from a rat liver and present data on its substrate specificity. A Golgi membrane fraction was isolated, washed with N-lauroylsarcosine, and subsequently treated with 3[3-cholamidopropyl)-dimethylammonio]-2-hydroxy-1-propanesulfonate to solubilize the enzyme. GCS activity was monitored throughout purification ... More
Elimination of plasma membrane phosphatidylinositol (4,5)-bisphosphate is required for exocytosis from mast cells.
AuthorsHammond GR, Dove SK, Nicol A, Pinxteren JA, Zicha D, Schiavo G
JournalJ Cell Sci
PubMed ID16687737
The inositol lipid phosphatidylinositol (4,5)-bisphosphate [PtdIns(4,5)P2] is involved in a myriad of cellular processes, including the regulation of exocytosis and endocytosis. In this paper, we address the role of PtdIns(4,5)P2 in compound exocytosis from rat peritoneal mast cells. This process involves granule-plasma membrane fusion as well as homotypic granule membrane ... More
Brefeldin A and mannose 6-phosphate regulation of acrosomic related vesicular trafficking.
AuthorsWest AP, Willison KR
JournalEur J Cell Biol
PubMed ID8864659
Acrosomal biogenesis represents a unique system for the molecular analysis of the various processes involved in vesicular membrane transport. To assess the basic membrane trafficking mechanisms used in spermatids, we have used two fluorescent lipid compounds that label: a) the Golgi and Golgi-derived vesicles (C5-DMB-Cer), and b) endocytic vesicles (FM4-64). ... More
Astrocyte beta1-adrenergic receptor immunoreactivity and agonist induced increases in [Ca2+]i: differential results indicative of a modified membrane receptor.
AuthorsThorlin T, Anders P, Persson I, Eriksson PS, Rönnbäck L, Hansson E
JournalLife Sci
PubMed ID10972197
Antibodies against the C-terminus of the beta1-adrenergic receptor were used for staining cultured astrocytes from the rat cerebral cortex. Immunoreactivity was found to be localized exclusively to an intracellular organelle structure similar to the Golgi complex, with no staining of the plasma membrane. The astrocytes stained positive with BODIPY CGP ... More
Ceramide in nitric oxide inhibition of glioma cell growth. Evidence for the involvement of ceramide traffic.
The treatment of C6 glioma cells with the nitric oxide donor, PAPANONOate ((Z)-[N-(3-ammoniopropyl)-N-(n-propyl)amino]diazen-1-ium-1,2-diolate), resulted in a dose-dependent inhibition of cell proliferation. This was associated to a rapid and significant increase of ceramide levels and was mimicked by treatments that augment cellular ceramide. Metabolic experiments with radioactive sphingosine, serine, and choline ... More
Quantum dots are powerful multipurpose vital labeling agents in zebrafish embryos.
AuthorsRieger S, Kulkarni RP, Darcy D, Fraser SE, Köster RW
JournalDev Dyn
PubMed ID16110511
Recently, inorganic fluorescent contrast agents composed of semiconductor materials have been introduced to biological imaging approaches. These so-called quantum dots provide unique and promising properties unreached by organic fluorophores, but their use as contrast agents within live organisms has been limited, probably due in part to concerns about their in ... More
A fluorescent lipid analogue can be used to monitor secretory activity and for isolation of mammalian secretion mutants.
AuthorsKtistakis NT, Kao CY, Wang RH, Roth MG
JournalMol Biol Cell
PubMed ID7787242
The use of reporter proteins to study the regulation of secretion has often been complicated by posttranslational processing events that influence the secretion of certain proteins, but are not part of the cellular mechanisms that specifically regulate secretion. This has been a particular limitation for the isolation of mammalian secretion ... More
PDMP blocks brefeldin A-induced retrograde membrane transport from golgi to ER: evidence for involvement of calcium homeostasis and dissociation from sphingolipid metabolism.
AuthorsKok JW, Babia T, Filipeanu CM, Nelemans A, Egea G, Hoekstra D
JournalJ Cell Biol
PubMed ID9660860
In this study, we show that an inhibitor of sphingolipid biosynthesis, D,L-threo-1-phenyl-2- decanoylamino-3-morpholino-1-propanol (PDMP), inhibits brefeldin A (BFA)-induced retrograde membrane transport from Golgi to endoplasmic reticulum (ER). If BFA treatment was combined with or preceded by PDMP administration to cells, disappearance of discrete Golgi structures did not occur. However, when ... More
Internalization and sorting of plasma membrane sphingolipid analogues in differentiating oligodendrocytes.
AuthorsWatanabe R, Asakura K, Rodriguez M, Pagano RE
JournalJ Neurochem
PubMed ID10501180
We studied the formation of early endosomes in differentiating oligodendrocytes and type-2 astrocytes, which are derived from common precursor cells in rat neonates, using fluorescent analogues of lactosylceramide (LacCer) and sulfatide labeled with 4,4-difluoro-5,7-dimethyl-4-bora-3a,4a-diaza-s-indacene++ +-3-pentanoic acid (BODIPY FL C5). These sphingolipid analogues exhibit a concentration-dependent shift in their fluorescence emission ... More
Microtubule independent vesiculation of Golgi membranes and the reassembly of vesicles into Golgi stacks.
AuthorsVeit B, Yucel JK, Malhotra V
JournalJ Cell Biol
PubMed ID8104190
We have recently shown that ilimaquinone (IQ) causes the breakdown of Golgi membranes into small vesicles (VGMs for vesiculated Golgi membranes) and inhibits vesicular protein transport between successive Golgi cisternae (Takizawa et al., 1993). While other intracellular organelles, intermediate filaments, and actin filaments are not affected, we have found that ... More
Defective pH regulation of acidic compartments in human breast cancer cells (MCF-7) is normalized in adriamycin-resistant cells (MCF-7adr).
AuthorsSchindler M, Grabski S, Hoff E, Simon SM
JournalBiochemistry
PubMed ID8608115
Alkalinization of normally acidic intracellular compartments or acidification of a mildly alkaline cytoplasm by biochemical or genetic manipulation has been demonstrated to inhibit both endocytosis and secretion (Tartakoff, 1983a; Cosson et al., 1989; Mellman et al., 1986; Davoust et al., 1987; Cosson et al., 1989; van Deurs et al., 1989; ... More
Targeting of scavenger receptor class B type I by synthetic amphipathic alpha-helical-containing peptides blocks lipopolysaccharide (LPS) uptake and LPS-induced pro-inflammatory cytokine responses in THP-1 monocyte cells.
AuthorsBocharov AV, Baranova IN, Vishnyakova TG, Remaley AT, Csako G, Thomas F, Patterson AP, Eggerman TL
JournalJ Biol Chem
PubMed ID15199068
Human scavenger receptor class B type I, CLA-1, mediates lipopolysaccharide (LPS) binding and internalization (Vishnyakova, T. G., Bocharov, A. V., Baranova, I. N., Chen, Z., Remaley, A. T., Csako, G., Eggerman, T. L., and Patterson, A. P. (2003) J. Biol. Chem. 278, 22771-22780). Because one of the recognition motifs in ... More
A zebrafish sox9 gene required for cartilage morphogenesis.
AuthorsYan YL, Miller CT, Nissen RM, Singer A, Liu D, Kirn A, Draper B, Willoughby J, Morcos PA, Amsterdam A, Chung BC, Westerfield M, Haffter P, Hopkins N, Kimmel C, Postlethwait JH, Nissen R
JournalDevelopment
PubMed ID12397114
The molecular genetic mechanisms of cartilage construction are incompletely understood. Zebrafish embryos homozygous for jellyfish (jef) mutations show craniofacial defects and lack cartilage elements of the neurocranium, pharyngeal arches, and pectoral girdle similar to humans with campomelic dysplasia. We show that two alleles of jef contain mutations in sox9a, one ... More