The mitochondrial death/life regulator in apoptosis and necrosis.
AuthorsKroemer G,Dallaporta B,Resche-Rigon M
JournalAnnual review of physiology
PubMed ID9558479
Both physiological cell death (apoptosis) and, in some cases, accidental cell death (necrosis) involve a two-step process. At a first level, numerous physiological and some pathological stimuli trigger an increase in mitochondrial membrane permeability. The mitochondria release apoptogenic factors through the outer membrane and dissipate the electrochemical gradient of the ... More
Alcohol-induced thymocyte apoptosis is accompanied by impaired mitochondrial function.
AuthorsWang JF,Spitzer JJ
JournalAlcohol (Fayetteville, N.Y.)
PubMed ID9014030
This study examines the effects of chronic alcohol consumption on thymic apoptosis with or without pretreatment with E. coli lipopolysaccharide (LPS). Apoptotic cell death of thymocytes was monitored by DNA fragments in gel electrophoresis and the appearance of apoptotic cells by flow cytometry. Changes in mitochondrial membrane potential (MMP), as ... More
The dynamin-related GTPase, Dnm1p, controls mitochondrial morphology in yeast.
AuthorsOtsuga D, Keegan BR, Brisch E, Thatcher JW, Hermann GJ, Bleazard W, Shaw JM
JournalJ Cell Biol
PubMed ID9786946
'The Saccharomyces cerevisiae Dnm1 protein is structurally related to dynamin, a GTPase required for membrane scission during endocytosis. Here we show that Dnm1p is essential for the maintenance of mitochondrial morphology. Disruption of the DNM1 gene causes the wild-type network of tubular mitochondrial membranes to collapse to one side of ... More
Involvement of endonuclease G in nucleosomal DNA fragmentation under sustained endogenous oxidative stress.
AuthorsIshihara Y, Shimamoto N
JournalJ Biol Chem
PubMed ID16407272
'We have previously shown that inhibition of catalase and glutathione peroxidase activities by 3-amino-1,2,4-triazole (ATZ) and mercaptosuccinic acid (MS), respectively, in rat primary hepatocytes caused sustained endogenous oxidative stress and apoptotic cell death without caspase-3 activation. In this study, we investigated the mechanism of this apoptotic cell death in terms ... More
KIF1B, a novel microtubule plus end-directed monomeric motor protein for transport of mitochondria.
AuthorsNangaku M, Sato-Yoshitake R, Okada Y, Noda Y, Takemura R, Yamazaki H, Hirokawa N
JournalCell
PubMed ID7528108
'To further elucidate the mechanism of organelle transport, we cloned a novel member of the mouse kinesin superfamily, KIF1B. This N-terminal-type motor protein is expressed ubiquitously in various kinds of tissues. In situ hybridization revealed that KIF1B is expressed abundantly in differentiated nerve cells. Interestingly, K1F1B works as a monomer, ... More
Nitric oxide induces tyrosine nitration and release of cytochrome c preceding an increase of mitochondrial transmembrane potential in macrophages.
AuthorsHortelano S, Alvarez AM, Boscá L
JournalFASEB J
PubMed ID10593878
'Treatment of elicited peritoneal macrophages or the macrophage cell line RAW 264.7 with high concentrations of nitric oxide donors is followed by apoptotic cell death. Analysis of the changes in the mitochondrial transmembrane potential (DeltaPsi(m)) with specific fluorescent probes showed a rapid and persistent increase of DeltaPsi(m), a potential that ... More
HA95 and LAP2 beta mediate a novel chromatin-nuclear envelope interaction implicated in initiation of DNA replication.
AuthorsMartins S, Eikvar S, Furukawa K, Collas P
JournalJ Cell Biol
PubMed ID12538639
'HA95 is a chromatin-associated protein that interfaces the nuclear envelope (NE) and chromatin. We report an interaction between HA95 and the inner nuclear membrane protein lamina-associated polypeptide (LAP) 2 beta, and a role of this association in initiation of DNA replication. Precipitation of GST-LAP2 beta fusion proteins and overlays of ... 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
Apoptosis in tumour cells photosensitized with Rose Bengal acetate is induced by multiple organelle photodamage.
AuthorsSoldani C, Croce AC, Bottone MG, Fraschini A, Biggiogera M, Bottiroli G, Pellicciari C,
JournalHistochem Cell Biol
PubMed ID17849139
'Rose Bengal (RB) is a very efficient photosensitizer which undergoes inactivation of its photophysical and photochemical properties upon addition of a quencher group-i.e. acetate-to the xanthene rings. The resulting RB acetate (RB-Ac) derivative behaves as a fluorogenic substrate: it easily enters the cells where the native photoactive molecule is restored ... More
Lamin activity is essential for nuclear envelope assembly in a Drosophila embryo cell-free extract.
AuthorsUlitzur N, Harel A, Feinstein N, Gruenbaum Y
JournalJ Cell Biol
PubMed ID1527167
'The role of the Drosophila lamin protein in nuclear envelope assembly was studied using a Drosophila in vitro assembly system that reconstitutes nuclei from added sperm chromatin or naked DNA. Upon incubation of the embryonic assembly extract with anti-Drosophila lamin antibodies, the attachment of nuclear membrane vesicles to chromatin surface ... More
Cyclopentenone prostaglandins as potential inducers of intracellular oxidative stress.
AuthorsKondo M, Oya-Ito T, Kumagai T, Osawa T, Uchida K
JournalJ Biol Chem
PubMed ID11278531
'In the present study, we find that cyclopentenone prostaglandins (PGs) of the J(2) series, naturally occurring derivatives of PGD(2), are potential inducers of intracellular oxidative stress that mediates cell degeneration. Based on an extensive screening of diverse chemical agents on induction of intracellular production of reactive oxygen species (ROS), we ... More
Reorganization of cytoplasmic structures during cell fusion.
AuthorsZheng QA, Chang DC
JournalJ Cell Sci
PubMed ID1808197
'In order to provide a better understanding of the dynamic process of cell fusion, we studied the reorganization of cytoplasmic structures in electro-fused CV-1 cells. Using fluorescence microscopy and double staining methods, we examined correlations between the structural patterns of the major cytoskeletal proteins (microtubules, actin and vimentin intermediate filaments) ... More
Kinesin-mediated organelle translocation revealed by specific cellular manipulations.
AuthorsFeiguin F, Ferreira A, Kosik KS, Caceres A
JournalJ Cell Biol
PubMed ID7962067
'The distribution of membrane-bound organelles was studied in cultured hippocampal neurons after antisense oligonucleotide suppression of the kinesin-heavy chain (KHC). We observed reduced 3,3''-dihexyloxacarbocyanine iodide (DiOC6(3)) fluorescent staining in neurites and growth cones. In astrocytes, KHC suppression results in the disappearance of the DiOC6(3)-positive reticular network from the cell periphery, ... More
Methods for assessing autophagy and autophagic cell death.
AuthorsTasdemir E, Galluzzi L, Maiuri MC, Criollo A, Vitale I, Hangen E, Modjtahedi N, Kroemer G,
JournalMethods Mol Biol
PubMed ID18425442
'Autophagic (or type 2) cell death is characterized by the massive accumulation of autophagic vacuoles (autophagosomes) in the cytoplasm of cells that lack signs of apoptosis (type 1 cell death). Here we detail and critically assess a series of methods to promote and inhibit autophagy via pharmacological and genetic manipulations. ... More
Rapid movements of vimentin on microtubule tracks: kinesin-dependent assembly of intermediate filament networks.
AuthorsPrahlad V, Yoon M, Moir RD, Vale RD, Goldman RD
JournalJ Cell Biol
PubMed ID9763428
'The assembly and maintenance of an extended intermediate filament (IF) network in fibroblasts requires microtubule (MT) integrity. Using a green fluorescent protein-vimentin construct, and spreading BHK-21 cells as a model system to study IF-MT interactions, we have discovered a novel mechanism involved in the assembly of the vimentin IF cytoskeleton. ... More
Early transmembrane potential changes of lymphocytes in mixed lymphocyte cultures as detected by flow-cytometry.
AuthorsNerl C, Valet G, Schendel DJ, Wank R
JournalNaturwissenschaften
PubMed ID7110362
Non-linear relationship between fluorescence and membrane potential.
AuthorsBurchkhardt G
JournalBiochim Biophys Acta
PubMed ID560205
A trypsin-sensitive receptor on membrane vesicles is required for nuclear envelope formation in vitro.
AuthorsWilson KL, Newport J
JournalJ Cell Biol
PubMed ID3392106
'The reformation of functioning organelles at the end of mitosis presents a problem in vesicle targeting. Using extracts made from Xenopus laevis frog eggs, we have studied in vitro the vesicles that reform the nuclear envelope. In the in vitro assay, nuclear envelope growth is linear with time. Furthermore, the ... 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
Prevention of anti-IgM-induced apoptosis accompanying G1 arrest in B lymphoma cells overexpressing dominant-negative mutant form of c-Jun N-terminal kinase 1.
AuthorsTakada E, Toyota H, Suzuki J, Mizuguchi J
JournalJ Immunol
PubMed ID11160206
'A family of mitogen-activated protein (MAP) kinases comprising the extracellular signal-regulated kinases (ERKs), c-Jun N-terminal kinases (JNKs), and p38 MAP kinases are involved in proliferation and apoptosis. However, there are some arguments concerning the role of these kinases in Ag-induced B cell apoptosis. Two of the B lymphoma cell lines ... More
An early oxygen-dependent step is required for dexamethasone-induced apoptosis of immature mouse thymocytes.
'The roles of oxygen and reactive oxygen intermediates in apoptosis are unclear at present. Although oxygen and reactive oxygen intermediates are not required for the execution of apoptosis, oxygen may be involved in at least some forms of apoptosis. In this study we show that dexamethasone (Dex)-induced apoptosis of immature ... More
Modulation of Mg2+ efflux from rat ventricular myocytes studied with the fluorescent indicator furaptra.
AuthorsTursun P, Tashiro M, Konishi M
JournalBiophys J
PubMed ID15626700
'The fluorescent Mg(2+) indicator furaptra (mag-fura-2) was introduced into single ventricular myocytes by incubation with its acetoxy-methyl ester form. The ratio of furaptra's fluorescence intensity at 382 and 350 nm was used to estimate the apparent cytoplasmic [Mg(2+)] ([Mg(2+)](i)). In Ca(2+)-free extracellular conditions (0.1 mM EGTA) at 25 degrees C, ... More
Activation of mitochondria and release of mitochondrial apoptogenic factors by betulinic acid.
AuthorsFulda S, Scaffidi C, Susin SA, Krammer PH, Kroemer G, Peter ME, Debatin KM
JournalJ Biol Chem
PubMed ID9852046
'Different classes of anticancer drugs may trigger apoptosis by acting on different subcellular targets and by activating distinct signaling pathways. Here, we report that betulinic acid (BetA) is a prototype cytotoxic agent that triggers apoptosis by a direct effect on mitochondria. In isolated mitochondria, BetA directly induces loss of transmembrane ... More
Nitric oxide-dependent mitochondrial biogenesis generates Ca2+ signaling profile of lupus T cells.
AuthorsNagy G, Barcza M, Gonchoroff N, Phillips PE, Perl A
JournalJ Immunol
PubMed ID15356113
'Abnormal T cell activation and cell death underlie the pathology of systemic lupus erythematosus. Although mitochondrial hyperpolarization (MHP) represents an early and reversible checkpoint of T cell activation and apoptosis, lupus T cells exhibit persistent MHP. NO has recently been recognized as a key signal of mitochondrial biogenesis and mediator ... More
Dual control of replication timing. Stochastic onset but programmed completion of mammalian chromosome duplication.
AuthorsAnglana M, Debatisse M
JournalJ Biol Chem
PubMed ID11454865
'In mammalian cells, DNA replication proceeds according to a precise temporal order during the S phase, but how this program is controlled remains poorly understood. We analyzed the replication-dependent bromodeoxyuridine banding of chromosomes in Chinese hamster cells treated with the spindle poison nocodazole. In these cells, nocodazole induces a transient ... More
The NF-kappa B cascade is important in Bcl-xL expression and for the anti-apoptotic effects of the CD28 receptor in primary human CD4+ lymphocytes.
AuthorsKhoshnan A, Tindell C, Laux I, Bae D, Bennett B, Nel AE
JournalJ Immunol
PubMed ID10925251
'We explored the role of the NF-kappa B pathway in the survival of primary human CD4+ T lymphocytes during CD28 costimulation. Transduction of proliferating CD4+ T cells with a tetracycline-regulated retrovirus encoding for a dominant-interfering, degradation-resistant I-kappaBalpha (inhibitor of kappa B alpha factor) mutant induced apoptosis. Using DNA arrays, we ... More
Peroxisomal membrane ascorbate peroxidase is sorted to a membranous network that resembles a subdomain of the endoplasmic reticulum.
'The peroxisomal isoform of ascorbate peroxidase (APX) is a novel membrane isoform that functions in the regeneration of NAD(+) and protection against toxic reactive oxygen species. The intracellular localization and sorting of peroxisomal APX were examined both in vivo and in vitro. Epitope-tagged peroxisomal APX, which was expressed transiently in ... More
Evaluation of fluorescent dyes for the detection of mitochondrial membrane potential changes in cultured cardiomyocytes.
AuthorsMathur A, Hong Y, Kemp BK, Barrientos AA, Erusalimsky JD
JournalCardiovasc Res
PubMed ID10727661
'OBJECTIVE: Maintenance of the mitochondrial membrane potential (Deltapsim) is fundamental for the normal performance and survival of cells such as cardiomyocytes, that have a high energy requirement. Measurement of Deltapsim is therefore essential in order to develop an understanding of the molecular mechanisms controlling cardiomyocyte function. Here we have evaluated ... More
Regulation of the Fas death pathway by FLICE-inhibitory protein in primary human B cells.
AuthorsHennino A, Berard M, Casamayor-Pallejà M, Krammer PH, Defrance T
JournalJ Immunol
PubMed ID10975811
'The Fas/Fas ligand (L) system plays an important role in the maintenance of peripheral B cell tolerance and the prevention of misguided T cell help. CD40-derived signals are required to induce Fas expression on virgin B cells and to promote their susceptibility to Fas-mediated apoptosis. In the current study, we ... More
Role of the mitochondrial membrane permeability transition (MPT) in rotenone-induced apoptosis in liver cells.
AuthorsIsenberg JS, Klaunig JE
JournalToxicol Sci
PubMed ID10696782
'Rotenone inhibits spontaneously and chemically induced hepatic tumorigenesis in rodents through the induction of apoptosis. However, the mechanism for the induction of apoptosis by rotenone has not been defined. Mitochondrial dysfunction, in particular the induction of the mitochondrial membrane permeability transition (MPT), has been implicated in the cascade of events ... More
The yeast gene, MDM20, is necessary for mitochondrial inheritance and organization of the actin cytoskeleton.
AuthorsHermann GJ, King EJ, Shaw JM
JournalJ Cell Biol
PubMed ID9105043
'In Saccharomyces cerevisiae, the growing bud inherits a portion of the mitochondrial network from the mother cell soon after it emerges. Although this polarized transport of mitochondria is thought to require functions of the cytoskeleton, there are conflicting reports concerning the nature of the cytoskeletal element involved. Here we report ... 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
Commitment to apoptosis is associated with changes in mitochondrial biogenesis and activity in cell lines conditionally immortalized with simian virus 40.
AuthorsVayssiere JL, Petit PX, Risler Y, Mignotte B
JournalProc Natl Acad Sci U S A
PubMed ID7972136
'Rodent embryo cells immortalized with temperature-sensitive mutants of simian virus 40 large tumor (T) antigen have a proliferative potential that depends on temperature. At the restrictive temperature, heat-inactivation of large T antigen causes p53 release, growth arrest, and cell death. Morphological and molecular analysis indicate that the induced cell death ... More
Determination of intracellular organelles implicated in daunorubicin cytoplasmic sequestration in multidrug-resistant MCF-7 cells using fluorescence microscopy image analysis.
AuthorsBour-Dill C, Gramain MP, Merlin JL, Marchal S, Guillemin F
JournalCytometry
PubMed ID10655559
'BACKGROUND: Anthracycline resistance is known to be mediated by P-glycoprotein (P-gp) or multidrug-resistance related protein (MRP) as well as intracellular sequestration of drugs. METHODS: The resistance phenotype of doxorubicin-selected MCF-7(DXR) human breast adenocarcinoma cell line was characterized by cellular and nuclear daunorubicin efflux, P-gp and MRP expression and apoptosis induction. ... More
Elemental propagation of calcium signals in response-specific patterns determined by environmental stimulus strength.
AuthorsGoddard H, Manison NF, Tomos D, Brownlee C
JournalProc Natl Acad Sci U S A
PubMed ID10677557
'Plant cells can respond qualitatively and quantitatively to a wide range of environmental signals. Ca(2+) is used as an intracellular signal for volume regulation in response to external osmotic changes. We show here that the spatiotemporal patterns of hypo-osmotically induced Ca(2+) signals vary dramatically with stimulus strength in embryonic cells ... More
Effect of membrane potential on the cellular uptake of 2-N-methyl-ellipticinium by L1210 cells.
AuthorsCharcosset JY, Jacquemin-Sablon A, Le Pecq JB
JournalBiochem Pharmacol
PubMed ID6466349
'Some quaternary ammonium derivatives of ellipticine are active antitumor drugs on both experimental and human tumors. Because of their positive charge, the cellular uptake of these molecules is expected to be influenced by the electric membrane potential. Experimental variations of the potential were produced by changing the external potassium concentration ... More
TPX2 is required for postmitotic nuclear assembly in cell-free Xenopus laevis egg extracts.
AuthorsO'Brien LL, Wiese C
JournalJ Cell Biol
PubMed ID16735579
'Cell division in many metazoa is accompanied by the disassembly of the nuclear envelope and the assembly of the mitotic spindle. These dramatic structural rearrangements are reversed after mitosis, when the mitotic spindle is dismantled and the nuclear envelope reassembles. The targeting protein for XKlp2 (TPX2) plays important roles in ... More
Targeting of membranes to sea urchin sperm chromatin is mediated by a lamin B receptor-like integral membrane protein.
AuthorsCollas P, Courvalin JC, Poccia D
JournalJ Cell Biol
PubMed ID8991085
'We have identified an integral membrane protein of sea urchin gametes with an apparent molecular mass of 56 kD that cross-reacts with an antibody against the nucleoplasmic NH2-terminal domain of human lamin B receptor (LBR). In mature sperm, p56 is located at the tip and base of the nucleus from ... More
Determination of differential cell counts in feline bone marrow by use of flow cytometry.
AuthorsWeiss DJ
JournalAm J Vet Res
PubMed ID11327450
'OBJECTIVE: To evaluate the potential usefulness of 2 flow cytometric methods for determination of differential cell counts in feline bone marrow. SAMPLE POPULATION: 10 bone marrow specimens from client-owned cats. PROCEDURE: Bone marrow specimens were stained with 3,3''-dihexyloxacarbocyanine iodide (DiOC6) and evaluated by use of flow cytometry. Differential counts were ... More
Atovaquone, a broad spectrum antiparasitic drug, collapses mitochondrial membrane potential in a malarial parasite.
AuthorsSrivastava IK, Rottenberg H, Vaidya AB
JournalJ Biol Chem
PubMed ID9020100
'At present, approaches to studying mitochondrial functions in malarial parasites are quite limited because of the technical difficulties in isolating functional mitochondria in sufficient quantity and purity. We have developed a flow cytometric assay as an alternate means to study mitochondrial functions in intact erythrocytes infected with Plasmodium yoelii, a ... More
Increased perinuclear Ca2+ activity evoked by metabotropic glutamate receptor activation in rat hippocampal neurones.
AuthorsPhenna S, Jane SD, Chad JE
JournalJ Physiol
PubMed ID7562631
'1. The effect of metabotropic glutamate receptor activation on intracellular Ca2+ activity (alpha Cai) of rat hippocampal pyramidal neurones in vitro was examined using ratiometric confocal laser scanning microscopy with the Ca(2+)-sensitive fluorescent probe indo-1 AM. 2. Metabotropic receptors were selectively activated with 1S,3R-1-aminocyclopentane-1,3-dicarboxylic acid (1S,3R-ACPD; 100 microM) in the ... More
Mechanism of calcium transport stimulated by chlorothiazide in mouse distal convoluted tubule cells.
AuthorsGesek FA, Friedman PA
JournalJ Clin Invest
PubMed ID1322939
'Thiazide diuretics inhibit Na+ and stimulate Ca2+ absorption in renal distal convoluted tubules. Experiments were performed on immortalized mouse distal convoluted tubule (MDCT) cells to determine the mechanism underlying the dissociation of sodium from calcium transport and the stimulation of calcium absorption induced by thiazide diuretics. Control rates of 22Na+ ... More
Organelle-cytoskeletal interactions: actin mutations inhibit meiosis-dependent mitochondrial rearrangement in the budding yeast Saccharomyces cerevisiae.
AuthorsSmith MG, Simon VR, O'Sullivan H, Pon LA
JournalMol Biol Cell
PubMed ID8573793
'During early stages of meiosis I, yeast mitochondria fuse to form a single continuous thread. Thereafter, portions of the mitochondrial thread are equally distributed to daughter cells. Using time-lapse fluorescence microscopy and a membrane potential sensing dye, mitochondria are resolved as small particles at the cell periphery in pre-meiotic, living ... More
Membrane potential changes after infection of monocytes by Toxoplasma gondii.
AuthorsBouchot A, Millot JM, Charpentier S, Bonhomme A, Villena I, Aubert D, Pinon JM
JournalInt J Parasitol
PubMed ID11429176
'Membrane potential changes in host cell plasma membrane were analyzed and the parasitophorous vacuole membrane (PVM) potential was characterized after infection by Toxoplasma gondii. Human monocytes infested by T. gondii were stained with two membrane potential sensitive dyes, DiOC(6)(3) carbocyanine and DiSBAC(2)(3) bis-oxonol, before fluorescence emission analysis by confocal laser ... More
Diacylglycerol induces fusion of nuclear envelope membrane precursor vesicles.
'Purified membrane vesicles isolated from sea urchin eggs form nuclear envelopes around sperm nuclei following GTP hydrolysis in the presence of cytosol. A low density subfraction of these vesicles (MV1), highly enriched in phosphatidylinositol (PtdIns), is required for nuclear envelope formation. Membrane fusion of MV1 with a second fraction that ... More
Role of the cytoskeleton in calcium signaling in NIH 3T3 cells. An intact cytoskeleton is required for agonist-induced [Ca2+]i signaling, but not for capacitative calcium entry.
AuthorsRibeiro CM, Reece J, Putney JW
JournalJ Biol Chem
PubMed ID9334235
'Treatment of NIH 3T3 cells with cytochalasin D (10 microM, 1 h at 37 degrees C) disrupted the actin cytoskeleton and changed the cells from a planar, extended morphology, to a rounded shape. Calcium mobilization by ATP or by platelet-derived growth factor was abolished, while the ability of thapsigargin (2 ... More
Mercury-induced apoptosis in human lymphocytes: caspase activation is linked to redox status.
AuthorsShenker BJ, Pankoski L, Zekavat A, Shapiro IM
JournalAntioxid Redox Signal
PubMed ID12215206
'There is growing evidence that heavy metals, in general, and mercurial compounds, in particular, are toxic to the human immune system. We have previously shown that methyl mercuric chloride (MeHgCl) is a potent human T-cell apoptogen; moreover, mitochondria appear to be a target organelle for the induction of cell death. ... More
Epithelial intestinal cell apoptosis induced by Helicobacter pylori depends on expression of the cag pathogenicity island phenotype.
'Helicobacter pylori has been shown to induce chronic active gastritis and peptic ulcer and may contribute to the development of duodenal ulcer. Previous studies have shown that H. pylori mediates apoptosis of gastric epithelial cells via a Fas-dependent pathway. However, evidence for the induction of such a mechanism in intestinal ... More
Penetration of Toxoplasma gondii into host cells induces changes in the distribution of the mitochondria and the endoplasmic reticulum.
Authorsde Melo EJ, de Carvalho TU, de Souza W
JournalCell Struct Funct
PubMed ID1473161
'Fluorescence microscopy, using dyes which specifically label mitochondria, endoplasmic reticulum and the Golgi complex, and transmission electron microscopy, were used to analyze the changes which occur in the organization of these structures during interaction of Toxoplasma gondii with host cells. In uninfected cells the mitochondria are long filamentous structures which ... More
Characterization of kinectin, a kinesin-binding protein: primary sequence and N-terminal topogenic signal analysis.
AuthorsYu H, Nicchitta CV, Kumar J, Becker M, Toyoshima I, Sheetz MP
JournalMol Biol Cell
PubMed ID7787244
'Kinectin is a kinesin-binding protein (Toyoshima et al., 1992) that is required for kinesin-based motility (Kumar et al., 1995). A kinectin cDNA clone containing a 4.7-kilobase insert was isolated from an embryonic chick brain cDNA library by immunoscreening with a panel of monoclonal antibodies. The cDNA contained an open reading ... More
Mitochondrial fusion in yeast requires the transmembrane GTPase Fzo1p.
'Membrane fusion is required to establish the morphology and cellular distribution of the mitochondrial compartment. In Drosophila, mutations in the fuzzy onions (fzo) GTPase block a developmentally regulated mitochondrial fusion event during spermatogenesis. Here we report that the yeast orthologue of fuzzy onions, Fzo1p, plays a direct and conserved role ... More
Accessibility of cell surface thiols in human lymphocytes is altered by ionophores or OKT-3 antibody.
AuthorsBalázs M, Matkó J, Szöllösi J, Mátyus L, Fulwyler MJ, Damjanovich S
JournalBiochem Biophys Res Commun
PubMed ID3675722
'The accessibility of cell surface sulfhydryl groups in human peripheral lymphocytes was investigated with 5,5''-dithiobis-(2-nitrobenzoic acid) in the presence and absence of ionophore antibiotics and the monoclonal antibody, OKT-3. Only a few accessible protein thiols have been found on the cells as demonstrated by labeling with a fluorescent non-penetrating thiol-marker, ... More
Dynamic behavior of endoplasmic reticulum in living cells.
AuthorsLee C, Chen LB
JournalCell
PubMed ID3383243
'Endoplasmic reticulum (ER) was studied by fluorescence microscopy of living CV-1 cells treated with the fluorescent carbocyanine dye DiOC6(3). Using video recording and image processing techniques, several distinct forms of highly localized movements of ER were documented, categorized, and analyzed in terms of mechanism and structural implications. These include tubule ... More
Methods for the assessment of mitochondrial membrane permeabilization in apoptosis.
AuthorsGalluzzi L, Zamzami N, de La Motte Rouge T, Lemaire C, Brenner C, Kroemer G
JournalApoptosis
PubMed ID17294081
'Mitochondrial membrane permeabilization (MMP) is considered as the "point-of-no-return" in numerous models of programmed cell death. Indeed, mitochondria determine the intrinsic pathway of apoptosis, and play a major role in the extrinsic route as well. MMP affects the inner and outer mitochondrial membranes (IM and OM, respectively) to a variable ... More
Cyclosporin A depolarizes cytoplasmic membrane potential and interacts with Ca2+ ionophores.
AuthorsMátyus L, Balázs M, Aszalós A, Mulhern S, Damjanovich S
JournalBiochim Biophys Acta
PubMed ID3011116
'Cytoplasmic membrane potential of mouse lymphocytes was determined with flow cytometry and fluorescence spectroscopy using 3,3''-dihexylcarbocyanine iodide (DiOC6(3)). The amount of this lipophilic cation incorporated into the cytoplasmic membrane is dependent upon the transmembrane potential, so the dye is suitable for continuous monitoring of this parameter, under controlled conditions. Membrane ... More
A novel fission yeast gene, tht1+, is required for the fusion of nuclear envelopes during karyogamy.
AuthorsTange Y, Horio T, Shimanuki M, Ding DQ, Hiraoka Y, Niwa O
JournalJ Cell Biol
PubMed ID9442101
'We have isolated a fission yeast karyogamy mutant, tht1, in which nuclear congression and the association of two spindle pole bodies occurs but the subsequent fusion of nuclear envelopes is blocked. The tht1 mutation does not prevent meiosis, so cells execute meiosis with two unfused nuclei, leading to the production ... More
Membrane phosphatidylserine distribution as a non-apoptotic signalling mechanism in lymphocytes.
AuthorsElliott JI, Surprenant A, Marelli-Berg FM, Cooper JC, Cassady-Cain RL, Wooding C, Linton K, Alexander DR, Higgins CF
JournalNat Cell Biol
PubMed ID16025105
'Phosphatidylserine (PS) exposure is normally associated with apoptosis and the removal of dying cells. We observed that PS is exposed constitutively at high levels on T lymphocytes that express low levels of the transmembrane tyrosine phosphatase CD45RB. CD45 was shown to be a negative regulator of PS translocation in response ... More
Deoxyadenosine analogs induce programmed cell death in chronic lymphocytic leukemia cells by damaging the DNA and by directly affecting the mitochondria.
AuthorsGenini D, Adachi S, Chao Q, Rose DW, Carrera CJ, Cottam HB, Carson DA, Leoni LM
JournalBlood
PubMed ID11071652
'Adenine deoxynucleosides induce apoptosis in quiescent lymphocytes and are thus useful drugs for the treatment of indolent lymphoproliferative diseases. To explain why deoxyadenosine and its analogs are toxic to a cell that is not undergoing replicative DNA synthesis, several mechanisms have been proposed, including the direct binding of dATP to ... More
Stimulation of the pentose phosphate pathway and glutathione levels by dehydroascorbate, the oxidized form of vitamin C.
AuthorsPuskas F, Gergely P, Banki K, Perl A
JournalFASEB J
PubMed ID10877828
'Ascorbic acid, or vitamin C, generally functions as an antioxidant by directly reacting with reactive oxygen intermediates and has a vital role in defenses against oxidative stress. However, ascorbic acid also has pro-oxidant properties and may cause apoptosis of lymphoid and myeloid cells. The present study shows that dehydroascorbate, the ... More
Comparison of the effect of mitochondrial inhibitors on mitochondrial membrane potential in two different cell lines using flow cytometry and spectrofluorometry.
AuthorsKalbácová M, Vrbacký M, Drahota Z, Melková Z
JournalCytometry A
PubMed ID12655654
'BACKGROUND: Determination of mitochondrial membrane potential (DeltaPsim) is widely used to characterize cellular metabolism, viability, and apoptosis. Changes of DeltaPsim induced by inhibitors of oxidative phosphorylation characterize respective contributions of mitochondria and glycolysis to adenosine triphosphate (ATP) synthesis. METHODS: DeltaPsim in BSC-40 and HeLa G cell lines was determined by ... More
Integrin alpha IIb beta 3-dependent calcium signals regulate platelet-fibrinogen interactions under flow. Involvement of phospholipase C gamma 2.
AuthorsGoncalves I, Hughan SC, Schoenwaelder SM, Yap CL, Yuan Y, Jackson SP
JournalJ Biol Chem
PubMed ID12832405
'Platelet adhesion to fibrinogen is important for platelet aggregation and thrombus growth. In this study we have examined the mechanisms regulating platelet adhesion on immobilized fibrinogen under static and shear conditions. We demonstrate that integrin alpha IIb beta 3 engagement of immobilized fibrinogen is sufficient to induce an oscillatory calcium ... More
Mag-indo1 affinity for Ca(2+), compartmentalization and binding to proteins: the challenge of measuring Mg(2+) concentrations in living cells.
AuthorsPesco J, Salmon JM, Vigo J, Viallet P
JournalAnal Biochem
PubMed ID11237323
'A physicochemical study of the Mag-indo1 binding to Ca(2+) in solution showed that: (i) the characteristic fluorescence spectra of Ca(2+)-bound and Mg(2+)-bound Mag-indo1 are identical; (ii) two successive equilibria occur for increasing Ca(2+) concentrations; and (iii) the value of the dissociation constant of the first one, as determined by using ... More
Selective inhibition of dipeptidyl peptidase I, not caspases, prevents the partial processing of procaspase-3 in CD3-activated human CD8(+) T lymphocytes.
AuthorsBidère N, Briet M, Dürrbach A, Dumont C, Feldmann J, Charpentier B, de Saint-Basile G, Senik A
JournalJ Biol Chem
PubMed ID12080079
'Activation of primary human T cells by anti-CD3 and interleukin-2 resulted in partial processing of procaspase-3 in activated nonapoptotic (Delta Psi(m)high) CD8(+) T cells but not in CD4(+) T cells. Apical caspases-8 and -9 were not activated, and Bid was not processed to truncated Bid. Boc-D.fmk, a broad spectrum caspase ... More
Poly(ADP-ribose) synthetase activation mediates mitochondrial injury during oxidant-induced cell death.
AuthorsVirág L, Salzman AL, Szabó C
JournalJ Immunol
PubMed ID9759901
'Reactive oxidant species are important mediators of tissue injury in shock, inflammation, and reperfusion injury. The actions of a number of these oxidants (e.g., hydroxyl radical and peroxynitrite, a reactive oxidant produced by the reaction of nitric oxide and superoxide) are mediated in part by the activation of the nuclear ... More
Flow cytometric evaluation of apoptosis, necrosis and recovery when culturing monocytes.
'After developing and applying a method for cryopreserving monocytes, we found a substantial cell loss when culturing these cells. Monocytes were isolated from blood donors by density gradient centrifugation, purified by elutriation and cryopreserved. Thawed cells were cultured in ultra low attachment wells and studied with Annexin V, Propidium iodide, ... More
A role of the mitochondrial apoptosis-inducing factor in granulysin-induced apoptosis.
AuthorsPardo J, Pérez-Galán P, Gamen S, Marzo I, Monleón I, Kaspar AA, Susín SA, Kroemer G, Krensky AM, Naval J, Anel A
JournalJ Immunol
PubMed ID11466337
'Granulysin is a cytolytic molecule released by CTL via granule-mediated exocytosis. In a previous study we showed that granulysin induced apoptosis using both caspase- and ceramide-dependent and -independent pathways. In the present study we further characterize the biochemical mechanism for granulysin-induced apoptosis of tumor cells. Granulysin-induced death is significantly inhibited ... More
Association of cytoplasmic free Ca2+ gradients with subcellular organelles.
AuthorsWahl M, Sleight RG, Gruenstein E
JournalJ Cell Physiol
PubMed ID1537888
'Previous investigations have identified gradients of intracellular free (Ca2+)i (Ca2+i) in the cytoplasm of human fibroblasts. In this study we have compared the spatial distribution of these gradients with the subcellular distribution of cytoplasmic organelles. Using the Ca(2+)-sensitive dye fura-2 and organelle-specific fluorescent dyes, we have found that the highest ... More
TCR-mediated up-regulation of c-FLIPshort correlates with resistance toward CD95-mediated apoptosis by blocking death-inducing signaling complex activity.
AuthorsKirchhoff S, Müller WW, Krueger A, Schmitz I, Krammer PH
JournalJ Immunol
PubMed ID11086065
'To investigate apoptosis resistance upon restimulation in human peripheral blood T lymphocytes, we used the following in vitro model. This model represents the main features of T cell reactivity: freshly isolated PHA-activated T cells cultured in IL-2 for a prolonged period of time develop a CD95 (APO-1/Fas) apoptosis-sensitive phenotype. These ... More
Actomyosin-based motility of endoplasmic reticulum and chloroplasts in Vallisneria mesophyll cells.
AuthorsLiebe S, Menzel D
JournalBiol Cell
PubMed ID8785522
'Intracellular localization and motile behaviour of the endoplasmic reticulum (ER), plastids and mitochondria were studied in living mesophyll cells of Vallisneria using the vital fluorochrome 3,3'-dihexyloxacarbocyanine iodide (DIOC6(3)). In quiescent cells, the ER was composed of a three-dimensional network of tubular and lamellar elements. Chloroplasts were distributed evenly throughout the ... More
JC-1, but not DiOC6(3) or rhodamine 123, is a reliable fluorescent probe to assess delta psi changes in intact cells: implications for studies on mitochondrial functionality during apoptosis.
AuthorsSalvioli S, Ardizzoni A, Franceschi C, Cossarizza A
JournalFEBS Lett
PubMed ID9247146
'The sensitivity and specificity of three fluorescent probes used for cytofluorimetric analysis of mitochondrial membrane potential (delta psi) were studied in the U937 human cell line. First, the role of plasmamembrane in influencing the binding of the probes to mitochondria has been investigated. The depolarization of plasmamembrane with high doses ... More
Bioenergetics of the staphylococcal multidrug export protein QacA. Identification of distinct binding sites for monovalent and divalent cations.
AuthorsMitchell BA, Paulsen IT, Brown MH, Skurray RA
JournalJ Biol Chem
PubMed ID9920900
'The multidrug efflux pump QacA from Staphylococcus aureus confers resistance to an extensive range of structurally dissimilar compounds. Fluorimetric analyses demonstrated that QacA confers resistance to the divalent cation 4'',6-diamidino-2-phenylindole, utilizing a proton motive force-dependent efflux mechanism previously demonstrated for QacA-mediated resistance to the monovalent cation ethidium. Both the ionophores ... More
Bid-induced cytochrome c release is mediated by a pathway independent of mitochondrial permeability transition pore and Bax.
AuthorsKim TH, Zhao Y, Barber MJ, Kuharsky DK, Yin XM
JournalJ Biol Chem
PubMed ID10982793
'Bid, a pro-apoptosis "BH3-only" member of the Bcl-2 family, can be cleaved by caspase-8 after Fas/TNF-R1 engagement. The p15 form of truncated Bid (tBid) translocates to mitochondria and induces cytochrome c release, leading to the activation of downstream caspases and apoptosis. In the current study, we investigated the mechanism by ... More
Entry of poliovirus into cells is blocked by valinomycin and concanamycin A.
AuthorsIrurzun A, Carrasco L
JournalBiochemistry
PubMed ID11297425
'Poliovirus contains a virus particle devoid of a lipid envelope that does not require an intact pH to enter into susceptible cells. Thus, the blockade of pH gradient generated in endosomes is not sufficient to impede the translocation of poliovirus particles to the cytoplasm, suggesting that translocation takes place at ... More
A high-throughput assay for mitochondrial membrane potential in permeabilized yeast cells.
AuthorsFarrelly E, Amaral MC, Marshall L, Huang SG
JournalAnal Biochem
PubMed ID11399043
'A fluorometric assay for mitochondrial membrane potential in permeabilized yeast cells has been developed. This method involves permeabilizing the plasma membrane and measuring the distribution of a mitochondrial membrane potential sensitive probe 3,3''-dipropylthiadicarbocyanine iodide (DiSC(3)(5); DiSC(3)). In permeabilized cells, DiSC(3) fluorescence decreased when introduced into energized mitochondria and increased three- ... More
Dynamics of the endoplasmic reticulum and other membranous organelles in growth cones of cultured neurons.
AuthorsDailey ME, Bridgman PC
JournalJ Neurosci
PubMed ID2723756
'The fluorescent lipophilic dye 3,3''-dihexyloxacarbocyanine iodide [DiOC6(3)] was used to examine the distribution of membrane-bound organelles in growth cones of cultured rat sympathetic neurons. Within chemically fixed growth cones, intense DiOC6(3) fluorescence was localized predominately to the base or central region of growth cones. However, in most growth cones several ... More
Different subcellular localization of Saccharomyces cerevisiae HMG-CoA reductase isozymes at elevated levels corresponds to distinct endoplasmic reticulum membrane proliferations.
AuthorsKoning AJ, Roberts CJ, Wright RL
JournalMol Biol Cell
PubMed ID8744950
'In all eucaryotic cell types analyzed, proliferations of the endoplasmic reticulum (ER) can be induced by increasing the levels of certain integral ER proteins. One of the best characterized of these proteins is HMG-CoA reductase, which catalyzes the rate-limiting step in sterol biosynthesis. We have investigated the subcellular distributions of ... More
Incorporation of fluorescently labeled contractile proteins into freshly isolated living adult cardiac myocytes.
'When fluorescently labeled contractile proteins are injected into embryonic muscle cells, they become incorporated into the cells'' myofibrils. In order to determine if this exchange of proteins is unique to the embryonic stage of development, we isolated adult cardiac myocytes and microinjected them with fluorescently labeled actin, myosin light chains, ... More
Role of MMM1 in maintaining mitochondrial morphology in Neurospora crassa.
AuthorsProkisch H, Neupert W, Westermann B
JournalMol Biol Cell
PubMed ID10982393
'Mmm1p is a protein required for maintenance of mitochondrial morphology in budding yeast. It was proposed that it is required to mediate the interaction of the mitochondrial outer membrane with the actin cytoskeleton. We report the cloning and characterization of MMM1 of the filamentous fungus Neurospora crassa, an organism that ... More
Quantitative assay by flow cytometry of the mitochondrial membrane potential in intact cells.
AuthorsRottenberg H, Wu S
JournalBiochim Biophys Acta
PubMed ID9739168
'Mitochondrial membrane potential, in situ, is an important indicator of mitochondrial function and dysfunction. Because of recent interest in the role of mitochondria in signaling, cell injury and cell death, there is a need for a convenient, sensitive and accurate method for the measurement of the mitochondrial membrane potential, Deltapsim, ... More
Tumoricidal activity of monocyte-derived dendritic cells: evidence for a caspase-8-dependent, Fas-associated death domain-independent mechanism.
AuthorsVanderheyde N, Aksoy E, Amraoui Z, Vandenabeele P, Goldman M, Willems F
JournalJ Immunol
PubMed ID11564767
'Monocyte-derived dendritic cells (DC) were found to be cytotoxic for several tumor cell lines including Jurkat cells, which were killed through a calcium-independent pathway. K562 cells were resistant, excluding a NK cell-like activity. DC-mediated apoptosis did not involve classical death receptors because it was not reversed by blocking TNF/TNFR, CD95/CD95 ... More
Electrophysiology of T lymphocyte cholinergic receptors.
AuthorsShapiro HM, Strom TB
JournalProc Natl Acad Sci U S A
PubMed ID6968907
'The presence of functional muscarinic-cholinergic receptors on at least some T lymphocytes is suggested by the existence of saturable, high-affinity, specific muscarinic binding in T cell-enriched, but not in B cell-enriched, cell suspensions and by observed cholinergic effects on lymphocytes, (e.g., increased lytic capacity of cytotoxic lymphocytes preincubated with muscarinic ... More
Complex interplay between glutamate receptors and intracellular Ca2+ stores during ischaemia in rat spinal cord white matter.
AuthorsOuardouz M, Malek S, Coderre E, Stys PK
JournalJ Physiol
PubMed ID16945971
'Electrophysiological recordings of propagated compound action potentials (CAPs) and axonal Ca(2+) measurements using confocal microscopy were used to study the interplay between AMPA receptors and intracellullar Ca(2+) stores in rat spinal dorsal columns subjected to in vitro combined oxygen and glucose deprivation (OGD). Removal of Ca(2+) or Na(+) from the ... More
Loss of function of cytochrome c in Jurkat cells undergoing fas-mediated apoptosis.
AuthorsKrippner A, Matsuno-Yagi A, Gottlieb RA, Babior BM
JournalJ Biol Chem
PubMed ID8702951
'Mitochondrial function was examined in Jurkat cells undergoing Fas-mediated apoptosis. With succinate or ascorbate/tetramethylphenylenediamine as substrate, oxygen uptake by digitonin-permeabilized apoptotic mitochondria was greatly decreased as compared with control. Assessment of the function of the cytochrome c-cytochrome oxidase segment of the electron transport chain of apoptotic mitochondria showed that the ... More
Implication of mitochondrial hydrogen peroxide generation in ceramide-induced apoptosis.
AuthorsQuillet-Mary A, Jaffrézou JP, Mansat V, Bordier C, Naval J, Laurent G
JournalJ Biol Chem
PubMed ID9261153
'The key events implicated in ceramide-triggered apoptosis remain unknown. In this study we show that 25 microM C6-ceramide induced significant H2O2 production within 60 min, which increased up to 180 min in human myeloid leukemia U937 cells. Inactive analogue dihydro-C6-ceramide had no effect. Furthermore, no H2O2 production was observed in ... More
Inositol trisphosphate and cyclic ADP-ribose-mediated release of Ca2+ from single isolated pancreatic zymogen granules.
'In pancreatic acinar cells low (physiological) agonist concentrations evoke cytosolic Ca2+ spikes specifically in the apical secretory pole that contains a high density of secretory (zymogen) granules (ZGs). Inositol 1,4,5-trisphosphate (IP3) is believed to release Ca2+ from the endoplasmic reticulum, but we have now tested whether the Ca(2+)-releasing messengers IP3 ... More
Dynamics of the genome during early Xenopus laevis development: karyomeres as independent units of replication.
AuthorsLemaitre JM, Géraud G, Méchali M
JournalJ Cell Biol
PubMed ID9732278
'During Xenopus laevis early development, the genome is replicated in less than 15 min every 30 min. We show that during this period, DNA replication proceeds in an atypical manner. Chromosomes become surrounded by a nuclear membrane lamina forming micronuclei or karyomeres. This genomic organization permits that prereplication centers gather ... More
The effect of Chlamydia trachomatis infection on the host cell cytoskeleton and membrane compartments.
AuthorsCampbell S, Richmond SJ, Yates PS
JournalJ Gen Microbiol
PubMed ID2483409
'Human epithelial cells and the McCoy cell line were infected with Chlamydia trachomatis, serotype E. The organization of the cytoplasm was then studied with probes which stained cytoskeletal components and membrane compartments. The major actin-containing stress fibre bundles were not associated with inclusions due to the peri-basal and peri-apical location ... More
Endoplasmic reticulum and Golgi-like elements in Entamoeba.
AuthorsMazzuco A, Benchimol M, De Souza W
JournalMicron
PubMed ID9377074
'The cytoplasm of Entamoeba is characterized by the presence of a large number of vesicles of different size and shape. Previous electron microscopic studies have not clearly revealed the presence of the endoplasmic reticulum and the Golgi complex. In the present study two approaches were used aimed at the identification ... More
Fiber cell denucleation in the primate lens.
AuthorsBassnett S
JournalInvest Ophthalmol Vis Sci
PubMed ID9286256
'PURPOSE: To determine the morphologic and biochemical events preceding the breakdown of fiber cell nuclei in the primate lens. METHODS: Monkey lens slices were labeled with fluorescent probes and optically sectioned using a confocal microscope. The distribution of nuclear histones was visualized by immunofluorescence. DNA and cellular membranes were imaged ... More
Structural variability of the sub-surface cisternae in intact, isolated outer hair cells shown by fluorescent labelling of intracellular membranes and freeze-fracture.
AuthorsForge A, Zajic G, Li L, Nevill G, Schacht J
JournalHear Res
PubMed ID8432688
'The intracellular membrane systems in intact, isolated outer hair cells were visualised using the fluorescent membrane probe 3,3''-dihexyloxacarbocyanine iodide (DiOC6) and by freeze-fracture, and f-actin distribution was examined with rhodamine-phalloidin. DiOC6 stained the sub-surface cisternal membranes in the lateral wall and revealed a membrane system running in the centre of ... More
Laser confocal scanning microscopy of the surface membrane/T-tubular system and the sarcoplasmic reticulum in insect striated muscle stained with DilC18(3).
AuthorsBaumann O, Kitazawa T, Somlyo AP
JournalJ Struct Biol
PubMed ID2100146
'The structure of the surface membrane/transverse tubular (T-tubular) system and of the sarcoplasmic reticulum (SR) of the labial adductor muscle of the honey bee (Apis mellifera) was examined by laser confocal scanning microscopy, after staining with the fluorescent membrane probe DiIC18(3). The following components of the surface membrane/T-tubular system were ... More
Changes in the endoplasmic reticulum structure of Paramecium primaurelia in relation to different cellular physiological states.
AuthorsRamoino P, Diaspro A, Fato M, Beltrame F, Robello M
JournalJ Photochem Photobiol B
PubMed ID10739141
'The fluorochrome 3,3''-dihexyloxacarbocyanine iodide [DiOC6(3)], a vital dye utilized to stain the endoplasmic reticulum (ER) of animal and plant cells, has been used to visualize the ER-type structures of Paramecium primaurelia under confocal laser scanning microscopy (CLSM). The morphology of the ER has been studied in paramecia in different physiological ... More
DS28-6, a temperature-sensitive mutant of Chinese hamster ovary cells, expresses key phenotypic changes associated with brefeldin A treatment.
AuthorsZuber C, Roth J, Misteli T, Nakano A, Moremen K
JournalProc Natl Acad Sci U S A
PubMed ID1946407
'The temperature-sensitive Chinese hamster ovary (CHO) cell mutant DS28-6 has been previously shown to be pleiotropically defective in protein secretion. We have examined the mutant cells to determine the intracellular site of the block in secretion. By transmission electron microscopy a time-dependent disassembly of the Golgi apparatus was found under ... More
Caspase dependence of target cell damage induced by cytotoxic lymphocytes.
AuthorsSarin A, Haddad EK, Henkart PA
JournalJ Immunol
PubMed ID9743340
'Since the CTL secreted granule protease granzyme B can activate multiple target caspases, it has been proposed that this pathway is responsible for CTL-induced cytolysis of Fas-negative targets. However, target lysis via the granule exocytosis pathway is completely resistant to caspase inhibitors. To test the possibility that granzymes trigger a ... More
Model system for plant cell biology: GFP imaging in living onion epidermal cells.
AuthorsScott A, Wyatt S, Tsou PL, Robertson D, Allen NS
JournalBiotechniques
PubMed ID10376152
'The ability to visualize organelle localization and dynamics is very useful in studying cellular physiological events. Until recently, this has been accomplished using a variety of staining methods. However, staining can give inaccurate information due to nonspecific staining, diffusion of the stain or through toxic effects. The ability to target ... More
DiOC6(3): a useful dye for staining the endoplasmic reticulum.
'The present review discusses the fluorescent organelle probe, DiOC6(3), with reference to its structure, chemistry, availability, spectral properties, labeling procedures, vital staining characteristics, and major applications in cellular and molecular biology. The specificity of dye for endoplasmic reticulum is summarized. We examine the simplicity and advantages of the fluorescent dye ... More
A subpopulation of mitochondria prevents cytosolic calcium overload in endothelial cells after cold ischemia/reperfusion.
AuthorsAmberger A, Weiss H, Haller T, Köck G, Hermann M, Widschwendter M, Margreiter R
JournalTransplantation
PubMed ID11455264
'BACKGROUND: Calcium represents a key mediator of cold ischemia/reperfusion (CIR) injury presumably by affecting mitochondrial function. In this study, we investigated cellular and mitochondrial changes of calcium homeostasis in sublethally damaged human endothelial cells. METHODS: Changes in cellular and mitochondrial calcium concentrations were studied after cold ischemia in University of ... 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
Flow cytometric probes of early events in cell activation.