Detection of 5-bromo-2-deoxyuridine (BrdUrd) incorporation with monoclonal anti-BrdUrd antibody after deoxyribonuclease treatment.
AuthorsTakagi S, McFadden ML, Humphreys RE, Woda BA, Sairenji T
JournalCytometry
PubMed ID8404370
We studied the effects of deoxyribonucleases on the detection of 5-bromo-2-deoxyuridine (BrdUrd) by anti-BrdUrd monoclonal antibodies (mAbs). After DNase I treatment, BrdUrd was detected in cells fixed on slides with the anti-BrdUrd mAbs, B44 and BMC9318. The level of detection related to the degree of DNA digestion. DNA digestion of ... More
Rac1 inhibits myogenic differentiation by preventing the complete withdrawal of myoblasts from the cell cycle.
AuthorsHeller H, Gredinger E, Bengal E
JournalJ Biol Chem
PubMed ID11489882
'The small GTPase protein Rac1 is involved in a wide range of biological processes, yet its role in cell differentiation is mostly unknown. Here we show that Rac1 activity is high in proliferating myoblasts and decreases during the differentiation process. To analyze the involvement of Rac1 in muscle differentiation, different ... More
Androgen-stimulated DNA synthesis and cytoskeletal changes in fibroblasts by a nontranscriptional receptor action.
AuthorsCastoria G, Lombardi M, Barone MV, Bilancio A, Di Domenico M, Bottero D, Vitale F, Migliaccio A, Auricchio F
JournalJ Cell Biol
PubMed ID12743104
'In NIH3T3 cells, 0.001 nM of the synthetic androgen R1881 induces and stimulates association of androgen receptor (AR) with Src and phosphatidylinositol 3-kinase (Pl3-kinase), respectively, thereby triggering S-phase entry. 10 nM R1881 stimulates Rac activity and membrane ruffling in the absence of the receptor-Src-PI3-kinase complex assembly. The antiandrogen Casodex and ... 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
Visualization of replication initiation and elongation in Drosophila.
AuthorsClaycomb JM, MacAlpine DM, Evans JG, Bell SP, Orr-Weaver TL
JournalJ Cell Biol
PubMed ID12403810
'Chorion gene amplification in the ovaries of Drosophila melanogaster is a powerful system for the study of metazoan DNA replication in vivo. Using a combination of high-resolution confocal and deconvolution microscopy and quantitative realtime PCR, we found that initiation and elongation occur during separate developmental stages, thus permitting analysis of ... More
Proliferation of functional hair cells in vivo in the absence of the retinoblastoma protein.
'In mammals, hair cell loss causes irreversible hearing and balance impairment because hair cells are terminally differentiated and do not regenerate spontaneously. By profiling gene expression in developing mouse vestibular organs, we identified the retinoblastoma protein (pRb) as a candidate regulator of cell cycle exit in hair cells. Differentiated and ... More
Retinal ganglion cell genesis requires lakritz, a Zebrafish atonal Homolog.
AuthorsKay JN, Finger-Baier KC, Roeser T, Staub W, Baier H
JournalNeuron
PubMed ID11430806
'Mutation of the zebrafish lakritz (lak) locus completely eliminates the earliest-born retinal cells, the ganglion cells (RGCs). Instead, excess amacrine, bipolar, and Müller glial cells are generated in the mutant. The extra amacrines are found at ectopic locations in the ganglion cell layer. Cone photoreceptors appear unaffected by the mutation. ... More
Immunoseparation and immunodetection of nucleic acids labeled with halogenated nucleotides.
AuthorsHaider SR, Juan G, Traganos F, Darzynkiewicz Z
JournalExp Cell Res
PubMed ID9260920
'A novel methodology for labeling, isolation, and detection of nucleic acids is described. Nucleic acid isolation is based on in vivo or in vitro incorporation of BrU or BrdU to either RNA or DNA, respectively, followed by immunoprecipitation of the labeled nucleic acid utilizing anti-BrdU MoAb, which crossreacts with BrU, ... 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
Deoxyribonucleic acid replication in single cells and chromosomes by immunologic techniques.
AuthorsGratzner HG, Pollack A, Ingram DJ, Leif RC
JournalJ Histochem Cytochem
PubMed ID815428
'Antibodies to 5-bromodeoxyuridine (BrdU) or iododeoxyuridine may be used to identify cells or regions of chromosomes in which de novo deoxyribonucleic acid synthesis has occurred. The antibodies to BrdU were produced in rabbits by injection of the antigen, a conjugate between bovine serum albumin and bromouridine (BrU), or iodouridine. Specific ... More
Treatment with anti-LFA-1 delays the CD8+ cytotoxic-T-lymphocyte response and viral clearance in mice with primary respiratory syncytial virus infection.
AuthorsRutigliano JA, Johnson TR, Hollinger TN, Fischer JE, Aung S, Graham BS
JournalJ Virol
PubMed ID14990720
'Cytotoxic T lymphocytes (CTLs) play an important role in the immune response against respiratory syncytial virus (RSV) infection. The cell surface molecule lymphocyte function-associated antigen 1 (LFA-1) is an important contributor to CTL activation, CTL-mediated direct cell lysis, and lymphocyte migration. In an attempt to determine the role of LFA-1 ... More
Reduced survival of lens epithelial cells in the alphaA-crystallin-knockout mouse.
AuthorsXi JH, Bai F, Andley UP
JournalJ Cell Sci
PubMed ID12584250
'alphaA-Crystallin (alphaA) is a molecular chaperone expressed preferentially in the lens. alphaA transcripts are first detected during the early stages of lens development and its synthesis continues as the lens grows throughout life. alphaA(-/-) mouse lenses are smaller than controls, and lens epithelial cells derived from these mice have diminished ... More
Intensity calibration and automated cell cycle gating for high-throughput image-based siRNA screens of mammalian cells.
AuthorsPoon SS, Wong JT, Saunders DN, Ma QC, McKinney S, Fee J, Aparicio SA,
JournalCytometry A
PubMed ID18698634
'High-content microscopic screening systems are powerful tools for extracting quantitative multiparameter measures from large number of cells under numerous conditions. These systems perform well in applications that monitor the presence of objects, but lack in their ability to accurately estimate object intensities and summarize these findings due to variations in ... More
A new method for introducing double-strand breaks into cellular DNA.
AuthorsLimoli CL, Ward JF
JournalRadiat Res
PubMed ID7683818
'A novel method is used to introduce double-strand breaks into cellular DNA containing controlled levels of 5-bromo-2''-deoxyuridine (BrdU). Chinese hamster V79 cells substituted with BrdU are treated with Hoechst dye #33258 and then exposed to UVA light. Using neutral elution (pH 7.2) the yield of DNA double-strand breaks is found ... More
Application of 5-bromo-2'deoxyuridine as a label for in situ hybridization in chromosome microdissection and painting, and 3' OH DNA end labeling for apoptosis.
AuthorsMühlmann-Díaz MC, Dullea RG, Bedford JS
JournalBiotechniques
PubMed ID8816240
'We have utilized 5-bromo-2''deoxyuridine (BrdU) substituted DNA as a probe for a number of applications including, principally, for chromosome painting by fluorescence in situ hybridization (FISH) but also for DNA end-labeling to detect apoptotic cell death and for filter hybridization. Br-dUTP was used as a substitute for biotin or digoxigenin-dUTP ... More
Paracrine roles of NAD+ and cyclic ADP-ribose in increasing intracellular calcium and enhancing cell proliferation of 3T3 fibroblasts.
AuthorsFranco L, Zocchi E, Usai C, Guida L, Bruzzone S, Costa A, De Flora A
JournalJ Biol Chem
PubMed ID11274199
'CD38 is a bifunctional ectoenzyme synthesizing from NAD(+) (ADP-ribosyl cyclase) and degrading (hydrolase) cyclic ADP-ribose (cADPR), a powerful universal calcium mobilizer from intracellular stores. Recently, hexameric connexin 43 (Cx43) hemichannels have been shown to release cytosolic NAD(+) from isolated murine fibroblasts (Bruzzone, S., Guida, L., Zocchi, E., Franco, L. and ... More
The use of antibody specific for bromodeoxyuridine for the immunofluorescent determination of DNA replication in single cells and chromosomes.
AuthorsGratzner HG, Leif RC, Ingram DJ, Castro A
JournalExp Cell Res
PubMed ID811484
BAC mapping using fluorescence in situ hybridization.
AuthorsChen XN, Korenberg JR
JournalMethods Mol Biol
PubMed ID15020820
Impairment of immunological memory in the absence of MHC despite survival of memory T cells.
AuthorsKassiotis G, Garcia S, Simpson E, Stockinger B
JournalNat Immunol
PubMed ID11836529
The mechanisms by which immunological memory is maintained after infection or vaccination are still a matter of debate. Long-term survival of memory T cells does not require major histocompatibility complex (MHC) contact. We show here that compared with memory CD4+ T cells that maintain contact with MHC class II, memory ... More
Taking cell-matrix adhesions to the third dimension.
AuthorsCukierman E, Pankov R, Stevens DR, Yamada KM
JournalScience
PubMed ID11721053
Adhesions between fibroblastic cells and extracellular matrix have been studied extensively in vitro, but little is known about their in vivo counterparts. Here, we characterized the composition and function of adhesions in three-dimensional (3D) matrices derived from tissues or cell culture. "3D-matrix adhesions" differ from focal and fibrillar adhesions characterized ... More
A gnotobiotic transgenic mouse model for studying interactions between small intestinal enterocytes and intraepithelial lymphocytes.
AuthorsMysorekar IU, Lorenz RG, Gordon JI,
JournalJ Biol Chem
PubMed ID12138109
The mouse intestinal epithelium undergoes continuous renewal throughout life. Intraepithelial lymphocytes (IELs) represent a significant fraction of this epithelium and play an important role in intestinal mucosal barrier function. We have generated a germ-free transgenic mouse model to examine the effects of a genetically engineered proliferative abnormality in the principal ... More
The sub-cellular localization of Sulfolobus DNA replication.
AuthorsGristwood T, Duggin IG, Wagner M, Albers SV, Bell SD,
JournalNucleic Acids Res
PubMed ID22402489
Analyses of the DNA replication-associated proteins of hyperthermophilic archaea have yielded considerable insight into the structure and biochemical function of these evolutionarily conserved factors. However, little is known about the regulation and progression of DNA replication in the context of archaeal cells. In the current work, we describe the generation ... More
Combinatorial roles of olig2 and neurogenin2 in the coordinated induction of pan-neuronal and subtype-specific properties of motoneurons.
AuthorsMizuguchi R, Sugimori M, Takebayashi H, Kosako H, Nagao M, Yoshida S, Nabeshima Y, Shimamura K, Nakafuku M
JournalNeuron
PubMed ID11567615
Distinct classes of neurons are generated at defined times and positions during development of the nervous system. It remains elusive how specification of neuronal identity coordinates with acquisition of pan-neuronal properties. Here we show that basic helix-loop-helix (bHLH) transcription factors Olig2 and Neurogenin2 (Ngn2) play vital roles in the coordinated ... More
Comparison of the labeling efficiency of BrdU, DiI and FISH labeling techniques in bone marrow stromal cells.
AuthorsLi N, Yang H, Lu L, Duan C, Zhao C, Zhao H,
JournalBrain Res
PubMed ID18468584
Cells are generally labeled during in vivo implantation studies enabling the cells to be traced. The relationship between the labeling efficiency and cellular proliferation after transplantation is critical for the interpretation of data obtained by detection of the signals on tissue sections. Here, we compare cellular labeling methods of rat ... More
Inducible gene targeting in the neonatal vasculature and analysis of retinal angiogenesis in mice.
AuthorsPitulescu ME, Schmidt I, Benedito R, Adams RH,
JournalNat Protoc
PubMed ID20725067
The retina is a powerful experimental system for the analysis of angiogenic blood vessel growth in the postnatal organisms. The three-dimensional architecture of the vessel network and processes as diverse as endothelial cell (EC) proliferation, sprouting, perivascular cell recruitment, vessel remodeling or maturation can be investigated at high resolution. The ... More
Spatial distribution and specification of mammalian replication origins during G1 phase.
AuthorsLi F, Chen J, Solessio E, Gilbert DM
JournalJ Cell Biol
PubMed ID12707307
We have examined the distribution of early replicating origins on stretched DNA fibers when nuclei from CHO cells synchronized at different times during G1 phase initiate DNA replication in Xenopus egg extracts. Origins were differentially labeled in vivo versus in vitro to allow a comparison of their relative positions and ... More
High-resolution physical and transcriptional mapping of the autoimmune polyendocrinopathy-candidiasis-ectodermal dystrophy locus on chromosome 21q22.3 by FISH.
AuthorsAaltonen J, Horelli-Kuitunen N, Fan JB, Björses P, Perheentupa J, Myers R, Palotie A, Peltonen L
JournalGenome Res
PubMed ID9267805
Autoimmune-polyendocrinopathy-candidiasis-ectodermal dystrophy (APECED, PGD type I) is an autosomal recessive disease enriched in the Finnish population. Previously, we have assigned APECED to a 2.6-cM interval on chromosome 21q22.3 by linkage analysis in 14 Finnish families. This subtelomeric region of 21q22.3 seems to have sequence features resulting in its under-representation in ... More
MyoD-positive myoblasts are present in mature fetal organs lacking skeletal muscle.
The epiblast of the chick embryo gives rise to the ectoderm, mesoderm, and endoderm during gastrulation. Previous studies revealed that MyoD-positive cells were present throughout the epiblast, suggesting that skeletal muscle precursors would become incorporated into all three germ layers. The focus of the present study was to examine a ... More
ATP regulates the differentiation of mammalian skeletal muscle by activation of a P2X5 receptor on satellite cells.
AuthorsRyten M, Dunn PM, Neary JT, Burnstock G
JournalJ Cell Biol
PubMed ID12135987
ATP is well known for its role as an intracellular energy source. However, there is increasing awareness of its role as an extracellular messenger molecule (Burnstock, 1997). Although evidence for the presence of receptors for extracellular ATP on skeletal myoblasts was first published in 1983 (Kolb and Wakelam), their physiological ... More
Synergistic effects of MAP2 and MAP1B knockout in neuronal migration, dendritic outgrowth, and microtubule organization.
AuthorsTeng J, Takei Y, Harada A, Nakata T, Chen J, Hirokawa N
JournalJ Cell Biol
PubMed ID11581286
MAP1B and MAP2 are major members of neuronal microtubule-associated proteins (MAPs). To gain insights into the function of MAP2 in vivo, we generated MAP2-deficient (map2(-/-)) mice. They developed without any apparent abnormalities, which indicates that MAP2 is dispensable in mouse survival. Because previous reports suggest a functional redundancy among MAPs, ... More
Simultaneous analysis of surface marker expression and cell cycle progression in human peripheral blood mononuclear cells.
AuthorsRosato MT, Jabbour AJ, Ponce RA, Kavanagh TJ, Takaro TK, Hill JP, Poot M, Rabinovitch PS, Faustman EM
JournalJ Immunol Methods
PubMed ID11516753
One method for examining cell cycle kinetics by flow cytometry uses continuous DNA labeling with bromodeoxyuridine (BrdU), a thymidine analogue. Upon incorporation into DNA, BrdU causes stoichiometric quenching of the DNA fluorochrome Hoechst 33258. After counterstaining with a secondary DNA fluorochrome (e.g., ethidium bromide), the analyst can distinguish cells in ... More
The p42/p44 mitogen-activated protein kinase activation triggers p27Kip1 degradation independently of CDK2/cyclin E in NIH 3T3 cells.
AuthorsDelmas C, Manenti S, Boudjelal A, Peyssonnaux C, Eychène A, Darbon JM
JournalJ Biol Chem
PubMed ID11418594
The p42/p44 mitogen-activated protein (MAP) kinase is stimulated by various mitogenic stimuli, and its sustained activation is necessary for cell cycle G(1) progression and G(1)/S transition. G(1) progression and G(1)/S transition also depend on sequential cyclin-dependent kinase (CDK) activation. Here, we demonstrate that MAP kinase inhibition leads to accumulation of ... More
Human age-related cataract and lens epithelial cell death.
AuthorsHarocopos GJ, Alvares KM, Kolker AE, Beebe DC
JournalInvest Ophthalmol Vis Sci
PubMed ID9856780
PURPOSE: To evaluate the importance of lens epithelial cell death in age-related cataract. To determine whether the large percentage of terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick-end labeling (TUNEL)-positive lens epithelial cells previously reported in human capsulotomy specimens results from apoptosis or necrosis. METHODS: Capsulotomy specimens from patients who had undergone ... More
Epidermal growth factor receptor-mediated activation of Stat3 during multistage skin carcinogenesis.
In the present study, we have investigated the possible role of signal transducers and activators of transcription (STATs), particularly Stat3, in mouse skin tumor promotion and multistage carcinogenesis. Stat1, Stat3, and Stat5 were activated in mouse epidermis after treatment with different classes of tumor promoters, including 12-O-tetradecanoylphorbol-13-acetate (TPA), okadaic acid, ... More
Multiple rows of cells behind an epithelial wound edge extend cryptic lamellipodia to collectively drive cell-sheet movement.
AuthorsFarooqui R, Fenteany G
JournalJ Cell Sci
PubMed ID15585576
The mechanism by which epithelial, endothelial and other strongly cell-cell adhesive cells migrate collectively as continuous sheets is not clear, even though this process is crucial for embryonic development and tissue repair in virtually all multicellular animals. Wound closure in Madin-Darby canine kidney (MDCK) epithelial cell monolayers involves Rac GTPase-dependent ... More
Laser scanning cytometer (LSC) analysis of fraction of labelled mitoses (FLM).
AuthorsGorczyca W, Melamed MR, Darzynkiewicz Z
JournalCell Prolif
PubMed ID9051117
In this report we describe the successful application of a novel microscope-based multiparameter laser scanning cytometer (LSC) to measure duration of different phases of cell cycle in HL-60 human leukaemic cell lines by the fraction of labelled mitoses (FLM) method. Exponentially growing cells were harvested after various time intervals following ... More
Differential drug sensitivity conferred by growth status detected in a mixed population of cycling and noncycling cells.
AuthorsBarcellos-Hoff MH, Marton LJ, Deen DF
JournalCancer Res
PubMed ID2340503
The noncycling cell compartment of tumors is considered to be an important target for chemotherapeutic agents; yet, it has been difficult to accurately quantitate its contribution to tumor response because of a lack of methods that can readily discern the relative sensitivities of cycling and noncycling cells. We have used ... More
Control of intestinal stem cell function and proliferation by mitochondrial pyruvate metabolism.
Authors
JournalNat Cell Biol
PubMed ID28812582
Selective Translation of Cell Fate Regulators Mediates Tolerance to Broad Oncogenic Stress.
Authors
JournalCell Stem Cell
PubMed ID32516567
Regulation of Tumor Initiation by the Mitochondrial Pyruvate Carrier.
Authors
JournalCell Metab
PubMed ID31813825
TGF-β-Induced Quiescence Mediates Chemoresistance of Tumor-Propagating Cells in Squamous Cell Carcinoma.
Authors
JournalCell Stem Cell
PubMed ID29100014
Spatial regulation of VEGF receptor endocytosis in angiogenesis.
Authors
JournalNat Cell Biol
PubMed ID23354168
Multiscale cytometry and regulation of 3D cell cultures on a chip.
Authors
JournalNat Commun
PubMed ID28883466
Directed cardiomyocyte differentiation from human pluripotent stem cells by modulating Wnt/β-catenin signaling under fully defined conditions.
Authors
JournalNat Protoc
PubMed ID23257984
Ribosome biogenesis during cell cycle arrest fuels EMT in development and disease.
Authors
JournalNat Commun
PubMed ID31068593
Phase Separation of a PKA Regulatory Subunit Controls cAMP Compartmentation and Oncogenic Signaling.