Hematopoietic potential of stem cells isolated from murine skeletal muscle.
AuthorsJackson KA,Mi T,Goodell MA
JournalProceedings of the National Academy of Sciences of the United States of America
PubMed ID10588731
We have discovered that cells derived from the skeletal muscle of adult mice contain a remarkable capacity for hematopoietic differentiation. Cells prepared from muscle by enzymatic digestion and 5-day in vitro culture were harvested, and 18 Ã 10(3) cells were introduced into each of six lethally irradiated recipients together with ... More
P-glycoprotein-mediated Hoechst 33342 transport out of the lipid bilayer.
AuthorsShapiro AB, Corder AB, Ling V
JournalEur J Biochem
PubMed ID9431998
High-level expression of P-glycoprotein, a 170-kDa mammalian plasma membrane ATPase, is the cause of an important and widespread form of cancer multidrug resistance. P-glycoprotein reduces cellular accumulation of an enormous variety of lipophilic compounds. The basis for this broad substrate specificity is not well understood. We explored this issue by ... More
Assembly of the nuclear pore: biochemically distinct steps revealed with NEM, GTP gamma S, and BAPTA.
AuthorsMacaulay C, Forbes DJ
JournalJ Cell Biol
PubMed ID8567730
A key event in nuclear formation is the assembly of functional nuclear pores. We have used a nuclear reconstitution system derived from Xenopus eggs to examine the process of nuclear pore assembly in vitro. With this system, we have identified three reagents which interfere with nuclear pore assembly, NEM, GTP ... More
High-speed photodamage cell selection using bromodeoxyuridine/Hoechst 33342 photosensitized cell killing.
AuthorsHerweijer H, Stokdijk W, Visser JW
JournalCytometry
PubMed ID2452055
One of the major drawbacks of droplet sorting in a flow cytometer is the relatively low sorting speed. Thus, we have developed an alternative, faster sorting technique: photodamage cell sorting. In a photodamage cell sorter all unwanted cells, as detected with the first, measuring laser, are killed with the second, ... More
Cyclin G2 associates with protein phosphatase 2A catalytic and regulatory B' subunits in active complexes and induces nuclear aberrations and a G1/S phase cell cycle arrest.
AuthorsBennin David A; Don Aruni S Arachchige; Brake Tiffany; McKenzie Jennifer L; Rosenbaum Heidi; Ortiz Linette; DePaoli-Roach Anna A; Horne Mary C;
JournalJ Biol Chem
PubMed ID11956189
Cyclin G2, together with cyclin G1 and cyclin I, defines a novel cyclin family expressed in terminally differentiated tissues including brain and muscle. Cyclin G2 expression is up-regulated as cells undergo cell cycle arrest or apoptosis in response to inhibitory stimuli independent of p53 (Horne, M., Donaldson, K., Goolsby, G., ... More
Integration of NPY, AGRP, and melanocortin signals in the hypothalamic paraventricular nucleus: evidence of a cellular basis for the adipostat.
AuthorsCowley MA, Pronchuk N, Fan W, Dinulescu DM, Colmers WF, Cone RD
JournalNeuron
PubMed ID10677034
'Energy stores are held relatively constant in many mammals. The circuitry necessary for maintaining energy homeostasis should (1) sense the amount of energy stored in adipose tissue, (2) sense and integrate the multiple opposing signals regarding nutritional state, and (3) provide output regulating energy intake and expenditure to maintain energy ... More
Spatially organised mitochondrial calcium uptake through a novel pathway in chick neurones.
AuthorsCoatesworth W, Bolsover S
JournalCell Calcium
PubMed ID16338004
'A brief depolarisation of chick sensory neurones evokes a calcium increase in mitochondria that peaks 1-2s after the depolarisation event and then decays over tens of seconds. Peripheral mitochondria take up more calcium than do central ones, even when the cytosolic calcium increase is spatially homogeneous. The calcium influx into ... More
Caspase activation contributes to delayed death of heat-stressed striatal neurons.
AuthorsWhite MG, Emery M, Nonner D, Barrett JN
JournalJ Neurochem
PubMed ID14622126
'Hyperthermia can contribute to brain damage both during development and post-natally. We used rat embryonic striatal neurons in culture to study mechanisms underlying hyperthermia-induced neuronal death. Heat stress at 43 degrees C for 2 h produced no obvious signs of damage during the first 12 h after the stress, but ... More
Correlated light and electron microscopic imaging of multiple endogenous proteins using Quantum dots.
AuthorsGiepmans BN, Deerinck TJ, Smarr BL, Jones YZ, Ellisman MH
JournalNat Methods
PubMed ID16179920
'The importance of locating proteins in their context within cells has been heightened recently by the accomplishments in molecular structure and systems biology. Although light microscopy (LM) has been extensively used for mapping protein localization, many studies require the additional resolution of the electron microscope. Here we report the application ... More
Profiling antibody responses by multiparametric analysis of primary B cells.
AuthorsStory CM, Papa E, Hu CC, Ronan JL, Herlihy K, Ploegh HL, Love JC,
JournalProc Natl Acad Sci U S A
PubMed ID19004776
'Determining the efficacy of a vaccine generally relies on measuring neutralizing antibodies in sera. This measure cannot elucidate the mechanisms responsible for the development of immunological memory at the cellular level, however. Quantitative profiles that detail the cellular origin, extent, and diversity of the humoral (antibody-based) immune response would improve ... More
High-throughput discovery of synthetic surfaces that support proliferation of pluripotent cells.
'Synthetic materials that promote the growth or differentiation of cells have advanced the fields of tissue engineering and regenerative medicine. Most functional biomaterials are based on a handful of peptide sequences derived from protein ligands for cell surface receptors. Because few proteins possess short peptide sequences that alone can engage ... 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
A high-throughput, cell-based screening method for siRNA and small molecule inhibitors of mTORC1 signaling using the In Cell Western technique.
AuthorsHoffman GR, Moerke NJ, Hsia M, Shamu CE, Blenis J,
JournalAssay Drug Dev Technol
PubMed ID20085456
'The mTORC1 pathway is a central regulator of cell growth, and defective mTORC1 regulation plays a causative role in a variety of human diseases, including cancer, tumor syndromes such as the tuberous sclerosis complex (TSC) and lymphangioleiomyomatosis (LAM), and metabolic diseases such as diabetes and obesity. Given the importance of ... More
Titrating the effects of mitochondrial complex I impairment in the cell physiology.
AuthorsBarrientos A, Moraes CT
JournalJ Biol Chem
PubMed ID10347173
'The mitochondrial oxidative phosphorylation system consists of five multimeric enzymes (complexes I-V). NADH dehydrogenase or complex I (CI) is affected in most of the mitochondrial diseases and in some neurodegenerative disorders. We have studied the physiological consequences of a partial CI inhibition at the cellular level. We used a genetic ... More
Identification of a subpopulation of cells with cancer stem cell properties in head and neck squamous cell carcinoma.
AuthorsPrince ME, Sivanandan R, Kaczorowski A, Wolf GT, Kaplan MJ, Dalerba P, Weissman IL, Clarke MF, Ailles LE
JournalProc Natl Acad Sci U S A
PubMed ID17210912
'Like many epithelial tumors, head and neck squamous cell carcinoma (HNSCC) contains a heterogeneous population of cancer cells. We developed an immunodeficient mouse model to test the tumorigenic potential of different populations of cancer cells derived from primary, unmanipulated human HNSCC samples. We show that a minority population of CD44(+) ... More
Age-related macular degeneration. The lipofusion component N-retinyl-N-retinylidene ethanolamine detaches proapoptotic proteins from mitochondria and induces apoptosis in mammalian retinal pigment epithelial cells.
AuthorsSuter M, Remé C, Grimm C, Wenzel A, Jäättela M, Esser P, Kociok N, Leist M, Richter C
JournalJ Biol Chem
PubMed ID11006290
'10-20% of individuals over the age of 65 suffer from age-related macular degeneration (AMD), the leading cause of severe visual impairment in humans living in developed countries. The pathogenesis of this complex disease is poorly understood, and no efficient therapy or prevention exists to date. A precondition for AMD appears ... More
PMN transendothelial migration decreases nuclear NFkappaB in IL-1beta-activated endothelial cells: role of PECAM-1.
'During the systemic inflammatory response, circulating cytokines interact with the vascular endothelium, resulting in activation and nuclear accumulation of the nuclear transcription factor, nuclear factor kappa B (NFkappaB). In turn, NFkappaB transactivates relevant proinflammatory genes, resulting in an amplification of the inflammatory response. Because this scenario is potentially detrimental to ... More
Quantum dot-induced epigenetic and genotoxic changes in human breast cancer cells.
AuthorsChoi AO, Brown SE, Szyf M, Maysinger D,
JournalJ Mol Med
PubMed ID17965848
'The staggering array of nanotechnological products, found in our environment and those applicable in medicine, has stimulated a growing interest in examining their long-term impact on genetic and epigenetic processes. We examined here the epigenomic and genotoxic response to cadmium telluride quantum dots (QDs) in human breast carcinoma cells. QD ... More
Signaling through extracellular signal-regulated kinase is required for spermatogonial proliferative response to stem cell factor.
AuthorsDolci S, Pellegrini M, Di Agostino S, Geremia R, Rossi P
JournalJ Biol Chem
PubMed ID11502745
'In vitro addition of stem cell factor (SCF) to c-kit-expressing A(1)-A(4) spermatogonia from prepuberal mice stimulates their progression into the mitotic cell cycle and significantly reduces apoptosis in these cells. SCF addition results in a transient activation of extracellular signal-regulated kinases (Erk)1/2 as well as of phosphatidylinositol 3-kinase (PI3K)-dependent Akt ... More
The small conductance K+ channel, KCNQ1: expression, function, and subunit composition in murine trachea.
AuthorsGrahammer F, Warth R, Barhanin J, Bleich M, Hug MJ
JournalJ Biol Chem
PubMed ID11527966
'The gene KCNQ1 encodes a K(+) channel alpha-subunit important for cardiac repolarization, formerly known as K(v)LQT1. In large and small intestine a channel complex consisting of KCNQ1 and the beta-subunit KCNE3 (MiRP2) is known to mediate the cAMP-activated basolateral K(+) current, which is essential for luminal Cl(-) secretion. Northern blot ... More
A role for the extracellular signal-regulated kinase and p38 mitogen-activated protein kinases in interleukin-1 beta-stimulated delayed signal tranducer and activator of transcription 3 activation, atrial natriuretic factor expression, and cardiac myocyte morphology.
AuthorsNg DC, Long CS, Bogoyevitch MA
JournalJ Biol Chem
PubMed ID11382751
'We have demonstrated that two hypertrophic agents, interleukin-1 beta (IL-1 beta) and leukemic inhibitory factor (LIF), altered cardiac myocyte morphology with striking similarity and prompted us to investigate the common actions of these cytokines. We compared the phosphorylation/activation of signal tranducer and activator of transcription 3 (STAT3), extracellular signal-regulated kinase ... More
The NEDD8 system is essential for cell cycle progression and morphogenetic pathway in mice.
AuthorsTateishi K, Omata M, Tanaka K, Chiba T
JournalJ Cell Biol
PubMed ID11696557
'NEDD8/Rub1 is a ubiquitin (Ub)-like molecule that covalently ligates to target proteins through an enzymatic cascade analogous to ubiquitylation. This modifier is known to target all cullin (Cul) family proteins. The latter are essential components of Skp1/Cul-1/F-box protein (SCF)-like Ub ligase complexes, which play critical roles in Ub-mediated proteolysis. To ... More
Modulation of acto-myosin contractility in skeletal muscle myoblasts uncouples growth arrest from differentiation.
AuthorsDhawan J, Helfman DM
JournalJ Cell Sci
PubMed ID15252113
'Cell-substratum interactions trigger key signaling pathways that modulate growth control and tissue-specific gene expression. We have previously shown that abolishing adhesive interactions by suspension culture results in G(0) arrest of myoblasts. We report that blocking intracellular transmission of adhesion-dependent signals in adherent cells mimics the absence of adhesive contacts. We ... More
Foxo3 is essential for the regulation of ataxia telangiectasia mutated and oxidative stress-mediated homeostasis of hematopoietic stem cells.
AuthorsYalcin S, Zhang X, Luciano JP, Mungamuri SK, Marinkovic D, Vercherat C, Sarkar A, Grisotto M, Taneja R, Ghaffari S,
JournalJ Biol Chem
PubMed ID18424439
'Unchecked accumulation of reactive oxygen species (ROS) compromises maintenance of hematopoietic stem cells. Regulation of ROS by the tumor suppressor protein ataxia telangiectasia mutated (ATM) is critical for preserving the hematopoietic stem cell pool. In this study we demonstrate that the Foxo3 member of the Forkhead Box O (FoxO) family ... More
Essential role of voltage-dependent anion channel in various forms of apoptosis in mammalian cells.
AuthorsShimizu S, Matsuoka Y, Shinohara Y, Yoneda Y, Tsujimoto Y
JournalJ Cell Biol
PubMed ID11266442
'Through direct interaction with the voltage-dependent anion channel (VDAC), proapoptotic members of the Bcl-2 family such as Bax and Bak induce apoptogenic cytochrome c release in isolated mitochondria, whereas BH3-only proteins such as Bid and Bik do not directly target the VDAC to induce cytochrome c release. To investigate the ... More
Restriction of receptor movement alters cellular response: physical force sensing by EphA2.
AuthorsSalaita K, Nair PM, Petit RS, Neve RM, Das D, Gray JW, Groves JT,
JournalScience
PubMed ID20223987
'Activation of the EphA2 receptor tyrosine kinase by ephrin-A1 ligands presented on apposed cell surfaces plays important roles in development and exhibits poorly understood functional alterations in cancer. We reconstituted this intermembrane signaling geometry between live EphA2-expressing human breast cancer cells and supported membranes displaying laterally mobile ephrin-A1. Receptor-ligand binding, ... More
Binding to cadherins antagonizes the signaling activity of beta-catenin during axis formation in Xenopus.
AuthorsFagotto F, Funayama N, Gluck U, Gumbiner BM
JournalJ Cell Biol
PubMed ID8601588
'beta-Catenin, a cytoplasmic protein known for its association with cadherin cell adhesion molecules, is also part of a signaling cascade involved in embryonic patterning processes such as the determination of the dorsoventral axis in Xenopus and determination of segment polarity in Drosophila. Previous studies suggest that increased cytoplasmic levels of ... 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
Transplantation of quail collagen-tailed acetylcholinesterase molecules onto the frog neuromuscular synapse.
AuthorsRotundo RL, Rossi SG, Anglister L
JournalJ Cell Biol
PubMed ID9015307
'The highly organized pattern of acetylcholinesterase (AChE) molecules attached to the basal lamina of the neuromuscular junction (NMJ) suggests the existence of specific binding sites for their precise localization. To test this hypothesis we immunoaffinity purified quail globular and collagen-tailed AChE forms and determined their ability to attach to frog ... More
Interaction of Hsp90 with the nascent form of the mutant epidermal growth factor receptor EGFRvIII.
AuthorsLavictoire SJ, Parolin DA, Klimowicz AC, Kelly JF, Lorimer IA
JournalJ Biol Chem
PubMed ID12471035
'EGFRvIII is a mutant epidermal growth factor that promotes aggressive growth of glioblastomas. We made a plasmid that directed the expression of an EGFRvIII with three copies of the Flag epitope at its amino terminus. Flag-tagged EGFRvIII was expressed at the same levels as unmodified EGFRvIII, and showed the same ... More
Studies on the phenotype of migrant thymic stem cells.
AuthorsSuniara RK, Jenkinson EJ, Owen JJ
JournalEur J Immunol
PubMed ID9933088
'Stem cells first enter the thymus around the 11th to 12th days of gestation in BALB/c mouse embryos. The phenotype of these stem cells has been difficult to determine because their entry occurs when the thymic primordium is very small and involves too few stem cells to allow studies by ... More
Cell type specific applicability of 5-ethynyl-2'-deoxyuridine (EdU) for dynamic proliferation assessment in flow cytometry.
AuthorsDiermeier-Daucher S, Clarke ST, Hill D, Vollmann-Zwerenz A, Bradford JA, Brockhoff G,
JournalCytometry A
PubMed ID19235202
'Using the nucleoside analogue EdU (5-ethynyl-2''-deoxyuridine) for thymidine substitution instead of BrdU (5-bromo-2''-deoxyuridine) in cell proliferation assays has recently been proposed. However, the effect of EdU on cell viability, DNA synthesis, and cell cycle progression and consequently its usability for dynamic cell proliferation analysis in vitro has not been explored. ... More
SM-20 is a novel mitochondrial protein that causes caspase-dependent cell death in nerve growth factor-dependent neurons.
AuthorsLipscomb EA, Sarmiere PD, Freeman RS
JournalJ Biol Chem
PubMed ID11060309
'Sympathetic neurons undergo protein synthesis-dependent apoptosis when deprived of nerve growth factor (NGF). Expression of SM-20 is up-regulated in NGF-deprived sympathetic neurons, and ectopic SM-20 is sufficient to promote neuronal death in the presence of NGF. We now report that SM-20 is a mitochondrial protein that promotes cell death through ... More
Rootletin, a novel coiled-coil protein, is a structural component of the ciliary rootlet.
AuthorsYang J, Liu X, Yue G, Adamian M, Bulgakov O, Li T
JournalJ Cell Biol
PubMed ID12427867
'The ciliary rootlet, first recognized over a century ago, is a prominent structure originating from the basal body at the proximal end of a cilium. Despite being the largest cytoskeleton, its structural composition has remained unknown. Here, we report a novel 220-kD protein, designated rootletin, found in the rootlets of ... More
Cell damage-induced conformational changes of the pro-apoptotic protein Bak in vivo precede the onset of apoptosis.
AuthorsGriffiths GJ, Dubrez L, Morgan CP, Jones NA, Whitehouse J, Corfe BM, Dive C, Hickman JA
JournalJ Cell Biol
PubMed ID10085290
'Investigation of events committing cells to death revealed that a concealed NH2-terminal epitope of the pro-apoptotic protein Bak became exposed in vivo before apoptosis. This occurred after treatment of human Jurkat or CEM-C7A T-lymphoma cells with the mechanistically disparate agents staurosporine, etoposide or dexamethasone. The rapid, up to 10-fold increase ... More
Striated muscle preferentially expressed genes alpha and beta are two serine/threonine protein kinases derived from the same gene as the aortic preferentially expressed gene-1.
AuthorsHsieh CM, Fukumoto S, Layne MD, Maemura K, Charles H, Patel A, Perrella MA, Lee ME
JournalJ Biol Chem
PubMed ID10973969
'Aortic preferentially expressed gene (APEG)-1 is a 1.4-kilobase pair (kb) mRNA expressed in vascular smooth muscle cells and is down-regulated by vascular injury. An APEG-1 5''-end cDNA probe identified three additional isoforms. The 9-kb striated preferentially expressed gene (SPEG)alpha and the 11-kb SPEGbeta were found in skeletal muscle and heart. ... More
Polyglutamine-expanded ataxin-7 antagonizes CRX function and induces cone-rod dystrophy in a mouse model of SCA7.
AuthorsLa Spada AR, Fu YH, Sopher BL, Libby RT, Wang X, Li LY, Einum DD, Huang J, Possin DE, Smith AC, Martinez RA, Koszdin KL, Treuting PM, Ware CB, Hurley JB, Ptácek LJ, Chen S
JournalNeuron
PubMed ID11580893
'Spinocerebellar ataxia type 7 (SCA7) is an autosomal dominant disorder caused by a CAG repeat expansion. To determine the mechanism of neurotoxicity, we produced transgenic mice and observed a cone-rod dystrophy. Nuclear inclusions were present, suggesting that the disease pathway involves the nucleus. When yeast two-hybrid assays indicated that cone-rod ... More
Two-photon fluorescence absorption and emission spectra of dyes relevant for cell imaging.
AuthorsBestvater F, Spiess E, Stobrawa G, Hacker M, Feurer T, Porwol T, Berchner-Pfannschmidt U, Wotzlaw C, Acker H
JournalJ Microsc
PubMed ID12423261
'Two-photon absorption and emission spectra for fluorophores relevant in cell imaging were measured using a 45 fs Ti:sapphire laser, a continuously tuneable optical parametric amplifier for the excitation range 580-1150 nm and an optical multichannel analyser. The measurements included DNA stains, fluorescent dyes coupled to antibodies as well as organelle ... More
Unreliability of the cytochrome c-enhanced green fluorescent fusion protein as a marker of cytochrome c release in cells that overexpress Bcl-2.
AuthorsUnkila M, McColl KS, Thomenius MJ, Heiskanen K, Distelhorst CW
JournalJ Biol Chem
PubMed ID11489892
'A cytochrome c-enhanced green fluorescent protein chimera (cyt-c.EGFP) was used to monitor the release of cytochrome c from mitochondria in Bcl-2-negative and Bcl-2-positive MDA-MB-468 breast cancer cells. A comparison was made with the intracellular distribution of endogenous cytochrome c based on Western blotting of cell fractions and immunocytochemistry. The release ... More
Improved immunocytochemical identification of neural, endothelial, and inflammatory cell types in paraffin-embedded injured adult rat spinal cord.
AuthorsCasella GT, Bunge MB, Wood PM
JournalJ Neurosci Methods
PubMed ID15351516
'Methods that facilitate the accurate counting of specific neural cell types would be of substantial value in evaluating the efficacy of treatments applied to spinal cord injury. This report describes reliable procedures for identification of neurons, oligodendrocytes, astrocytes, endothelial cells and inflammatory cells (neutrophils and activated macrophage/microglial cells) in paraformaldehyde-fixed, ... More
Retinoic acid evoked-differentiation of neuroblastoma cells predominates over growth factor stimulation: an automated image capture and quantitation approach to neuritogenesis.
AuthorsSimpson PB, Bacha JI, Palfreyman EL, Woollacott AJ, McKernan RM, Kerby J
JournalAnal Biochem
PubMed ID11757502
'To facilitate the characterization of compounds that have positive growth factor mimetic effects on neuritogenesis, we have implemented a high-throughput functional assay which measures, in a multiparametric manner, the proliferation and differentiation characteristics of cells in a microtiter plate. Conditions were established using chronic incubation of SH-SY5Y human neuroblastoma cells ... More
Effect of mutant alpha-synuclein on dopamine homeostasis in a new human mesencephalic cell line.
'Mutations in alpha-synuclein have been linked to rare, autosomal dominant forms of Parkinson''s disease. Despite its ubiquitous expression, mutant alpha-synuclein primarily leads to the loss of dopamine-producing neurons in the substantia nigra. alpha-Synuclein is a presynaptic nerve terminal protein of unknown function, although several studies suggest it is important for ... More
Analysis of P-glycoprotein-mediated membrane transport in human peripheral blood lymphocytes using the UIC2 shift assay.
AuthorsPark SW, Lomri N, Simeoni LA, Fruehauf JP, Mechetner E
JournalCytometry A
PubMed ID12766968
'BACKGROUND: During transport-associated adenosine triphosphate hydrolysis, P-glycoprotein (Pgp) undergoes conformation transitions detected by UIC2, a functional anti-Pgp monoclonal antibody. A newly developed UIC2 shift assay is based on increased UIC2 reactivity in the presence of Pgp substrates. All peripheral blood leukocytes express low Pgp levels. The existing antibody-based detection methods ... More
Necrosis and apoptosis of polymorphonuclear cells exposed to peritoneal dialysis fluids in vitro.
'Conventional peritoneal dialysis (PD) fluids are known to inhibit polymorphonuclear cells (PMN) phagocytosis, oxidative burst and enzyme release. However, the relative contributions of apoptosis and/or necrosis to this dysfunction have not been examined. We investigated the effects of osmolality, glucose concentration and heat-sterilization of PD fluids on necrosis and apoptosis ... More
Defects in the ubiquitin pathway induce caspase-independent apoptosis blocked by Bcl-2.
AuthorsMonney L, Otter I, Olivier R, Ozer HL, Haas AL, Omura S, Borner C
JournalJ Biol Chem
PubMed ID9497330
'Apoptosis requires the activation of caspases (formerly interleukin 1beta-converting enzyme-like proteases), in particular those related to the caspase-3/7/6 subfamily. Recent data, however, revealed that, although caspase-specific inhibitors delay apoptosis, they are often incapable of preventing it. To obtain evidence for caspase-independent steps of apoptosis, we artificially created a high amount ... More
Modulation of vincristine sensitivity of human kidney tumor cells by pharmacological agents interfering with intracellular signals. No apparent relationship to changes in cytoplasmic Ca2+ or pH.
AuthorsNygren P, Larsson R
JournalBiochim Biophys Acta
PubMed ID2191723
'The effect of substances proposed to modulate intracellular signal systems on growth and sensitivity to vincristine in the human kidney tumor cell line ACHN was investigated and related to changes in cytoplasmic free Ca2+ concentration ([Ca2+]i) and cytoplasmic pH (pHi). Presence during culture of the protein kinase C (PKC) activator ... More
Induction of terminal differentiation in epithelial cells requires polymerization of hensin by galectin 3.
'During terminal differentiation, epithelia become columnar and develop specialized apical membrane structures (microvilli) and functions (regulated endocytosis and exocytosis). Using a clonal intercalated epithelial cell line, we found that high seeding density induced these characteristics, whereas low density seeding maintained a protoepithelial state. When cells were plated at low density, ... More
Receptor binding of NBD-labeled fluorescent estrogens and progestins in whole cells and cell-free preparations.
AuthorsCarlson KE, Coppey M, Magdelenat H, Katzenellenbogen JA
JournalJ Steroid Biochem
PubMed ID2539536
'We have studied the interactions of four fluorescent steroid conjugates with either the estrogen or progesterone receptor, both in whole cells and cell-free receptor preparations. The fluorophore, nitrobenzoxadiazole (NBD), was conjugated with a synthetic progestin, with a steroidal estrogen, a non-steroidal estrogen, and with an antiestrogen. With all compounds, receptor-specific ... More
Analyses of DNA content of living spermatozoa using flow cytometry techniques.
AuthorsDresser DW, Atkins CJ, Pinder A, Morrell JM
JournalJ Reprod Fertil
PubMed ID7692043
'A two-dimensional fluorescence analysis of spermatozoa stained with Hoechst 33342 was carried out using an Epics V flow cytometer. This analysis involved the measurement of fluorescence both in the conventional manner at 90 degrees and at a narrow forward angle (nominally 0 degree) to the direction of the interrogating laser ... More
An epithelial cell destined for apoptosis signals its neighbors to extrude it by an actin- and myosin-dependent mechanism.
AuthorsRosenblatt J, Raff MC, Cramer LP
JournalCurr Biol
PubMed ID11728307
'BACKGROUND: Simple epithelia encase developing embryos and organs. Although these epithelia consist of only one or two layers of cells, they must provide tight barriers for the tissues that they envelop. Apoptosis occurring within these simple epithelia could compromise this barrier. How, then, does an epithelium remove apoptotic cells without ... More
Comparison of Hoechst 33342 and propidium iodide as fluorescent markers for sperm fusion with hamster oocytes.
AuthorsGreen DP
JournalJ Reprod Fertil
PubMed ID1285064
'Hamster oocytes were loaded with the DNA dyes Hoechst 33342 or propidium iodide. Oocytes incubated in 10 mumol Hoechst 333421(-1) showed intracellular fluorescence within 10-20 s of exposure, as did hamster and guinea-pig spermatozoa. Impaled oocytes to which acrosome-intact hamster spermatozoa were bound before injection of Hoechst 33342 showed dye ... More
The influenza A virus PB1-F2 protein targets the inner mitochondrial membrane via a predicted basic amphipathic helix that disrupts mitochondrial function.
'The 11th influenza A virus gene product is an 87-amino-acid protein provisionally named PB1-F2 (because it is encoded by an open reading frame overlapping the PB1 open reading frame). A significant fraction of PB1-F2 localizes to the inner mitochondrial membrane in influenza A virus-infected cells. PB1-F2 appears to enhance virus-induced ... More
Amyloid beta-peptide induces cell monolayer albumin permeability, impairs glucose transport, and induces apoptosis in vascular endothelial cells.
AuthorsBlanc EM, Toborek M, Mark RJ, Hennig B, Mattson MP
JournalJ Neurochem
PubMed ID9109512
'Amyloid beta-peptide (A beta) is deposited as insoluble fibrils in the brain parenchyma and cerebral blood vessels in Alzheimer''s disease (AD). In addition to neuronal degeneration, cerebral vascular alterations indicative of damage to vascular endothelial cells and disruption of the blood-brain barrier occur in AD. Here we report that A ... 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
Flow cytometric analysis of mammalian glial cultures treated with methotrexate.
AuthorsSerrano EE, Schimke RT
JournalGlia
PubMed ID2148556
'Methotrexate (MTX) is an antineoplastic drug that acts by competitive inhibition of the enzyme dihydrofolate reductase (DHFR). MTX treatment of cultured cell lines leads to the emergence of resistant cell populations. Studies using stepwise selection procedures have demonstrated that MTX resistance conferred by overproduction of DHFR can be caused by ... More
Guanidinylated neomycin delivers large, bioactive cargo into cells through a heparan sulfate-dependent pathway.
AuthorsElson-Schwab L, Garner OB, Schuksz M, Crawford BE, Esko JD, Tor Y
JournalJ Biol Chem
PubMed ID17311923
'Facilitating the uptake of molecules into living cells is of substantial interest for basic research and drug delivery applications. Arginine-rich peptides have been shown to facilitate uptake of high molecular mass cargos into cells, but the mechanism of uptake is complex and may involve multiple receptors. In this report, we ... More
Equilibrium binding of Hoechst 33258 and Hoechst 33342 fluorochromes with rat colorectal cells.
'We examined the biophysical characteristics of the interaction of Hoechst 33258 and 33342 dyes with normal rat colorectal cells as functions of fixation and solution composition. Classical dye-binding techniques were used to investigate the stoichiometry and binding constants with whole cells, and quantitative fluorescence image analysis was used to specifically ... More
Nuclear membrane dynamics and reassembly in living cells: targeting of an inner nuclear membrane protein in interphase and mitosis.
AuthorsEllenberg J, Siggia ED, Moreira JE, Smith CL, Presley JF, Worman HJ, Lippincott-Schwartz J
JournalJ Cell Biol
PubMed ID9298976
'The mechanisms of localization and retention of membrane proteins in the inner nuclear membrane and the fate of this membrane system during mitosis were studied in living cells using the inner nuclear membrane protein, lamin B receptor, fused to green fluorescent protein (LBR-GFP). Photobleaching techniques revealed the majority of LBR-GFP ... More
Developmental changes in the distribution of the endoplasmic reticulum and inositol 1,4,5-trisphosphate receptors and the spatial pattern of Ca2+ release during maturation of hamster oocytes.
AuthorsShiraishi K, Okada A, Shirakawa H, Nakanishi S, Mikoshiba K, Miyazaki S
JournalDev Biol
PubMed ID7649386
'During maturation of hamster oocytes, the distribution of the endoplasmic reticulum (ER) and inositol 1,4,5-trisphosphate receptors (InsP3Rs) was found to change dramatically, as observed using confocal microscopy with DiI and electron microscopy for the ER and immunohistochemistry for InsP3Rs. In immature oocytes at the germinal vesicle (GV) stage, ER and ... More
Diazoxide triggers cardioprotection against apoptosis induced by oxidative stress.
AuthorsIchinose M, Yonemochi H, Sato T, Saikawa T
JournalAm J Physiol Heart Circ Physiol
PubMed ID12623783
'Although mitochondrial ATP-sensitive potassium (mitoK(ATP)) channels have been reported to reduce the extent of apoptosis, the critical timing of mitoK(ATP) channel opening required to protect myocytes against apoptosis remains unclear. In the present study, we examined whether the mitoK(ATP) channel serves as a trigger of cardioprotection against apoptosis induced by ... More
In vivo migration of dendritic cells differentiated in vitro: a chimpanzee model.
AuthorsBarratt-Boyes SM, Watkins SC, Finn OJ
JournalJ Immunol
PubMed ID9144465
'Dendritic cells with potent Ag-presenting function can be propagated from peripheral blood using recombinant cytokines, and these cells have potential usefulness as immunotherapeutic agents in the treatment of cancer and other disease states. However, it is not known if these in vitro differentiated dendritic cells have the capacity to migrate ... More
Role of poly(ADP-ribose) polymerase (PARP) cleavage in apoptosis. Caspase 3-resistant PARP mutant increases rates of apoptosis in transfected cells.
AuthorsBoulares AH, Yakovlev AG, Ivanova V, Stoica BA, Wang G, Iyer S, Smulson M
JournalJ Biol Chem
PubMed ID10438458
'An early transient burst of poly(ADP-ribosyl)ation of nuclear proteins was recently shown to be required for apoptosis to proceed in various cell lines (Simbulan-Rosenthal, C., Rosenthal, D., Iyer, S., Boulares, H., and Smulson, M. (1998) J. Biol. Chem. 273, 13703-13712) followed by cleavage of poly(ADP-ribose) polymerase (PARP), catalyzed by caspase-3. ... More
Ionizing radiation down-regulates p53 protein in primary Egr-1-/- mouse embryonic fibroblast cells causing enhanced resistance to apoptosis.
AuthorsDas A, Chendil D, Dey S, Mohiuddin M, Mohiuddin M, Milbrandt J, Rangnekar VM, Ahmed MM
JournalJ Biol Chem
PubMed ID11035041
'In this study, we sought to investigate the mechanism of the proapoptotic function of Egr-1 in relation to p53 status in normal isogenic cell backgrounds by using primary MEF cells established from homozygous (Egr-1(-/-)) and heterozygous (Egr-1(+/-)) Egr-1 knock-out mice. Ionizing radiation caused significantly enhanced apoptosis in Egr-1(+/-) cells (22.8%; ... More
Differential effects of deuterium oxide on the fluorescence lifetimes and intensities of dyes with different modes of binding to DNA.
AuthorsSailer BL, Nastasi AJ, Valdez JG, Steinkamp JA, Crissman HA
JournalJ Histochem Cytochem
PubMed ID9016307
'Deuterium oxide (D2O) increases both the fluorescence lifetime and the fluorescence intensity of the intercalating dyes propidium iodide (PI) and ethidium bromide (EB) when bound to nucleic acid structures. We have used spectroscopic analysis coupled with conventional and phase-sensitive flow cytometry to compare the alterations in intensity and lifetime of ... More
The repetitive calcium waves in the fertilized ascidian egg are initiated near the vegetal pole by a cortical pacemaker.
AuthorsSpeksnijder JE
JournalDev Biol
PubMed ID1397683
'Ascidian eggs respond to fertilization with a series of repetitive calcium waves that originate mostly from the vegetal/contraction pole region (J. E. Speksnijder, C. Sardet, and L. F. Jaffe, 1990, Dev. Biol. 142, 246-249), where the myoplasm is concentrated during the first phase of ooplasmic segregation. This suggests that the ... More
A rapid method for measuring apoptosis and dual-color immunofluorescence by single laser flow cytometry.
AuthorsSchmid I, Uittenbogaart CH, Keld B, Giorgi JV
JournalJ Immunol Methods
PubMed ID8157993
'A sensitive method for quantification of cells undergoing apoptosis that permits the simultaneous measurement of dual-color cell surface immunofluorescence is presented. Unfixed cells are stained with 7-amino-actinomycin D (7-AAD) for discrimination of live from early apoptotic cells and from cells which have lost membrane integrity (late apoptotic or necrotic, dead ... More
Effects of supravital fluorochromes used to analyze the in vivo homing of murine lymphocytes on cellular function.
AuthorsSamlowski WE, Robertson BA, Draper BK, Prystas E, McGregor JR
JournalJ Immunol Methods
PubMed ID1960398
'A number of supravital fluorochromes are available to study lymphocyte homing in vivo. These include fluorescein isothiocyanate (FITC), which binds to cell surface proteins; Hoechst 33342, which binds to AT rich regions of cellular DNA; and the lipid bilayer incorporated dyes PKH-2 and PKH-26. The relative advantages and disadvantages of ... More
Marginal blebbing during the early stages of TNF-induced apoptosis indicates alteration in actomyosin contractility.
AuthorsDomnina LV, Ivanova OY, Pletjushkina OY, Fetisova EK, Chernyak BV, Skulachev VP, Vasiliev JM
JournalCell Biol Int
PubMed ID15223024
'Dynamics of alterations of cell surface topography during TNF-induced apoptosis of HeLa cells was examined by phase-contrast videomicroscopy and immunomorphological analysis. The final stage of apoptosis accompanied by cell rounding and general blebbing of the cell surface became after 4-6 h of incubation but much earlier, after 1.5-3 h, essentially ... 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
Protection from apoptosis in human neutrophils is determined by the surface of adhesion.
AuthorsGinis I, Faller DV
JournalAm J Physiol
PubMed ID9038836
'Recent work suggests that various neutrophil agonists affect the rate of apoptosis in these cells. On the basis of these observations, we hypothesized that signals triggered in neutrophils via their adhesion receptors might also modify their life span. This hypothesis has been tested using human neutrophils adherent to tissue culture ... More
Imaging of cAMP-dependent protein kinase activity in living neural cells using a novel fluorescent substrate.
AuthorsHigashi H, Sato K, Ohtake A, Omori A, Yoshida S, Kudo Y
JournalFEBS Lett
PubMed ID9305731
'In order to visualize the activity of the cAMP-dependent protein kinase (PKA) in living cells, we have constructed a new fluorescence PKA substrate by conjugating a fluorescence probe to a partial amino acid sequence of PKA regulatory domain II which contains a specific autophosphorylation site. The fluorescent peptide was cell-permeable ... More
PLA2 activity is required for nuclear shrinkage in caspase-independent cell death.
AuthorsShinzawa K, Tsujimoto Y
JournalJ Cell Biol
PubMed ID14676306
'Apoptosis is defined on the basis of morphological changes like nuclear fragmentation and chromatin condensation, which are dependent on caspases. Many forms of caspase-independent cell death have been reported, but the mechanisms are still poorly understood. We found that hypoxic cell death was independent of caspases and was associated with ... More
Fluorogenic label for biomolecular imaging.
AuthorsLavis LD, Chao TY, Raines RT
JournalACS Chem Biol
PubMed ID17163679
'Traditional small-molecule fluorophores are always fluorescent. This attribute can obscure valuable information in biological experiments. Here, we report on a versatile "latent" fluorophore that overcomes this limitation. At the core of the latent fluorophore is a derivative of rhodamine in which one nitrogen is modified as a urea. That modification ... More
Nucleic acid dyes for detection of apoptosis in live cells.
AuthorsFrey T
JournalCytometry
PubMed ID8582249
'Apoptotic thymocytes were found to be much dimmer than normal thymocytes when stained with several nucleic acid dyes. These dyes provide a quick and simple assay for apoptosis which works for live cells and does not require a UV laser. The collection of dyes giving this staining pattern includes reagents ... More
Endocytosis of gastrin in cancer cells expressing gastrin/CCK-B receptor.
AuthorsTarasova NI, Wank SA, Hudson EA, Romanov VI, Czerwinski G, Resau JH, Michejda CJ
JournalCell Tissue Res
PubMed ID8995203
'Endocytosis of gastrin was studied in a number of gastrin-receptor-expressing cell lines by confocal laser scanning microscopy (CLSM) with the aid of a biologically active fluorescent derivative, rhodamine green heptagastrin. Rapid clustering (within 4-7 min) and internalization of fluorescent ligand upon binding at room temperature and 37 degrees C were ... More
A novel nozzle for more efficient sperm orientation to improve sorting efficiency of X and Y chromosome-bearing sperm.
AuthorsRens W, Welch GR, Johnson LA
JournalCytometry
PubMed ID9845443
'Efficient high-resolution detection of DNA for flow cytometric sorting of X and Y chromosome-bearing sperm is dependent on effectively orientating the sperm head to the laser beam in orthogonally configured flow systems. Normally, a beveled needle is required to enlarge the fraction of properly orientated sperm (flat side facing the ... More
Sensitive method for measuring apoptosis and cell surface phenotype in human thymocytes by flow cytometry.
AuthorsSchmid I, Uittenbogaart CH, Giorgi JV
JournalCytometry
PubMed ID7512891
'A rapid, gentle, and sensitive method for quantification of cells undergoing apoptosis is presented. The method allows the simultaneous determination of dual-color cell surface immunofluorescence. Cells are stained for 7 min with the vital dye Hoechst 33342 (HO342) for identification of live and apoptotic cells. 7-amino-actinomycin D (7-AAD) is added ... More
Restrained torsional dynamics of nuclear DNA in living proliferative mammalian cells.
AuthorsTramier M, Kemnitz K, Durieux C, Coppey J, Denjean P, Pansu RB, Coppey-Moisan M
JournalBiophys J
PubMed ID10777758
'Physical parameters, describing the state of chromatinized DNA in living mammalian cells, were revealed by in situ fluorescence dynamic properties of ethidium in its free and intercalated states. The lifetimes and anisotropy decays of this cationic chromophore were measured within the nuclear domain, by using the ultra-sensitive time-correlated single-photon counting ... More
Functional study of multidrug resistance with fluorescent dyes. Limits of the assay for low levels of resistance and application in clinical samples.
AuthorsCanitrot Y, Lahmy S, Buquen JJ, Canitrot D, Lautier D
JournalCancer Lett
PubMed ID8827047
'Fluorescent dyes such as rhodamine 123 (R123) and Hoechst 33342 (Ho342) have been widely used to characterize multidrug-resistance (MDR) phenotype cells in cell populations, on the basis of their reduced accumulation in resistant cells. Taking advantage of the high fluorescence quantum yield of R123 and Ho342 compared with that of ... More
Intracellular pH and the increase in protein synthesis accompanying activation of Xenopus eggs.
AuthorsGrandin N, Charbonneau M
JournalBiol Cell
PubMed ID2620165
'Metabolic activation following egg fertilization corresponds to an increase in protein synthesis and the initiation of DNA synthesis, which lead to cell division and development of the embryo. Since in several biological systems protein synthesis is regulated by intracellular pH (pHi), we have decided to investigate the situation during Xenopus ... More
Hematopoietic stem cell characterization by Hoechst 33342 and rhodamine 123 staining.
AuthorsBertoncello I, Williams B
JournalMethods Mol Biol
PubMed ID14976367
'A dual-dye efflux strategy utilizing the supravital dyes Hoechst 33342 (Ho) and rhodamine 123 (Rh123) is described and illustrated for the detection and analysis of hematopoietic stem cells in murine bone marrow. Mononuclear cells from bone marrow cell suspensions were incubated in a cocktail of Rh123 plus Ho, and both ... More
Adaptation of a simple flow cytometric assay to identify different stages during apoptosis.
AuthorsHamel W, Dazin P, Israel MA
JournalCytometry
PubMed ID8891447
'Apoptosis occurs under many different physiological and pathological conditions, and it reflects a genetically encoded suicide program that can be triggered by different stimuli in susceptible cells. We adapted a flow cytometric assay for the detection of apoptosis based on differential staining of viable cells with two different DNA binding ... More
Verapamil and cyclosporin A sensitize human kidney tumor cells to vincristine in absence of membrane P-glycoprotein and without apparent changes in the cytoplasmic free Ca2+ concentration.
AuthorsNygren P, Larsson R
JournalBiosci Rep
PubMed ID1972632
'Vincristine (Vcr) dose dependently inhibited growth of the kidney adenocarcinoma cell line ACHN during 4 days of culture. Verapamil (Ver) at 10 microM and cyclosporin A (CsA) at 1 microgram/ml had no effect on cell growth but significantly potentiated the action of Vcr, despite the absence of the multidrug resistance ... More
Erythropoiesis in the absence of adult hemoglobin.
AuthorsLiu S, McConnell SC, Ryan TM
JournalMol Cell Biol
PubMed ID23530053
'During erythropoiesis, hemoglobin (Hb) synthesis increases from early progenitors to mature enucleated erythrocytes. Although Hb is one of the most extensively studied proteins, the role of Hb in erythroid lineage commitment, differentiation, and maturation remains unclear. In this study, we generate mouse embryos and embryonic stem (ES) cells with all ... More
YOPRO-1 permits cytofluorometric analysis of programmed cell death (apoptosis) without interfering with cell viability.
AuthorsIdziorek T, Estaquier J, De Bels F, Ameisen JC
JournalJ Immunol Methods
PubMed ID7561136
'In the absence of cell permeabilization, the impermeant nuclear dye YOPRO-1 permits accurate analysis of apoptosis using cytofluorometry or fluorescent microscopy. Several immune cell populations were studied including dexamethasone-treated thymocytes, irradiated peripheral blood mononuclear cells and a growth factor-depleted tumor B cell line. Excellent correlation values were found with acridine ... More
Nucleic acid binding agents exert local toxic effects on neurites via a non-nuclear mechanism.
AuthorsPin S, Chen H, Lein PJ, Wang MM
JournalJ Neurochem
PubMed ID16441515
'The mechanism by which drugs that target nucleic acids cause neurotoxicity is not well described. We characterized the neurotoxicity of Hoechst 33342 (bis-benzimide), a common cell permeable nuclear dye, in primary neuronal cultures. The mechanism of cell death was not apoptotic, as death is rapid, not accompanied by typical nuclear ... More
Cross-talk between calpain and caspase proteolytic systems during neuronal apoptosis.
AuthorsNeumar RW, Xu YA, Gada H, Guttmann RP, Siman R
JournalJ Biol Chem
PubMed ID12576481
'Cross-talk between calpain and caspase proteolytic systems has complicated efforts to determine their distinct roles in apoptotic cell death. This study examined the effect of overexpressing calpastatin, the specific endogenous calpain inhibitor, on the activity of the two proteolytic systems following an apoptotic stimulus. Human SH-SY5Y neuroblastoma cells were stably ... More
Direct observation in the millisecond time range of fluorescent molecule asymmetrical interaction with the electropermeabilized cell membrane.
AuthorsGabriel B, Teissié J
JournalBiophys J
PubMed ID9370457
'Interaction of two stains (propidium iodide and ethidium bromide) with electropermeabilized living Chinese hamster ovary cells is observed using an ultrafast fluorescence image acquisition system. The computing process is linked to an ultra-low-light intensifying camera working with a very short time resolution (3.33 ms per image). Altered parts of the ... More
Fluorescence in situ hybridization of scarce leptin receptor mRNA using the enzyme-labeled fluorescent substrate method and tyramide signal amplification.
AuthorsBreininger JF, Baskin DG
JournalJ Histochem Cytochem
PubMed ID11101627
'To increase the sensitivity of fluorescence in situ hybridization (FISH) for detection of low-abundance mRNAs, we performed FISH on cryostat sections of rat hypothalamus with biotin-labeled riboprobes to leptin receptor (ObRb) and amplified the signal by combining tyramide signal amplification (TSA) and Enzyme-Labeled Fluorescent alkaline phosphatase substrate (ELF) methods. First, ... More
Constitutive expression of the machinery for programmed cell death.
AuthorsWeil M, Jacobson MD, Coles HS, Davies TJ, Gardner RL, Raff KD, Raff MC
JournalJ Cell Biol
PubMed ID8655578
'In the presence of cycloheximide (CHX) to inhibit protein synthesis, a high concentration of staurosporine (STS) induces almost all cells in explant cultures of 8/8 types of newborn mouse organs and 3/3 types of adult mouse organs to die with the characteristic features of apoptosis. Eggs and blastomeres also die ... More
Compartmentalized megakaryocyte death generates functional platelets committed to caspase-independent death.
AuthorsClarke MC, Savill J, Jones DB, Noble BS, Brown SB
JournalJ Cell Biol
PubMed ID12591916
'Caspase-directed apoptosis usually fragments cells, releasing nonfunctional, prothrombogenic, membrane-bound apoptotic bodies marked for rapid engulfment by macrophages. Blood platelets are functional anucleate cells generated by specialized fragmentation of their progenitors, megakaryocytes (MKs), but committed to a constitutive caspase-independent death. Constitutive formation of the proplatelet-bearing MK was recently reported to be ... More
Correlating cell cycle with metabolism in single cells: combination of image and metabolic cytometry.
AuthorsKrylov SN, Zhang Z, Chan NW, Arriaga E, Palcic MM, Dovichi NJ
JournalCytometry
PubMed ID10451502
'BACKGROUND: We coin two terms: First, chemical cytometry describes the use of high-sensitivity chemical analysis techniques to study single cells. Second, metabolic cytometry is a form of chemical cytometry that monitors a cascade of biosynthetic and biodegradation products generated in a single cell. In this paper, we describe the combination ... More
Guidelines for the use and interpretation of assays for monitoring cell death in higher eukaryotes.
AuthorsGalluzzi L, Aaronson SA, Abrams J, Alnemri ES, Andrews DW, Baehrecke EH, Bazan NG, Blagosklonny MV, Blomgren K, Borner C, Bredesen DE, Brenner C, Castedo M, Cidlowski JA, Ciechanover A, Cohen GM, De Laurenzi V, De Maria R, Deshmukh M, Dynlacht BD, El-Deiry WS, Flavell RA, Fulda S, Garrido C, Golstein P, Gougeon ML, Green DR, Gronemeyer H, Hajnóczky G, Hardwick JM, Hengartner MO, Ichijo H, Jäättelä M, Kepp O, Kimchi A, Klionsky DJ, Knight RA, Kornbluth S, Kumar S, Levine B, Lipton SA, Lugli E, Ma
JournalCell Death Differ
PubMed ID19373242
'Cell death is essential for a plethora of physiological processes, and its deregulation characterizes numerous human diseases. Thus, the in-depth investigation of cell death and its mechanisms constitutes a formidable challenge for fundamental and applied biomedical research, and has tremendous implications for the development of novel therapeutic strategies. It is, ... More
Mapping mechanisms and charting the time course of premature cell senescence and apoptosis: lysosomal dysfunction and ganglioside accumulation in endothelial cells.
AuthorsPatschan S, Chen J, Gealekman O, Krupincza K, Wang M, Shu L, Shayman JA, Goligorsky MS,
JournalAm J Physiol Renal Physiol
PubMed ID17928415
'Endothelial cells subjected to glycated collagen I develop premature senescence within 3-5 days, as revealed by increased senescence-associated beta-galactosidase activity, decreased proliferation, and an increase in cell size. Here, we analyzed the time course and possible mechanisms of this process. Lysosomal integrity studies revealed a rapid collapse of pH gradient ... More
Modulation of chondrocyte proliferation by ascorbic acid and BMP-2.
AuthorsVenezian R, Shenker BJ, Datar S, Leboy PS
JournalJ Cell Physiol
PubMed ID9462695
'Chondrocytes show an unusual ability to thrive under serum-free conditions as long as insulin, thyroxine, and cysteine are present. Studies with sternal chondrocytes from chick embryos indicate that thymidine incorporation in chondrocytes cultured under serum-free conditions is 30-50% of that seen with fetal bovine serum (FBS). In contrast, skin fibroblast ... More
Time-resolved fluorescence spectroscopy and imaging of DNA labeled with DAPI and Hoechst 33342 using three-photon excitation.
AuthorsLakowicz JR, Gryczynski I, Malak H, Schrader M, Engelhardt P, Kano H, Hell SW
JournalBiophys J
PubMed ID9017187
'We examined the fluorescence spectral properties of the DNA stains DAPI (4'',6-diamidino-2-phenylindole, hydrochloride) and Hoechst 33342 (bis-benzimide, or 2,5''-bi''1H-benzimidazole2''-(4-ethoxyphenyl)-5-(4-methyl-1-piperazi nyl)) with two-photon (2h nu) and three-photon (3h nu) excitation using femtosecond pulses from a Ti:sapphire laser from 830 to 885 nm. The mode of excitation of DAPI bound to DNA ... 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
Cigarette smoke extract induces oxidative stress and apoptosis in human lung fibroblasts.
AuthorsCarnevali S, Petruzzelli S, Longoni B, Vanacore R, Barale R, Cipollini M, Scatena F, Paggiaro P, Celi A, Giuntini C
JournalAm J Physiol Lung Cell Mol Physiol
PubMed ID12547733
'Cigarette smoke is a mixture of chemicals having direct and/or indirect toxic effects on different lung cells. We investigated the effect of cigarette smoke on human lung fibroblasts (HFL-1) oxidation and apoptosis. Cells were exposed to various concentrations (1, 5, and 10%) of cigarette smoke extract (CSE) for 3 h, ... More
Identification and biochemical characterization of an 80 kilodalton GTP-binding/transglutaminase from rabbit liver nuclei.
AuthorsSingh US, Erickson JW, Cerione RA
JournalBiochemistry
PubMed ID7495818
'The primary aim of these studies was to identify and biochemically characterize GTP-binding proteins in the nucleus. We found that an 80 kDa protein was responsible for the majority of the GTP-binding activity detected in rabbit liver nuclear preparations as assayed by photoaffinity labeling with [alpha-32P]GTP. The GTP-binding activity was ... More
Comparative evaluation of several DNA binding dyes in the detection of apoptosis-associated chromatin degradation by flow cytometry.
AuthorsTelford WG, King LE, Fraker PJ
JournalCytometry
PubMed ID1372208
'Mouse thymocytes readily undergo apoptosis-associated DNA degradation upon exposure to glucocorticoids or ionizing radiation. It has been previously shown that flow cytometric cell cycle analysis of propidium iodide-stained apoptotic thymocytes results in the appearance of a distinct cell cycle region (the A0 region) below the G0/G1 region. Cells in this ... More