Subzonal organization of olfactory sensory neurons projecting to distinct glomeruli within the mouse olfactory bulb.
AuthorsLevai O, Breer H, Strotmann J
JournalJ Comp Neurol
PubMed ID12619077
'Olfactory sensory neurons located in the nasal neuroepithelium send their axons directly into the olfactory bulb, where they contact the dendrites of second-order neurons in specialized spherical structures called glomeruli; each sensory neuron projects to a single glomerulus. All neurons expressing the same odorant receptor gene are confined to distinct ... More
Direct membrane protein-DNA interactions required early in nuclear envelope assembly.
AuthorsUlbert S, Platani M, Boue S, Mattaj IW
JournalJ Cell Biol
PubMed ID16717124
'Among the earliest events in postmitotic nuclear envelope (NE) assembly are the interactions between chromatin and the membranes that will fuse to form the NE. It has been proposed that interactions between integral NE proteins and chromatin proteins mediate initial membrane recruitment to chromatin. We show that several transmembrane NE ... More
Antigen-induced translocation of PKC-theta to membrane rafts is required for T cell activation.
AuthorsBi K, Tanaka Y, Coudronniere N, Sugie K, Hong S, van Stipdonk MJ, Altman A
JournalNat Immunol
PubMed ID11376344
'Protein kinase C-theta (PKC-theta) is essential for mature T cell activation; however, the mechanism by which it is recruited to the TCR signaling machinery is unknown. Here we show that T cell stimulation by antibodies or peptide-major histocompatibility complex (MHC) induces translocation of PKC-theta to membrane lipid rafts, which localize ... More
Beta-very low density lipoprotein is sequestered in surface-connected tubules in mouse peritoneal macrophages.
AuthorsMyers JN, Tabas I, Jones NL, Maxfield FR
JournalJ Cell Biol
PubMed ID8253839
'beta-very low density lipoprotein (VLDL) is a large lipoprotein with multiple apoprotein E (apoE) molecules that bind to the LDL receptors on mouse macrophages. Even though they bind to the same receptor, the endocytic processing of beta-VLDL differs from low density lipoprotein (LDL). LDL is rapidly delivered to perinuclear lysosomes ... More
Regulation of C-cadherin function during activin induced morphogenesis of Xenopus animal caps.
AuthorsBrieher WM, Gumbiner BM
JournalJ Cell Biol
PubMed ID8034750
'Treatment of Xenopus animal pole tissue with activin results in the induction of mesodermal cell types and a dramatic elongation of the tissue. The morphogenetic movements involved in the elongation appear similar to those in normal gastrulation, which is driven by cell rearrangement and cell intercalations. We have used this ... More
Carbocyanine postmortem neuronal tracing. Influence of different parameters on tracing distance and combination with immunocytochemistry.
AuthorsLukas JR, Aigner M, Denk M, Heinzl H, Burian M, Mayr R
JournalJ Histochem Cytochem
PubMed ID9671441
'Carbocyanines (DiI, DiA, DiO) are able to travel along membranes by diffusion and therefore have been used as postmortem neuronal tracers in aldehyde-fixed tissues. Surprisingly, detailed data on the influence of different parameters on tracing distances are still missing. This study was carried out to optimize tracing procedures and to ... More
Delivery of ligands from sorting endosomes to late endosomes occurs by maturation of sorting endosomes.
AuthorsDunn KW, Maxfield FR
JournalJ Cell Biol
PubMed ID1560027
'After endocytosis, lysosomally targeted ligands pass through a series of endosomal compartments. The endocytic apparatus that accomplishes this passage may be considered to take one of two forms: (a) a system in which lysosomally targeted ligands pass through preexisting, long-lived early sorting endosomes and are then selectively transported to long-lived ... More
A quantitative study of the segmental distribution of somitic cells in the developing chick limb bud using laser-scanning confocal microscopy.
AuthorsEwan KB, Everett AW
JournalJ Histochem Cytochem
PubMed ID8601694
'Our aim was to map the segmental distribution of somitic cells in the limb before and after the fusion of these cells into myotubes. Somitic cells of the brachial somites were labeled by injection of DiI and DiO into the somitoceles of embryonic Day 2 (E2) embryos. The quantitative distribution ... More
Rapid labeling of neuronal populations by ballistic delivery of fluorescent dyes.
AuthorsGrutzendler J, Tsai J, Gan WB
JournalMethods
PubMed ID12695105
'Particle-mediated ballistic delivery of fluorescent dyes has been recently used to label neuronal populations in a rapid and efficient fashion. Here we describe detailed protocols for this technique as well as recent improvements in its implementation. This technique allows rapid labeling of entire neurons in a Golgi-like manner after membranes ... More
Segmental organization of embryonic diencephalon.
AuthorsFigdor MC, Stern CD
JournalNature
PubMed ID8510755
'The diencephalon is a complex integration centre and intricate relay station of the vertebrate brain. Its development involves the generation of great cellular diversity and neuronal specificity. We report here that it becomes organized in steps, through a stereotyped sequence of neuromeric subdivisions. Diencephalic neuromeres define four cellular domains (D1-D4) ... More
'Recent studies suggest that phosphoinositide kinases may participate in intracellular trafficking or exocytotic events. Because both of these events ultimately require fusion of biological membranes, the susceptibility of membranes containing polyphosphoinositides (PPIs) to divalent cation-induced fusion was investigated. Results of these investigations indicated that artificial liposomes containing PPI or phosphatidic ... More
Fluorescence methods to detect phase boundaries in lipid bilayer mixtures.
'Phase diagrams of lipid mixtures can show several different regions of phase coexistence, which include liquid-disordered, liquid-ordered, and gel phases. Some phase regions are small, and some have sharp boundaries. The identity of the phases, their location in composition space, and the nature of the transitions between the phases are ... More
Release of hydrophobic molecules from polymer micelles into cell membranes revealed by Forster resonance energy transfer imaging.
AuthorsChen H, Kim S, Li L, Wang S, Park K, Cheng JX,
JournalProc Natl Acad Sci U S A
PubMed ID18445654
'It is generally assumed that polymeric micelles, upon administration into the blood stream, carry drug molecules until they are taken up into cells followed by intracellular release. The current work revisits this conventional wisdom. The study using dual-labeled micelles containing fluorescently labeled copolymers and hydrophobic fluorescent probes entrapped in the ... More
Subventricular zone neural stem cells remodel the brain following traumatic injury in adult mice.
AuthorsSalman H, Ghosh P, Kernie SG
JournalJ Neurotrauma
PubMed ID15115603
'Neural stem cells have recently been shown to contribute to the cellular remodeling that occurs following traumatic brain injury (TBI). Potential sources for these stem cells from within the brain include the subventricular zone of the lateral ventricles and the subgranular zone of the dentate gyrus. Using intraventricular injections of ... More
Evaluation of carbocyanine-labeled erythrocytes for microvascular measurements.
'Red blood cells labeled with the carbocyanine dyes, 1,1''-dioctadecyl-3,3,3'',3''-tetramethylindocarbocyanine perchlorate (DiI) and 3,3''-dioctadecyloxacarbocyanine perchlorate (DiO), were evaluated for use in making microvascular measurements in rat small intestine and spinotrapezius muscle. We determined the minimum concentration of each dye which produced near maximal fluorescent intensity and labeled cell fraction. These dyes, ... More
Evaluation of human mast cell-mediated cytotoxicity by DIOC18 target cell labeling in flow cytometry.
AuthorsOzdemir O,
JournalJ Immunol Methods
PubMed ID17188705
'(51)Cr release assay (CRA) is still the standard method to study mast cell (MC)-mediated cytotoxicity in vitro. Non-radioactive methods e.g. MTT, Hoechst 22147 staining, have also been used. Though CRA has the benefit of being reproducible, it has several drawbacks e.g. spontaneous release and radioactivity. The basic strategy of this ... More
Novel Fusogenic Liposomes for Fluorescent Cell Labeling and Membrane Modification.
AuthorsCsisza´r A, Hersch N, Dieluweit S, Biehl R, Merkel R, Hoffmann B,
JournalBioconjug Chem
PubMed ID20184308
'Efficient delivery of biomolecules into membranes of living cells as well as cell surface modifications are major biotechnological challenges. Here, novel liposome systems based on neutral and cationic lipids in combination with lipids modified by aromatic groups are introduced for such applications. The fusion efficiency of these liposome systems was ... More
An acidic microenvironment inhibits antitumoral non-major histocompatibility complex-restricted cytotoxicity: implications for cancer immunotherapy.
AuthorsFischer B, Müller B, Fisch P, Kreutz W
JournalJ Immunother (1997)
PubMed ID10746546
'Local immunosuppression may explain the failure of an effective immune response against solid tumors. Although it is well known that the interstitial pH is significantly lower in solid tumors than in normal tissue, only a few studies in the mouse system have investigated the influence of this acidic milieu on ... More
CR1-mediated ATP Release by Human Red Blood Cells Promotes CR1 Clustering and Modulates the Immune Transfer Process.
AuthorsMelhorn MI, Brodsky AS, Estanislau J, Khoory JA, Illigens B, Hamachi I, Kurishita Y, Fraser AD, Nicholson-Weller A, Dolmatova E, Duffy HS, Ghiran IC,
Journal
PubMed ID24022490
'Humans and other higher primates are unique among mammals in using complement receptor 1 (CR1, CD35) on red blood cells (RBC) to ligate complement-tagged inflammatory particles (immune complexes, apoptotic/necrotic debris, and microbes) in the circulation for quiet transport to the sinusoids of spleen and liver where resident macrophages remove the ... More
Spontaneous regression of advanced cancer: identification of a unique genetically determined, age-dependent trait in mice.
AuthorsCui Z, Willingham MC, Hicks AM, Alexander-Miller MA, Howard TD, Hawkins GA, Miller MS, Weir HM, Du W, DeLong CJ
JournalProc Natl Acad Sci U S A
PubMed ID12724523
'We have established and studied a colony of mice with a unique trait of host resistance to both ascites and solid cancers induced by transplantable cells. One dramatic manifestation of this trait is age-dependent spontaneous regression of advanced cancers. This powerful resistance segregates as a single-locus dominant trait, is independent ... More
Vital imaging and ultrastructural analysis of individual axon terminals labeled by iontophoretic application of lipophilic dye.
AuthorsGan WB, Bishop DL, Turney SG, Lichtman JW
JournalJ Neurosci Methods
PubMed ID10598860
'We describe a method for in vivo confocal fluorescence imaging of synaptic terminals and subsequent electron microscopic reconstructions of the same terminals. By iontophoretically applying lipophilic dye to nerve terminals at a single neuromuscular junction with a sharp microelectrode in living neonatal mice, we were able to quickly label other ... More
Carbocyanine dyes with long alkyl side-chains: broad spectrum inhibitors of mitochondrial electron transport chain activity.
AuthorsAnderson WM, Trgovcich-Zacok D
JournalBiochem Pharmacol
PubMed ID7763312
'Certain indocarbocyanine, thiacarbocyanine, and oxacarbocyanine dyes possessing short alkyl side-chains (one to five carbons) are potent inhibitors of mammalian mitochondrial NADH-ubiquinone reductase (EC 1.6.99.3) activity (Anderson et al., Biochem Pharmacol 41: 677-684, 1991; Anderson et al., Biochem Pharmacol 45: 691-696, 1993; Anderson et al., Biochem Pharmacol 45: 2115-2122, 1993), and ... More
Cadherin-mediated cell sorting not determined by binding or adhesion specificity.
AuthorsNiessen CM, Gumbiner BM
JournalJ Cell Biol
PubMed ID11790800
'Cadherin adhesion molecules play important roles in the establishment of tissue boundaries. Cells expressing different cadherins sort out from each other in cell aggregation assays. To determine the contribution of cadherin binding and adhesion specificity to the sorting process, we examined the adhesion of cells to different purified cadherin proteins. ... More
Occurrence and a possible mechanism of penetration of natural killer cells into K562 target cells during the cytotoxic interaction.
AuthorsRadosevic K, van Leeuwen AM, Segers-Nolten IM, Figdor CG, de Grooth BG, Greve J
JournalCytometry
PubMed ID7587714
'The cytotoxic interaction between cloned human Natural Killer (NK) cells and K562 target cells was studied using confocal laser scanning microscopy (CLSM) and conventional fluorescence microscopy. We observed, using fixed as well as living cells, the occurrence of (pseudo) emperipolesis during the interaction. About 30% of conjugated NK cells penetrated, ... More
Mossy fiber growth and synaptogenesis in rat hippocampal slices in vitro.
AuthorsDailey ME, Buchanan J, Bergles DE, Smith SJ
JournalJ Neurosci
PubMed ID8120613
'Hippocampal slices from early postnatal rat were used to study mossy fiber (MF) growth and synaptogenesis. The ability of MFs to form new giant synapses within isolated tissue slices was established by a series of experiments involving synapsin I immunohistochemistry, electron microscopy, and whole-cell recordings. When hippocampal slices from immature ... More
Requirement for the zebrafish mid-hindbrain boundary in midbrain polarisation, mapping and confinement of the retinotectal projection.
AuthorsPicker A, Brennan C, Reifers F, Clarke JD, Holder N, Brand M
JournalDevelopment
PubMed ID10357940
'The organizer at the midbrain-hindbrain boundary (MHB organizer) has been proposed to induce and polarize the midbrain during development. We investigate the requirement for the MHB organizer in acerebellar mutants, which lack a MHB and cerebellum, but retain a tectum, and are mutant for fgf8, a candidate inducer and polarizer. ... More
Redistribution of cell membrane probes following contraction-induced injury of mouse soleus muscle.
AuthorsWarren GL, Lowe DA, Hayes DA, Farmer MA, Armstrong RB
JournalCell Tissue Res
PubMed ID8565058
'Our aim was to study how mouse skeletal muscle membranes are altered by eccentric and isometric contractions. A fluorescent dialkyl carbocyanine dye (DiOC18(3)) was used to label muscle membranes, and the membranes accessible to the dye were observed by confocal laser scanning microscopy. Experiments were done on normal mouse soleus ... More
Liposome-mediated transfer of macromolecules into flagellated cell envelopes from bacteria.
AuthorsLelkes PI, Klein L, Marikovsky Y, Eisenbach M
JournalBiochemistry
PubMed ID6367823
'We have studied the interaction between flagellated cell envelopes from Escherichia coli and liposomes. Oligolamellar liposomes of ca. 0.45-micron diameter, composed of azolectin, phosphatidylserine, and cholesterol at a molar ratio of 7:1:2, were prepared by freezing and thawing and subsequent extrusion through polycarbonate filters. These liposomes exhibited high entrapment capacity ... More
Endocytosed beta-VLDL and LDL are delivered to different intracellular vesicles in mouse peritoneal macrophages.
AuthorsTabas I, Lim S, Xu XX, Maxfield FR
JournalJ Cell Biol
PubMed ID2391369
'Hypercholesterolemic rabbit beta-VLDL and human LDL are both internalized by mouse peritoneal macrophages by receptor-mediated endocytosis. However, only beta-VLDL (which binds to the cells with a much higher affinity than LDL) markedly stimulates acyl-CoA/cholesterol acyl transferase (ACAT) and induces foam cell formation in these cells. As an initial step to ... More
Functional cis-heterodimers of N- and R-cadherins.
AuthorsShan WS, Tanaka H, Phillips GR, Arndt K, Yoshida M, Colman DR, Shapiro L
JournalJ Cell Biol
PubMed ID10662782
'Classical cadherins form parallel cis-dimers that emanate from a single cell surface. It is thought that the cis-dimeric form is active in cell-cell adhesion, whereas cadherin monomers are likely to be inactive. Currently, cis-dimers have been shown to exist only between cadherins of the same type. Here, we show the ... More
Natural cytotoxic activity of gilthead seabream (Sparus aurata L.) leucocytes. Assessment by flow cytometry and microscopy.
AuthorsCuesta A, Esteban MA, Meseguer J
JournalVet Immunol Immunopathol
PubMed ID10587298
'This paper describes an easy and sensitive flow cytometric assay for assessing the non-specific cytotoxic activity of gilthead seabream (Sparus aurata L.) head-kidney leucocytes against tumor target cells. Concomitantly, the cytotoxic process and the cell types involved were microscopically studied. The assay was based on the consecutive use of two ... More
Noninvasive assessment of angiogenesis by ultrasound and microbubbles targeted to alpha(v)-integrins.
AuthorsLeong-Poi H, Christiansen J, Klibanov AL, Kaul S, Lindner JR
JournalCirculation
PubMed ID12551871
'BACKGROUND: Noninvasive methods for characterizing neovessel formation during angiogenesis are currently lacking. We hypothesized that angiogenesis could be imaged with the use of contrast-enhanced ultrasound (CEU) with microbubbles targeted to alpha(v)-integrins. METHODS AND RESULTS: Microbubbles targeted to alpha(v)-integrins were prepared by conjugating echistatin (MB(E)) or monoclonal antibody against murine alpha(v) ... More
Total internal reflection fluorescence flow cytometry.
AuthorsWang J, Bao N, Paris LL, Geahlen RL, Lu C,
JournalAnal Chem
PubMed ID19007249
'Total internal reflection fluorescence microscopy (TIRFM) has been widely used to explore biological events that are close to the cell membrane by illuminating fluorescent molecules using the evanescent wave. However, TIRFM is typically limited to the examination of a low number of cells, and the results do not reveal potential ... More
Multicolor "DiOlistic" labeling of the nervous system using lipophilic dye combinations.
AuthorsGan WB, Grutzendler J, Wong WT, Wong RO, Lichtman JW
JournalNeuron
PubMed ID10985343
'We describe a technique for rapid labeling of a large number of cells in the nervous system with many different colors. By delivering lipophilic dye-coated particles to neuronal preparations with a "gene gun," individual neurons and glia whose membranes are contacted by the particles are quickly labeled. Using particles that ... More
A versatile flow cytometry-based assay for the determination of short- and long-term natural killer cell activity.
AuthorsJohann S, Blümel G, Lipp M, Förster R
JournalJ Immunol Methods
PubMed ID7561131
'A flow cytometry based method has been developed to assess natural killer (NK) cell activity in both short-term (4 h) and long-term (18 h) NK assays. Target cells were either labeled with PKH-2, c''FDA or D275. Simultaneously, dead cells were identified by counter-staining with the nuclear dye propidium iodide. Using ... More
Dynamic behavior of fluorescent probes in lipid bilayer model membranes.
AuthorsBadley RA, Martin WG, Schneider H
JournalBiochemistry
PubMed ID4683001
Functional expression of a full-length cDNA coding for rat neural cell adhesion molecule L1 mediates homophilic intercellular adhesion and migration of cerebellar neurons.
AuthorsMiura M, Asou H, Kobayashi M, Uyemura K
JournalJ Biol Chem
PubMed ID1587850
'Neural cell adhesion molecule L1 is postulated to be involved in cell-cell interaction, neurite elongation, fasciculation of axons, cell migration, and myelination. To determine the function of L1 directly, we have transfected rat L1 cDNA into mouse fibroblast L cells. Stable transformants expressing L1 showed uniform surface expression of the ... More
Trogocytosis by Entamoeba histolytica contributes to cell killing and tissue invasion.
AuthorsRalston KS, Solga MD, Mackey-Lawrence NM, Somlata, Bhattacharya A, Petri WA,
Journal
PubMed ID24717428
'Entamoeba histolytica is the causative agent of amoebiasis, a potentially fatal diarrhoeal disease in the developing world. The parasite was named '
Growth cone and dendrite dynamics in zebrafish embryos: early events in synaptogenesis imaged in vivo.
AuthorsJontes JD, Buchanan J, Smith SJ
JournalNat Neurosci
PubMed ID10700254
'We used time-lapse fluorescence microscopy to observe the growth of Mauthner cell axons and their postsynaptic targets, the primary motor neurons, in spinal cords of developing zebrafish embryos. Upon reaching successive motor neurons, the Mauthner growth cone paused briefly before continuing along its path. Varicosities formed at regular intervals and ... More
Submillisecond optical reporting of membrane potential in situ using a neuronal tracer dye.
AuthorsBradley J, Luo R, Otis TS, DiGregorio DA,
JournalJ Neurosci
PubMed ID19625510
'A major goal in neuroscience is the development of optical reporters of membrane potential that are easy to use, have limited phototoxicity, and achieve the speed and sensitivity necessary for detection of individual action potentials in single neurons. Here we present a novel, two-component optical approach that attains these goals. ... More
Visualization of galectin-3 oligomerization on the surface of neutrophils and endothelial cells using fluorescence resonance energy transfer.
AuthorsNieminen J, Kuno A, Hirabayashi J, Sato S
JournalJ Biol Chem
PubMed ID17082191
'Galectin-3, a member of the galectin family of carbohydrate binding proteins, is widely expressed, particularly in cells involved in the immune response. Galectin-3 has also been indicated to play a role in various biological activities ranging from cell repression to cell activation and adhesion and has, thus, been recognized as ... More
Clustering of muscle acetylcholine receptors requires motoneurons in live embryos, but not in cell culture.
AuthorsLiu DW, Westerfield M
JournalJ Neurosci
PubMed ID1315852
'Previous culture studies have demonstrated that muscle cells autonomously express and cluster ACh receptors (AChRs) and that contact by neurites induces a reorganization of these clusters. We studied these phenomena in zebrafish embryos where the same cells could be examined in vivo and in vitro, and where contacts between cells ... More
Lipid domains and lipid/protein interactions in biological membranes.
AuthorsTocanne JF, Cézanne L, Lopez A, Piknova B, Schram V, Tournier JF, Welby M
JournalChem Phys Lipids
PubMed ID8001179
'In the fluid mosaic model of membranes, lipids are organized in the form of a bilayer supporting peripheral and integral proteins. This model considers the lipid bilayer as a two-dimensional fluid in which lipids and proteins are free to diffuse. As a direct consequence, both types of molecules would be ... More
Post-formation fluorescent labelling of liposomal membranes. In vivo detection, localisation and kinetics.
AuthorsClaassen E
JournalJ Immunol Methods
PubMed ID1548405
'A fast and simple method for the in vivo/in situ detection of liposomes is described. Utilizing lipophilic carbocyanine dyes, DiI and DiO, yellow (or red) and green fluorescent liposomes can be visualised with routinely available filters. The main advantages of the method are (i) the vesicles can be labelled after ... More
Fluorescence studies on the mechanism of liposome-cell interactions in vitro.
AuthorsSzoka F, Jacobson K, Derzko Z, Papahadjopoulos D
JournalBiochim Biophys Acta
PubMed ID7397162
'Sonicated unilamellar liposomes containing fluorescent lipid analogs or biotinyl phosphatidylethanolamine as a ligand for fluorescein avidin have been used to study the mechanism of interaction of phospholipid vesicles with eucaryotic cells. Microscopy revealed that after short incubations the fluorescence was associated with the cell surface in a punctate as opposed ... More
Lateral diffusion in phospholipid multibilayers measured by fluorescence recovery after photobleaching.
AuthorsWu ES, Jacobson K, Papahadjopoulos D
JournalBiochemistry
PubMed ID901758
'The method of fluorescence recovery after photobleaching has been used to measure the temperature dependence of the lateral diffusion coefficients (D) of two fluorescent lipid analogues in phospholipid multibilayers of various compositions. The probes employed were 3,3-dioctadecyloxocarbocyanine (diO-C18(3) and N-4-nitrobenz-2-oxa-1,3-diazole phosphatidylethanolamine (NBD-PE). In fluid egg phosphatidylcholine multibilayers at 25 degrees ... More
Iterative fractionation of recycling receptors from lysosomally destined ligands in an early sorting endosome.
AuthorsDunn KW, McGraw TE, Maxfield FR
JournalJ Cell Biol
PubMed ID2600137
'To study the fusion and separation of endocytic compartments, we have used digital image analysis to quantify the accumulation of fluorescent ligands in endosomes during continuous endocytosis for periods of 1-20 min. Fluorescently labeled transferrin (Tf) and low density lipoproteins (LDL) were used as markers of recycling receptors and lysosomally ... More
Lateral diffusion and phase separation in two-dimensional solutions of polymerized butadiene lipid in dimyristoylphosphatidylcholine bilayers. A photobleaching and freeze fracture study.
AuthorsGaub H, Sackmann E, Büschl R, Ringsdorf H
JournalBiophys J
PubMed ID6722264
'Mixed vesicles of dimyristoylphosphatidylcholine (DMPC) and a polymerizable lipid containing one diene group per chain are studied by freeze fracture electron microscopy and by the photobleaching (fluorescence recovery after photobleaching) technique. Large thin-walled vesicles of some micron in diameter become more stable after photochemical polymerization. Before polymerization bilayers of the ... More
Embryonic origins of auditory brain-stem nuclei in the chick hindbrain.
AuthorsCramer KS, Fraser SE, Rubel EW
JournalDev Biol
PubMed ID10926755
'The auditory nuclei of the chick brain stem have distinct morphologies and highly specific synaptic connectivity. Nucleus magnocellularis (NM) and nucleus angularis receive tonotopically ordered cochlear input. NM in turn projects tonotopically to nucleus laminaris (NL), maintaining binaural specificity with projections to either dorsal or ventral NL dendrites. NM and ... More
Directional guidance of neuronal migration in the olfactory system by the protein Slit.
AuthorsWu W, Wong K, Chen J, Jiang Z, Dupuis S, Wu JY, Rao Y
JournalNature
PubMed ID10432110
'Although cell migration is crucial for neural development, molecular mechanisms guiding neuronal migration have remained unclear. Here we report that the secreted protein Slit repels neuronal precursors migrating from the anterior subventricular zone in the telencephalon to the olfactory bulb. Our results provide a direct demonstration of a molecular cue ... More
Cytotoxic targeting of F9 teratocarcinoma tumours with anti-ED-B fibronectin scFv antibody modified liposomes.
'We prepared small unilamellar liposomes derivatised with single chain antibody fragments specific for the ED-B domain of B-fibronectin. This extracellular matrix associated protein is expressed around newly forming blood vessels in the vicinity of many types of tumours. The single chain antibody fragments were functionalised by introduction of C-terminal cysteines ... More
Synthesis and characterization of fluorescent Lucifer yellow-lipid conjugates.
AuthorsNothnagel EA
JournalBiochim Biophys Acta
PubMed ID2930788
'The syntheses of fluorescent lipid probes composed of Lucifer yellow dyes linked to either cholesterol or phospholipids are described. The spectral properties of these probes are characterized, and the probes are evaluated for use with model membranes and with live animal and plant cells. Of the probes synthesized, the cholesterol ... More
Disruption of retinal axon ingrowth by ablation of embryonic mouse optic chiasm neurons.
AuthorsSretavan DW, Puré E, Siegel MW, Reichardt LF
JournalScience
PubMed ID7541558
'Mouse retinal ganglion cell axons growing from the eye encounter embryonic neurons at the future site of the optic chiasm. After in vivo ablation of these chiasm neurons with a monoclonal antibody and complement, retinal axons did not cross the midline and stalled at approximately the entry site into the ... More
Labeling with fluorescent carbocyanine dyes of cultured endothelial and smooth muscle cells by growth in dye-containing medium.
AuthorsRagnarson B, Bengtsson L, Haegerstrand A
JournalHistochemistry
PubMed ID1618647
'We describe a method for labeling cultured endothelial cells (ECs) and smooth muscle cells (SMCs) by letting the cells grow for three days in culture medium containing a low concentration of the fluorescent carbocyanine dyes DiI and DiO. We show that good labeling can be obtained with considerably lower concentrations ... More
Fluorescence depolarization measurements on oriented membranes.
AuthorsAdler M, Tritton TR
JournalBiophys J
PubMed ID3165033
'We describe the theory and experimental application of fluorescence depolarization measurements on small molecules bound to oriented phospholipid bilayers. The results yield insight into both the orientation and the rotational motion of fluorophores in a membrane environment. To accomplish this the angular distribution of polarized fluorescence intensities is measured on ... More
Synaptic segregation at the developing neuromuscular junction.
AuthorsGan WB, Lichtman JW
JournalScience
PubMed ID9822385
'Throughout the developing nervous system, competition between axons causes the permanent removal of some synaptic connections. In mouse neuromuscular junctions at birth, terminal branches of different axons are intermingled. However, during the several weeks after birth, these branches progressively segregated into nonoverlapping compartments before the complete withdrawal of all but ... More
The binding of oxidized low density lipoprotein to mouse CD36 is mediated in part by oxidized phospholipids that are associated with both the lipid and protein moieties of the lipoprotein.
AuthorsBoullier A, Gillotte KL, Hörkkö S, Green SR, Friedman P, Dennis EA, Witztum JL, Steinberg D, Quehenberger O
JournalJ Biol Chem
PubMed ID10734051
'There is growing evidence that CD36 has an important physiological function in the uptake of oxidized low density lipoprotein (OxLDL) by macrophages. However, the ligand specificity and the nature of the ligands on OxLDL that mediate the binding to CD36 remain ill defined. Results from recent studies suggested that some ... More
Detection of influenza hemagglutinin interaction with biological membranes by photosensitized activation of [125I]iodonaphthylazide.
AuthorsPak CC, Krumbiegel M, Blumenthal R, Raviv Y
JournalJ Biol Chem
PubMed ID8182068
'Fusion of influenza virus with cells is triggered by a pH-dependent conformational change in the viral envelope protein, hemagglutinin, which results in exposure of the fusion peptide and its insertion into the target membrane. We have investigated the association of hemagglutinin with erythrocyte membranes by photosensitized labeling with [125I]iodonaphthylazide. This ... More
Vertebrate slit, a secreted ligand for the transmembrane protein roundabout, is a repellent for olfactory bulb axons.
'The olfactory bulb plays a central role in olfactory information processing through its connections with both peripheral and cortical structures. Axons projecting from the olfactory bulb to the telencephalon are guided by a repulsive activity in the septum. The molecular nature of the repellent is not known. We report here ... More
Selective inhibition of neurite outgrowth on mature astrocytes by Thy-1 glycoprotein.
AuthorsTiveron MC, Barboni E, Pliego Rivero FB, Gormley AM, Seeley PJ, Grosveld F, Morris R
JournalNature
PubMed ID1346926
'THY-1, the smallest member of the immunoglobulin superfamily, is a major cell-surface component expressed by several tissues. The protein, carbohydrate and gene structures of this molecule are known, yet its function is not. It is highly expressed in nervous tissue, where it appears on virtually all neurons after the cessation ... More
Construction of complex receptive fields in cat primary visual cortex.
AuthorsMartinez LM, Alonso JM
JournalNeuron
PubMed ID11709161
'In primary visual cortex, neurons are classified into simple cells and complex cells based on their response properties. Although the role of these two cell types in vision is still unknown, an attractive hypothesis is that simple cells are necessary to construct complex receptive fields. This hierarchical model puts forward ... More
Pathfinding and error correction by retinal axons: the role of <i>astray/robo2</i>.
AuthorsHutson LD, Chien CB
JournalNeuron
PubMed ID11804569
Dextran amines in neuronal tracing.
AuthorsFritzsch B, Wilm C
JournalTrends Neurosci
PubMed ID1688669
Applications of dye-sensitized photoreactions in neurobiology.
AuthorsSpikes JD
JournalPhotochem Photobiol
PubMed ID1775530
Fluorescence microphotolysis. Diffusion measurements in single cells.
AuthorsPeters R
JournalNaturwissenschaften
PubMed ID6877390
Lateral diffusion of lipids in model and natural membranes.
AuthorsTocanne JF, Dupou-Cézanne L, Lopez A
JournalProg Lipid Res
PubMed ID8022844
Comparative lateral diffusion of fluorescent lipid analogues in phospholipid multibilayers.
AuthorsDerzko Z, Jacobson K
JournalBiochemistry
PubMed ID7470449
Fluorescent dye (DiI) reveals the sensory cells in the lingual epithelium: a confocal laser scanning microscopic study.
AuthorsNagai T
JournalJpn J Physiol
PubMed ID8271492
Fluorescent carbocyanine dye was applied to the glossopharyngeal nerve of axolotls fixed with paraformaldehyde. Three cell types in the lingual epithelium were transneuronally labeled: chemosensory taste receptor cells and presumably mechanosensory basal cells in the taste buds, and epithelial cells with unknown function in the non-taste lingual epithelium. ... More
Lipid lateral diffusion and membrane organization.
AuthorsTocanne JF, Dupou-Cézanne L, Lopez A, Tournier JF
JournalFEBS Lett
PubMed ID2680602
It is shown that investigating the lateral motion of lipids in biological membranes can provide useful information on membrane lateral organization. After labeling membranes with extrinsic or intrinsic lipophilic fluorescent probes, fluorescence recovery after photobleaching experiments strongly suggests that specialized cells like spermatozoa, eggs and epithelia exhibit surface membrane regionalization ... 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
Direct visualisation and quantification of cellular cytotoxicity using two colour flourescence.
AuthorsKroesen BJ, Mesander G, ter Haar JG, The TH, de Leij L
JournalJ Immunol Methods
PubMed ID1431162
A fluorescence method is described for the evaluation of cell death induced by cellular cytolytic activity. A green fluorescent membrane dye, D275, was used to label various target cell lines and propidium iodide (PI) uptake was used to assay cell death. Natural killer (NK), lymphokine activated killer (LAK) as well ... More
We have identified the integrin alpha(v)beta3 as a ligand for mouse gp49B1, thus identifying a new class of ligand for a member of the family of inhibitory immunoreceptors that bear C2-type immunoglobulin-like domains. The specific interaction was shown by both cell-protein and cell-cell binding assays. In addition, we found that ... More
Nanotubular highways for intercellular organelle transport.
AuthorsRustom A, Saffrich R, Markovic I, Walther P, Gerdes HH
JournalScience
PubMed ID14963329
Cell-to-cell communication is a crucial prerequisite for the development and maintenance of multicellular organisms. To date, diverse mechanisms of intercellular exchange of information have been documented, including chemical synapses, gap junctions, and plasmodesmata. Here, we describe highly sensitive nanotubular structures formed de novo between cells that create complex networks. These ... More
Homeotic transformation of rhombomere identity after localized Hoxb1 misexpression.
AuthorsBell E, Wingate RJ, Lumsden A
JournalScience
PubMed ID10381880
Segmentation of the hindbrain and branchial region is a conserved feature of head development, involving the nested expression of Hox genes. Although it is presumed that vertebrate Hox genes function as segment identifiers, responsible for mediating registration between elements of diverse embryonic origin, this assumption has remained untested. To assess ... More
Distinct AAA-ATPase p97 complexes function in discrete steps of nuclear assembly.
Although nuclear envelope (NE) assembly is known to require the GTPase Ran, the membrane fusion machinery involved is uncharacterized. NE assembly involves formation of a reticular network on chromatin, fusion of this network into a closed NE and subsequent expansion. Here we show that p97, an AAA-ATPase previously implicated in ... More
Photophysical studies of 3,3' dioctadecyloxacarbocyanine dye in model biological membranes and different solvents.
AuthorsBhowmik BB, Basu S, Ray D
JournalChem Phys Lipids
PubMed ID11269936
The absorption and fluorescence spectra of 3,3'-dioctadecyloxacarbocyanine [DiOC18(3)], a cationic oxacarbocyanine dye have been studied in aqueous and nonaqueous media containing egg phosphatidylcholine (PC) as well as in different solvents of diverse nature. The results show the evidence of complex formation of the dye in the ground and in the ... More
Exovesicles from human activated dendritic cells fuse with resting dendritic cells, allowing them to present alloantigens.
Dendritic cells (DCs) can release microvesicles, but the latter's numbers, size, and fate are unclear. Fluorescently labeled DCs were visualized by laser-scanning microscopy. Using a Surpass algorithm, we were able to identify and quantify per cell several hundred microvesicles released from the surface of stimulated DCs. We show that most ... More
A custom-made two-photon microscope and deconvolution system.
AuthorsMajewska A, Yiu G, Yuste R
JournalPflugers Arch
PubMed ID11211128
We describe in detail a custom-built two-photon microscope based on a modified confocal scanhead (Olympus Fluoview) and mode-locked Ti:sapphire laser (Coherent Mira 900). This system has internal detectors as well as external whole-field detection and an electrooptical modulator for blanking the beam on flyback and effecting fast changes in excitation ... More
Slit1 and Slit2 cooperate to prevent premature midline crossing of retinal axons in the mouse visual system.
AuthorsPlump AS, Erskine L, Sabatier C, Brose K, Epstein CJ, Goodman CS, Mason CA, Tessier-Lavigne M
JournalNeuron
PubMed ID11804570
During development, retinal ganglion cell (RGC) axons either cross or avoid the midline at the optic chiasm. In Drosophila, the Slit protein regulates midline axon crossing through repulsion. To determine the role of Slit proteins in RGC axon guidance, we disrupted Slit1 and Slit2, two of three known mouse Slit ... More
Restricted movement of lipid and aqueous dyes through pores formed by influenza hemagglutinin during cell fusion.
AuthorsZimmerberg J, Blumenthal R, Sarkar DP, Curran M, Morris SJ
JournalJ Cell Biol
PubMed ID7806567
The fusion of cells by influenza hemagglutinin (HA) is the best characterized example of protein-mediated membrane fusion. In simultaneous measurements of pairs of assays for fusion, we determined the order of detectable events during fusion. Fusion pore formation in HA-triggered cell-cell fusion was first detected by changes in cell membrane ... More
Selective electrofusion of conjugated cells in flow.
AuthorsBakker Schut TC, Kraan YM, Barlag W, de Leij L, de Grooth BG, Greve J
JournalBiophys J
PubMed ID8218887
Using a modified flow cytometer we have induced electrofusion of K562 and L1210 cells in flow. The two cell types are stained with two different fluorescent membrane probes, DiO and DiI, to facilitate optical recognition, and then coupled through an avidin-biotin bridge. In the flow cytometer, the hydrodynamically focused cells ... More
A fate map of the epiblast of the early chick embryo.
AuthorsHatada Y, Stern CD
JournalDevelopment
PubMed ID7607078
We have used carbocyanine dyes (DiI and DiO) to generate fate maps for the epiblast layer of the chick embryo between stage X and the early primitive streak stage (stages 2-3). The overall distribution of presumptive cell types in these maps is similar to that described for other laboratory species ... More
Ultrasound radiation force modulates ligand availability on targeted contrast agents.
Radiation force produced by low-amplitude ultrasound at clinically relevant frequencies remotely translates freely flowing microbubble ultrasound contrast agents over distances up to centimeters from the luminal space to the vessel wall in order to enhance ligand-receptor contact in targeting applications. The question arises as to how the microbubble shell might ... More
Diolistics: incorporating fluorescent dyes into biological samples using a gene gun.
AuthorsO'Brien JA, Lummis SC,
JournalTrends Biotechnol
PubMed ID17945370
The hand-held gene gun provides a rapid and efficient method of incorporating fluorescent dyes into cells, a technique that is becoming known as diolistics. Transporting fluorescent dyes into cells has, in the past, used predominantly injection or chemical methods. The use of the gene gun, combined with the new generation ... More
Lipoplex formulation of superior efficacy exhibits high surface activity and fusogenicity, and readily releases DNA.
AuthorsKoynova R, Tarahovsky YS, Wang L, MacDonald RC,
JournalBiochim Biophys Acta
PubMed ID17156744
Lipoplexes containing a mixture of cationic phospholipids dioleoylethylphosphatidylcholine (EDOPC) and dilauroylethylphosphatidylcholine (EDLPC) are known to be far more efficient agents in transfection of cultured primary endothelial cells than are lipoplexes containing either lipid alone. The large magnitude of the synergy permits comparison of the physical and physico-chemical properties of lipoplexes ... More
How curved membranes recruit amphipathic helices and protein anchoring motifs.
Lipids and several specialized proteins are thought to be able to sense the curvature of membranes (MC). Here we used quantitative fluorescence microscopy to measure curvature-selective binding of amphipathic motifs on single liposomes 50-700 nm in diameter. Our results revealed that sensing is predominantly mediated by a higher density of ... More
Hydrophobic inactivation of influenza viruses confers preservation of viral structure with enhanced immunogenicity.
AuthorsRaviv Y, Blumenthal R, Tompkins SM, Humberd J, Hogan RJ, Viard M,
JournalJ Virol
PubMed ID18305038
The use of inactivated influenza virus for the development of vaccines with broad heterosubtypic protection requires selective inactivation techniques that eliminate viral infectivity while preserving structural integrity. Here we tested if a hydrophobic inactivation approach reported for retroviruses could be applied to the influenza virus. By this approach, the transmembrane ... More
Rapid triple-labeling method combining in situ hybridization and double immunocytochemistry.
AuthorsLopez-Sanchez C, Garcia-Martinez V, Lawson A, Chapman SC, Schoenwolf GC,
JournalDev Dyn
PubMed ID15162509
A new, rapid method is described for combining in situ hybridization and immunocytochemistry to define cell populations and to map three-dimensional movements of groups of labeled cells within developing chick embryos. The method allows fluorescently labeled cells to be followed in living embryos and subsequently detected as a permanent reaction ... More
Suitability of various membrane lipophilic probes for the detection of trogocytosis by flow cytometry.
AuthorsDaubeuf S, Bordier C, Hudrisier D, Joly E,
JournalCytometry A
PubMed ID19051238
Trogocytosis is a recently discovered phenomenon whereby lymphocytes capture fragments of the plasma membrane from antigen presenting cells (APCs). Using APCs labeled with widely used fluorescent lipophilic probes, we previously described a trogocytosis analysis protocol (TRAP) useful to understand the mechanisms and biological consequences of this process and to identify ... More
A proposal for a coordinated effort for the determination of brainwide neuroanatomical connectivity in model organisms at a mesoscopic scale.
AuthorsBohland JW, Wu C, Barbas H, Bokil H, Bota M, Breiter HC, Cline HT, Doyle JC, Freed PJ, Greenspan RJ, Haber SN, Hawrylycz M, Herrera DG, Hilgetag CC, Huang ZJ, Jones A, Jones EG, Karten HJ, Kleinfeld D, Kötter R, Lester HA, Lin JM, Mensh BD, Mikula S, Panksepp J, Price JL, Safdieh J, Saper CB, Schiff ND, Schmahmann JD, Stillman BW, Svoboda K, Swanson LW, Toga AW, Van Essen DC, Watson JD, Mitra PP,
JournalPLoS Comput Biol
PubMed ID19325892
In this era of complete genomes, our knowledge of neuroanatomical circuitry remains surprisingly sparse. Such knowledge is critical, however, for both basic and clinical research into brain function. Here we advocate for a concerted effort to fill this gap, through systematic, experimental mapping of neural circuits at a mesoscopic scale ... More
Deoxycholate promotes survival of breast cancer cells by reducing the level of pro-apoptotic ceramide.
AuthorsKrishnamurthy K, Wang G, Rokhfeld D, Bieberich E,
JournalBreast Cancer Res
PubMed ID19087284
INTRODUCTION: At physiologic concentration in serum, the bile acid sodium deoxycholate (DC) induces survival and migration of breast cancer cells. Here we provide evidence of a novel mechanism by which DC reduces apoptosis that is induced by the sphingolipid ceramide in breast cancer cells. METHODS: Murine mammacarcinoma 4T1 cells were ... More
Dual-color fluorescence-burst analysis to probe protein efflux through the mechanosensitive channel MscL.
Authorsvan den Bogaart G, Krasnikov V, Poolman B
JournalBiophys J
PubMed ID17142294
The mechanosensitive channel protein of large conductance, MscL, from Escherichia coli has been implicated in protein efflux, but the passage of proteins through the channel has never been demonstrated. We used dual-color fluorescence-burst analysis to evaluate the efflux of fluorescent labeled compounds through MscL. The method correlates the fluctuations in ... More
Microfluidic device for combinatorial fusion of liposomes and cells.
AuthorsStrömberg A, Karlsson A, Ryttsén F, Davidson M, Chiu DT, Orwar O
JournalAnal Chem
PubMed ID11195496
We describe an electrofusion-based technique for combinatorial synthesis of individual liposomes. A prototype device with containers for liposomes of different compositions and a fusion container was constructed. The sample containers had fluid contact with the fusion container through microchannels. Optical trapping was used to transport individual liposomes and cells through ... More
Impurity controlled phase transitions of phospholipid monolayers.
AuthorsLösche M, Möhwald H
JournalEur Biophys J
PubMed ID6468343
The phase diagram of monolayers of L-alpha-dimyristoyl phosphatidic acid has been studied by fluorescence microscopy. For pressures corresponding to the nearly horizontal slope in the pressure area diagram the growth of crystalline platelets can be observed. They are of dendritic nature; their sizes can be controlled via pressure, compression speed, ... More
Applied electric fields accelerate the diffusion rate and increase the diffusion distance of DiI in fixed tissue.
AuthorsSwift MJ, Crago PE, Grill WM
JournalJ Neurosci Methods
PubMed ID15585299
Lipophilic carbocyanine dyes are effective neuronal tracers in fixed tissue. However, their application has been limited by the slow diffusion, short tracing distances, and long durations of incubation in fixed tissue. We used applied dc electric fields, that exerted forces on the cationic dyes, to increase the diffusion velocity and ... More
Dil and diO: versatile fluorescent dyes for neuronal labelling and pathway tracing.
AuthorsHonig MG, Hume RI
JournalTrends Neurosci
PubMed ID2480673
The fluorescent carbocyanine dyes dil and diO have an extensive history of use in cell biology, but their use as neuronal tracers is relatively recent. We found in 1985 that these molecules were excellent retrograde and anterograde tracers in the developing nervous system. We went on to show that these ... More
Rapid flow cytometric assay for the assessment of natural killer cell activity.
A new assay using flow cytometry has been established to assess natural killer (NK) lytic activity with common bench top instrumentation. This assay uses a cyanine membrane dye to stain live K562 target cells and an iodide nuclear dye to evaluate dead cells, and provides a method of reliably separating ... More
Restriction in IgM expression--VI. Affinity analysis of monoclonal anti-dansyl antibodies.
AuthorsFan ST, Karush F
JournalMol Immunol
PubMed ID6513933
Affinity restriction in the IgM response to the 5-dimethylaminonaphthalene-1-sulfonyl (dansyl) group was studied with 56 monoclonal IgG and IgM affinity-purified antibodies. These were generated by immunization with dansyl-Ficoll or dansyl-B gamma G. Association constants for N epsilon-dansyl-lysine were determined by fluorescence titration based on resonance energy transfer. The antibodies were ... More
Fluorescence spectroscopy of an oriented model membrane.
AuthorsYguerabide J, Stryer L
JournalProc Natl Acad Sci U S A
PubMed ID5288369
We have devised a simple method that makes it feasible to apply fluorescence techniques to lipid bilayer membranes to elucidate aspects of their structure and dynamics. Fluorescence excitation, emission, and polarization spectra were obtained from a single spherical bilayer membrane consisting of oxidized cholesterol and fluorescent probe. The emission transition ... More
Electroinjection of colloid particles and biopolymers into single unilamellar liposomes and cells for bioanalytical applications.
AuthorsKarlsson M, Nolkrantz K, Davidson MJ, Strömberg A, Ryttsén F, Akerman B, Orwar O
JournalAnal Chem
PubMed ID11128948
A combined electroporation and pressure-driven microinjection method for efficient loading of biopolymers and colloidal particles into single-cell-sized unilamellar liposomes was developed. Single liposomes were positioned between a approximately 2-microm tip diameter solute-filled glass micropipet, equipped with a Pt electrode, and a 5-microm-diameter carbon fiber electrode. A transient, 1-10 ms, rectangular ... More