Roles of Rho-associated kinase in cytokinesis; mutations in Rho-associated kinase phosphorylation sites impair cytokinetic segregation of glial filaments.
AuthorsYasui Y, Amano M, Nagata K, Inagaki N, Nakamura H, Saya H, Kaibuchi K, Inagaki M
JournalJ Cell Biol
PubMed ID9832553
'Rho-associated kinase (Rho-kinase), which is activated by the small GTPase Rho, regulates formation of stress fibers and focal adhesions, myosin fiber organization, and neurite retraction through the phosphorylation of cytoskeletal proteins, including myosin light chain, the ERM family proteins (ezrin, radixin, and moesin) and adducin. Rho-kinase was found to phosphorylate ... More
Intracellular analysis in vivo of different barosensitive bulbospinal neurons in the rat rostral ventrolateral medulla.
AuthorsGranata AR, Kitai ST
JournalJ Neurosci
PubMed ID1729431
'Neurons located in the rostral ventrolateral medulla (RVLM) with projections to the intermediolateral column (IML) in the spinal cord were electrophysiologically characterized and anatomically identified using an intracellular recording technique in vivo. A group of spontaneously active neurons was antidromically activated by electrical stimulation of the IML in the thoracic ... More
Effect of N-terminal alpha-helix formation on the dimerization and intracellular targeting of alanine:glyoxylate aminotransferase.
AuthorsLumb MJ, Drake AF, Danpure CJ
JournalJ Biol Chem
PubMed ID10400689
'The unparalleled peroxisome-to-mitochondrion mistargeting of alanine:glyoxylate aminotransferase (AGT) in the hereditary disease primary hyperoxaluria type 1 is caused by the combined presence of a common Pro11 --> Leu polymorphism and a disease-specific Gly170 --> Arg mutation. The Pro11 --> Leu replacement generates a functionally weak N-terminal mitochondrial targeting sequence (MTS), ... More
Fluorescence in situ hybridization (FISH): DNA probe production and hybridization criteria.
AuthorsNath J, Johnson KL
JournalBiotech Histochem
PubMed ID9554580
'We describe methods for the production of fluorescence in situ hybridization (FISH) probes and the utilization of these probes for the detection of complementary DNA sequences with accuracy and sensitivity for application in both basic research and clinical diagnosis. Due to the frequent use of FISH in many laboratories, it ... More
Centrophilin: a novel mitotic spindle protein involved in microtubule nucleation.
AuthorsTousson A, Zeng C, Brinkley BR, Valdivia MM
JournalJ Cell Biol
PubMed ID1991791
'A novel protein has been identified which may serve a key function in nucleating spindle microtubule growth in mitosis. This protein, called centrophilin, is sequentially relocated from the centromeres to the centrosomes to the midbody in a manner dependent on the mitotic phase. Centrophilin was initially detected by immunofluorescence with ... More
The human lymph node germinal center cell: characterization and isolation by using two-color flow cytometry.
AuthorsWeinberg DS, Ault KA, Gurley M, Pinkus GS
JournalJ Immunol
PubMed ID2943795
'The germinal center of lymphoid tissues is a critical microenvironmental site of B cell activation and differentiation in response to antigenic stimuli. However, characterization of germinal center cells (GCC) in tissue sections has proved technically difficult. Therefore, we have employed two-color flow cytometric analysis of suspended human tonsillar lymphocytes in ... More
Flow cytometric analysis of erythrocyte populations in Tn syndrome blood using monoclonal antibodies to glycophorin A and the Tn antigen.
AuthorsBigbee WL, Langlois RG, Stanker LH, Vanderlaan M, Jensen RH
JournalCytometry
PubMed ID1690628
'Flow cytometric analysis employing monoclonal antibodies to the Tn antigen and glycophorin A was used to characterize the erythrocyte populations present in blood samples from individuals with Tn syndrome. Four monoclonal antibodies specific for the Tn antigen, Gal-NAc monosaccharide, on human erythrocytes were obtained from a fusion of splenocytes from ... More
Correlation of functional properties of human lymphoid cell subsets and surface marker phenotypes using multiparameter analysis and flow cytometry.
AuthorsLanier LL, Engleman EG, Gatenby P, Babcock GF, Warner NL, Herzenberg LA
JournalImmunol Rev
PubMed ID6226586
B lymphocytes express and lose syndecan at specific stages of differentiation.
AuthorsSanderson RD, Lalor P, Bernfield M
JournalCell Regul
PubMed ID2519615
'Lymphopoietic cells require interactions with bone marrow stroma for normal maturation and show changes in adhesion to matrix during their differentiation. Syndecan, a heparan sulfate-rich integral membrane proteoglycan, functions as a matrix receptor by binding cells to interstitial collagens, fibronectin, and thrombospondin. Therefore, we asked whether syndecan was present on ... 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
Temporary disruption of the plasma membrane is required for c-fos expression in response to mechanical stress.
AuthorsGrembowicz KP, Sprague D, McNeil PL
JournalMol Biol Cell
PubMed ID10198070
'Mechanically stressed cells display increased levels of fos message and protein. Although the intracellular signaling pathways responsible for FOS induction have been extensively characterized, we still do not understand the nature of the primary cell mechanotransduction event responsible for converting an externally acting mechanical stressor into an intracellular signal cascade. ... More
Molecular basis for the dual mitochondrial and cytosolic localization of alanine:glyoxylate aminotransferase in amphibian liver cells.
AuthorsHolbrook JD, Danpure CJ
JournalJ Biol Chem
PubMed ID11694530
'To gain further insights into the molecular basis of the evolution of alanine:glyoxylate aminotransferase (AGT) intracellular targeting in vertebrates, we have studied the molecular basis of its dual mitochondrial and cytosolic distribution in amphibian liver cells. The AGT gene in Xenopus laevis encodes a polypeptide of 415 amino acids, which ... More
Expression of Lyt-1 by a subset of B lymphocytes.
AuthorsManohar V, Brown E, Leiserson WM, Chused TM
JournalJ Immunol
PubMed ID6177768
'Using two-color flow cytometry and multiparameter data analysis, we have shown that the IgM bright, large subset of mouse splenic B lymphocytes express Lyt-1. This is not due to B cell uptake of immune complexes of Lyt-1 and antibody from T cells. The IgM bright cells of autoimmune NZB mice ... More
Simultaneous analysis of cell surface antigens, bromodeoxyuridine incorporation and DNA content.
AuthorsHouck DW, Loken MR
JournalCytometry
PubMed ID2866070
'A method has been developed for correlating the presence of cell surface markers with bromodeoxyuridine (BrdUrd) uptake. We have found that paraformaldehyde fixation will maintain immunofluorescent cell surface staining through acid denaturation that is necessary for anti-BrdUrd reactivity to single-stranded DNA. In addition, the forward angle light scattering was maintained, ... More
Quantitative analysis of Fc gamma receptors on murine spleen cell populations by using dual parameter flow cytometry.
AuthorsTitus JA, Finkelman FD, Stephany DA, Jones JF, Segal DM
JournalJ Immunol
PubMed ID6234350
'The expression of Fc gamma R on subsets of mouse spleen cells was examined by dual parameter flow microfluorometry. B cells were detected by labeling them with antibodies against sIgM, sIgD, sIgG, or I-A; essentially all B cells expressed Fc gamma R. The number of Fc gamma R per cell ... More
A versatile means of intracellular labeling: injection of biocytin and its detection with avidin conjugates.
AuthorsHorikawa K, Armstrong WE
JournalJ Neurosci Methods
PubMed ID3146670
'Biocytin is a biotin-lysine complex of low molecular weight containing about 65% biotin, which retains a high affinity for avidin. Since the latter molecule has been conjugated to several histochemical markers, the use of biocytin as an intracellular marker was investigated. Electrodes were filled with a solution of 4-6% biocytin ... More
A new method to detect acrosome-reacted spermatozoa using biotinylated soybean trypsin inhibitor.
AuthorsArts EG, Kuiken J, Jager S
JournalFertil Steril
PubMed ID7926117
'OBJECTIVE: To develop a method to detect acrosome-reacted spermatozoa on human zonae pellucidae using only commercially available reagents and without need for sperm fixation. DESIGN: Sperm head labeling with biotinylated soybean trypsin inhibitor (SBTI-biotin) was compared with results of a known method using fluorescein isothiocyanate (FITC)-conjugated Pisum sativum agglutinin. The ... More
The density and distribution of serotonergic appositions onto identified neurons in the rat rostral ventromedial medulla.
AuthorsPotrebic SB, Mason P, Fields HL
JournalJ Neurosci
PubMed ID7751910
'Neurons in the rostral ventromedial medulla (RVM) contribute to the modulation of nociceptive transmission and to the analgesic effects of opioids. The RVM contains serotonergic terminal arbors, serotonergic neurons and several types of serotonin (5-HT) receptors. Limited evidence suggests that 5-HT acting within RVM decreases nociceptive responsiveness and contributes to ... More
Double-staining of horizontal and amacrine cells by intracellular injection with lucifer yellow and biocytin in carp retina.
AuthorsTeranishi T, Negishi K
JournalNeuroscience
PubMed ID7514770
'Horizontal and amacrine cells in the isolated carp retina were impaled with micropipette electrode, identified by their characteristic light responses, and injected iontophoretically with markers for morphological study. Both Lucifer Yellow CH and biocytin were injected simultaneously. Lucifer Yellow was seen by its own fluorescence while biocytin was visualized by ... More
Commitment to expression of the metalloendopeptidases, collagenase and stromelysin: relationship of inducing events to changes in cytoskeletal architecture.
AuthorsWerb Z, Hembry RM, Murphy G, Aggeler J
JournalJ Cell Biol
PubMed ID3005336
'Agents that alter the morphology of rabbit synovial fibroblasts induce synthesis of the metalloendopeptidases, collagenase and stromelysin. We studied the relationship of cytoskeletal changes to the commitment to expression of these metalloendopeptidases. Cells treated with cytochalasin B (CB) or 12-O-tetradecanoylphorbol-13-acetate rounded, and only cells that had lost their stress fibers ... More
Human neuroblastoma cells acquire regulated secretory properties and different sensitivity to Ca2+ and alpha-latrotoxin after exposure to differentiating agents.
AuthorsSher E, Denis-Donini S, Zanini A, Bisiani C, Clementi F
JournalJ Cell Biol
PubMed ID2544606
'IMR-32 human neuroblastoma cells are unable to release [3H]dopamine in response to secretagogues. However, they express a normal complement of membrane receptors and ion channels which are efficiently coupled to second messenger production. In the present study we took advantage of the ability of this cell line to differentiate in ... More
Demonstration of B-cell maturation in X-linked immunodeficient mice by simultaneous three-colour immunofluorescence.
AuthorsHardy RR, Hayakawa K, Parks DR, Herzenberg LA
JournalNature
PubMed ID6358897
'CBA/N mice carrying the X-linked immune deficiency gene (xid) have fewer splenic B cells than normal CBA mice and are unresponsive to a certain class of antigens. Studies of B-cell surface-marker expression and immune responsiveness have led to the commonly accepted idea that the B cells in adult xid mice ... More
Micrometer dimension derivatization of biosensor surfaces using confocal dynamic patterning.
AuthorsBrooks SA, Ambrose WP, Kuhr WG
JournalAnal Chem
PubMed ID10405614
'Using laser scanning confocal optics in conjunction with avidin/biotin technology, micrometer-sized patterns of biomolecules were fabricated on glassy-carbon and fused-silica surfaces. Photoactive biotin was immobilized using the 325-nm line of a Helium-Cadmium laser, which was focused through a 25x or 100x quartz microscope objective. A three-dimensional piezoelectric micromanipulator was used ... More
Immunofluorescent flow cytometry in N dimensions. The multiplex labeling approach.
AuthorsBuican TN, Hoffmann GW
JournalCell Biophys
PubMed ID2412696
'The problem of the simultaneous use in flow cytometry of N greater than 2 antibodies in conjunction with two fluorochromes was investigated. Theoretical analysis led to a labeling procedure and reconstruction formula that allow N-dimensional labeling distributions to be obtained from two-dimensional fluorescence distributions. The general problem of M greater ... More
Ultrasensitive detection of biomolecules with fluorescent dye-doped nanoparticles.
AuthorsLian W, Litherland SA, Badrane H, Tan W, Wu D, Baker HV, Gulig PA, Lim DV, Jin S
JournalAnal Biochem
PubMed ID15464962
'Fluorescent-labeled molecules have been used extensively for a wide range of applications in biological detection and diagnosis. A new form of highly luminescent and photostable nanoparticles was generated by doping the fluorescent dye tris(2''2-bipyridyl)dichlororuthenium(II)hexahydrate (Rubpy) inside silica material. Because thousands of fluorescent dye molecules are encapsulated in the silica matrix ... More
B-cell subpopulations identified by two-colour fluorescence analysis.
AuthorsHardy RR, Hayakawa K, Haaijman J, Herzenberg LA
JournalNature
PubMed ID7045679
Fluorescence-activated cell sorting: theory, experimental optimization, and applications in lymphoid cell biology.
AuthorsParks DR, Herzenberg LA
JournalMethods Enzymol
PubMed ID6396481
Approaches to multicolor immunofluorescence measurements.
AuthorsHoffman RA
JournalCytometry Suppl
PubMed ID3076363
An automated device for the preparation of complex reagent mixtures. The immunofluorescence tomograph.
AuthorsBuican TN, Hoffmann GW
JournalCell Biophys
PubMed ID2412697
A microcomputer-controlled device was built that automatically prepares small volumes of mixtures of up to eight reagents. The operation of the system is fast, flexible, and reliable, thus making possible the routine use of experimental protocols that require large numbers of small volume reagent samples, each having a different composition. ... More
Quantitative immunofluorescence, anti-ras p21 antibody specificity, and cellular oncoprotein levels.
AuthorsJones PL, O'Hare CM, Bass RA, Rao JY, Hemstreet GP, Hurst RE
JournalBiochem Biophys Res Commun
PubMed ID2138889
A general approach to investigating specificity and saturation of antibodies by quantitative immunofluorescence is applied to monoclonal antibodies generated against p21 or ras oligopeptides to quantify ras p21 oncoprotein in cultured cells. Ras 10, a panreactive mouse monoclonal antibody, appears to be a superior probe for detection of p21 in ... More
Sensitive and high resolution in situ hybridization to human chromosomes using biotin labelled probes: assignment of the human thymocyte CD1 antigen genes to chromosome 1.
AuthorsAlbertson DG, Fishpool R, Sherrington P, Nacheva E, Milstein C
JournalEMBO J
PubMed ID3053166
A method for in situ hybridization originally developed for mapping genes in the nematode, Caenorhabditis elegans has been adapted for high resolution cytological mapping of genes in the human. The probe DNAs are labelled by incorporation of biotin dUTP and the site of hybridization detected by immunofluorescence. For the accurate ... More
Texas Red, a hydrophilic, red-emitting fluorophore for use with fluorescein in dual parameter flow microfluorometric and fluorescence microscopic studies.
AuthorsTitus JA, Haugland R, Sharrow SO, Segal DM
JournalJ Immunol Methods
PubMed ID6806389
The sulfonylchloride derivative of the red-emitting fluorophore, sulforhodamine 101, has been synthesized in order to provide a reagent for coupling to amino groups on proteins and other compounds, and it is now commercially available under the name 'Texas Red'. Texas Red conjugates of antibodies and other proteins have been prepared ... More
The hypophysiotropic neurotensin-immunoreactive neuronal system of the rat brain.
AuthorsMerchenthaler I, Lennard DE
JournalEndocrinology
PubMed ID1954874
The present study describes the distribution of hypophysiotropic neurotensin-immunoreactive (NTi) perikarya in the rat hypothalamus. After peripheral administration of the retrograde tracer Fluoro-Gold, NT immunoreactivity was demonstrated with fluorescence immunocytochemistry using Texas red-labeled avidin. The results indicate that approximately 70% of all NTi neurons that are connected to the hypophysial ... More
Direct effects of radiation on the avidin-biotin system. Absence of energy transfer.
AuthorsKempner ES, Miller JH
JournalJ Biol Chem
PubMed ID2203786
Frozen solutions of biotinylated glucose-6-phosphate dehydrogenase and fluorescently tagged avidin were exposed to high energy ionizing radiation. Parallel experiments with peroxidase coupled to streptavidin and with biotinylated phycoerythrin were also performed. The loss of function of each compound was analyzed according to target theory. Target analysis revealed that the radiation-sensitive ... More
Expression of muscle genes in heterokaryons depends on gene dosage.
AuthorsPavlath GK, Blau HM
JournalJ Cell Biol
PubMed ID3941151
We report that gene dosage, or the ratio of nuclei from two cell types fused to form a heterokaryon, affects the time course of differentiation-specific gene expression. The rate of appearance of the human muscle antigen, 5.1H11, is significantly faster in heterokaryons with equal or near-equal numbers of mouse muscle ... More
Properties of the kinetochore in vitro. I. Microtubule nucleation and tubulin binding.
AuthorsMitchison TJ, Kirschner MW
JournalJ Cell Biol
PubMed ID4030893
We have isolated chromosomes from Chinese hamster ovary cells arrested in mitosis with vinblastine and examined the interactions of their kinetochores with purified tubulin in vitro. The kinetochores nucleate microtubule (MT) growth with complex kinetics. After an initial lag phase, MTs are continuously nucleated with both plus and minus ends ... More
Rheumatoid factor secretion from human Leu-1+ B cells.
AuthorsHardy RR, Hayakawa K, Shimizu M, Yamasaki K, Kishimoto T
JournalScience
PubMed ID3105057
A human B cell subpopulation identifiable by the expression of the cell surface antigen Leu-1 (CD5) is responsible for most of the immunoglobulin M rheumatoid factor secreted in vitro after the cells are stimulated with Staphylococcus aureus. The ability of B cells bearing the Leu-1 marker (Leu-1+) to secrete rheumatoid ... More
Detection of bcr-abl fusion in chronic myelogeneous leukemia by in situ hybridization.
AuthorsTkachuk DC, Westbrook CA, Andreeff M, Donlon TA, Cleary ML, Suryanarayan K, Homge M, Redner A, Gray J, Pinkel D
JournalScience
PubMed ID2237408
Chronic myelogeneous leukemia (CML) is genetically characterized by fusion of the bcr and abl genes on chromosomes 22 and 9, respectively. In most cases, the fusion involves a reciprocal translocation t(9;22)(q34;q11), which produces the cytogenetically distinctive Philadelphia chromosome (Ph1). Fusion can be detected by Southern (DNA) analysis or by in ... More
Interphase and metaphase resolution of different distances within the human dystrophin gene.
AuthorsLawrence JB, Singer RH, McNeil JA
JournalScience
PubMed ID2203143
Fluorescence in situ hybridization makes possible direct visualization of single sequences not only on chromosomes, but within decondensed interphase nuclei, providing a potentially powerful approach for high-resolution (1 Mb and below) gene mapping and the analysis of nuclear organization. Interphase mapping was able to extend the ability to resolve and ... More
Localization of analyte molecules in MALDI preparations by confocal laser scanning microscopy.
AuthorsHorneffer V, Forsmann A, Strupat K, Hillenkamp F, Kubitscheck U
JournalAnal Chem
PubMed ID11289411
In this study, the incorporation of Texas Red-labeled avidin into crystals of 2,5-dihydroxybenzoic acid (2,5-DHB) and 2,6-DHB (used as matrixes for matrix-assisted laser desorption/ionization (MALDI)) was investigated by fluorescence spectrophotometry and confocal laser scanning microscopy (CLSM). The analyte distribution in crystals, grown slowly under controlled conditions, was compared to the ... More
Poly(A) RNA codistribution with microfilaments: evaluation by in situ hybridization and quantitative digital imaging microscopy.
AuthorsTaneja KL, Lifshitz LM, Fay FS, Singer RH
JournalJ Cell Biol
PubMed ID1360014
The distribution of poly(A) RNA has been visualized in single cells using high-resolution fluorescent in situ hybridization. Digital imaging microscopy was used to quantitate the signal in various cellular compartments. Most of the poly(A) signal remained associated with the cellular filament systems after solubilization of membranes with Triton, dissociation of ... More
Coexpression of T4 and T8 on peripheral blood T cells demonstrated by two-color fluorescence flow cytometry.
AuthorsBlue ML, Daley JF, Levine H, Schlossman SF
JournalJ Immunol
PubMed ID2982943
Using two-color fluorescence flow cytometry, we were able to detect the presence of small numbers of T4+T8+ cells (about 3%) in freshly isolated peripheral T cell populations derived from normal healthy donors. Coexpression of T4 and T8 was predominantly found on large blastlike cells and appeared to be related to ... More
Segregation of micrometer-dimension biosensor elements on a variety of substrate surfaces.
AuthorsBrooks SA, Dontha N, Davis CB, Stuart JK, O'Neill G, Kuhr WG
JournalAnal Chem
PubMed ID10939396
With the rapid development of micro total analysis systems and sensitive biosensing technologies, it is often desirable to immobilize biomolecules to small areas of surfaces other than silicon. To this end, photolithographic techniques were used to derivatize micrometer-sized, spatially segregated biosensing elements on several different substrate surfaces. Both an interference ... More
Fluorescent and biotinylated linear peptides as selective bifunctional ligands for the V1a vasopressin receptor.
AuthorsHowl J, Wang X, Kirk CJ, Wheatley M
JournalEur J Biochem
PubMed ID8477743
We have designed and synthesized a linear peptide analogue of arginine vasopressin. This peptide, [1-phenylacetyl, 2-O-methyl-D-tyrosine, 6-arginine, 8-arginine, 9-lysinamide]vasopressin (PhAcALVP), has a lysinamide residue substituted for the more usual glycinamide at position 9. Derivatization of PhAcALVP at the N epsilon-lysyl amino group with N-hydroxysuccinimide esters of aminomethylcoumarin (Mec) and biotin ... More
Identification of a novel nuclear domain.
AuthorsAscoli CA, Maul GG
JournalJ Cell Biol
PubMed ID1999457
For most known nuclear domains (ND), specific functions have been identified. In this report we used murine mAbs and human autoantibodies to investigate precisely circumscribed structures 0.2-0.3 micron in diameter which appear as "nuclear dots" distributed throughout the nucleoplasm. Nuclear dots are metabolically stable and resistant to nuclease digestion and ... More
Dual fluorochrome analysis of human B lymphocytes: phenotypic examination of resting, anti-immunoglobulin stimulated, and in vivo activated B cells.
B cell-enriched preparations were prepared from human peripheral blood and lymphoid tissues by the depletion of T cells and monocytes. Only B cells by virtue of their staining with anti-B1 conjugated to fluorescein were additionally examined. Dual fluorescence staining and flow cytometric analysis demonstrated that the majority of "resting" human ... More
Rearrangement and expression of endogenous immunoglobulin genes occur in many murine B cells expressing transgenic membrane IgM.
AuthorsStall AM, Kroese FG, Gadus FT, Sieckmann DG, Herzenberg LA, Herzenberg LA
JournalProc Natl Acad Sci U S A
PubMed ID3130629
Transgenic mice carrying immunoglobulin genes coding for mu heavy chain and kappa light chain have been used to study the mechanisms involved in allelic and isotypic exclusion. We report here that individual cells from transgenic mice carrying a functionally rearranged mu heavy chain gene (capable of generating both membrane and ... More
Human lymphocyte subpopulations identified by using three-color immunofluorescence and flow cytometry analysis: correlation of Leu-2, Leu-3, Leu-7, Leu-8, and Leu-11 cell surface antigen expression.
AuthorsLanier LL, Loken MR
JournalJ Immunol
PubMed ID6361119
Three-color immunofluorescence has been used to determine the co-expression of cell surface antigens on human peripheral blood lymphocytes. Monoclonal antibodies or avidin were coupled to either FITC (green), phycoerythrin (orange), or Texas Red (red) fluorochromes. These three fluorochromes could be independently measured by using a dual laser FACS IV system ... More
The mechanism of intercellular aggregation. I. The kinetics of the Fc gamma receptor-mediated aggregation of P388D1 cells with antibody-coated lymphocytes at 4 degrees C.
AuthorsSegal DM, Stephany DA
JournalJ Immunol
PubMed ID6230397
The formation of specific, heterophilic conjugates between cells from the P388D1 mouse macrophage line and antibody-coated mouse spleen cells was followed in cell suspensions at 4 degrees C by dual parameter flow cytometry. Intercellular aggregation in this system is mediated by the binding of the Fc portions of IgG antibodies ... More
Ly-1 B-cell clones similar to human chronic lymphocytic leukemias routinely develop in older normal mice and young autoimmune (New Zealand Black-related) animals.
Studies presented here demonstrate that individually expanded clones of murine Ly-1 B cells, perhaps analogous to the expanded neoplastic Leu-1 B-cell clones in human chronic lymphocytic leukemias, are universally detectable in young New Zealand Black (NZB)-related autoimmune mice and in senescent normal mice (greater than 18 months old). These clones ... More
The C2 domain of the ubiquitin protein ligase Nedd4 mediates Ca2+-dependent plasma membrane localization.
AuthorsPlant PJ, Yeger H, Staub O, Howard P, Rotin D
JournalJ Biol Chem
PubMed ID9405440
Neuronal precursor cell-expressed developmentally down-regulated 4 (Nedd4) is a ubiquitin protein ligase (E3) containing a hect domain, 3 or 4 WW domains, and a putative C2 domain. We have recently demonstrated an association between the WW domains of Nedd4 and the proline-rich PY motifs (XPPXY) of the epithelial Na+ channel, ... More
Subpopulations of mouse Lyt-2+ T cells defined by the expression of an Ly-6-linked antigen, B4B2.
AuthorsFichtner AT, Anderson S, Mage MG, Sharrow SO, Thomas CA, Kung JT
JournalJ Immunol
PubMed ID2435788
The production and characterization of a rat mu,kappa monoclonal anti-mouse T cell subset antibody, B4B2, is reported in this paper. B4B2 typing of lymphoid tissues of commonly used inbred mouse strains revealed two types of reactivity patterns. They can be characterized as C57BL/6-like (B6-like) or C3H/He-like (C3H-like). Among B6-like strains, ... More
Imaging of endosome fusion in BHK fibroblasts based on a novel fluorimetric avidin-biotin binding assay.
AuthorsEmans N, Biwersi J, Verkman AS
JournalBiophys J
PubMed ID8527685
A fluorescence assay of in vivo endosome fusion was developed and applied to define the kinetics of endosome fusion in baby hamster kidney (BHK) fibroblasts. The assay is based on an approximately 10-fold enhancement of the green fluorescence of BODIPY-avidin upon biotin binding. The BODIPY-avidin fluorescence enhancement occurred in < ... More
Noradrenergic input to nociceptive modulatory neurons in the rat rostral ventromedial medulla.
Within the rostral ventromedial medulla (RVM), there are two classes of putative pain modulation neurons: ON cells and OFF cells, which respectively burst or pause prior to withdrawal reflexes elicited by noxious stimulation. Alpha-adrenergic agonists injected into the RVM produce changes in the latency of spinal nocifensive reflexes and, when ... More
Hormonal regulation of the polarized function and distribution of Na/H exchange and Na/HCO3 cotransport in cultured mammary epithelial cells.
AuthorsSjaastad MD, Zettl KS, Parry G, Firestone GL, Machen TE
JournalJ Cell Biol
PubMed ID8393012
The time course for development of polarized function and morphological distribution of pH regulatory mechanisms has been examined in a mouse mammary epithelial cell line (31EG4). Monolayers grown on permeable supports had tight junctions when grown 3-4 days in the presence of the lactogenic hormones dexamethasone (D, a synthetic glucocorticoid) ... More
Quantitative measurements of the specificity and kinetics of conjugate formation between cloned cytotoxic T lymphocytes and splenic target cells by dual parameter flow cytometry.
AuthorsPerez P, Bluestone JA, Stephany DA, Segal DM
JournalJ Immunol
PubMed ID3880573
The formation of conjugates between cloned anti-H-2Kb and Dd cytotoxic T lymphocytes (CTL) and splenic target cells has been studied by dual parameter flow cytometry. By varying effector-target combinations and by blocking with anti-MHC class I monoclonal antibodies, we found that the specificity of conjugate formation, in general, paralleled that ... More
Stimulation of beta1 integrins on fibroblasts induces PDGF independent tyrosine phosphorylation of PDGF beta-receptors.
AuthorsSundberg C, Rubin K
JournalJ Cell Biol
PubMed ID8647902
We report that integrin-mediated signaling induces a rapid and transient tyrosine phosphorylation of platelet-derived growth factor (PDGF) beta-receptors in human diploid foreskin AG 1518 fibroblasts. A transient tyrosine phosphorylation of PDGF beta-receptors was evident one and two hours after cells had been plated on collagen type I and fibronectin, as ... More
The centromere-kinetochore complex: a repeat subunit model.
AuthorsZinkowski RP, Meyne J, Brinkley BR
JournalJ Cell Biol
PubMed ID1828250
The three-dimensional structure of the kinetochore and the DNA/protein composition of the centromere-kinetochore region was investigated using two novel techniques, caffeine-induced detachment of unreplicated kinetochores and stretching of kinetochores by hypotonic and/or shear forces generated in a cytocentrifuge. Kinetochore detachment was confirmed by EM and immunostaining with CREST autoantibodies. Electron ... More