The zinc finger protein A20 inhibits TNF-induced NF-kappaB-dependent gene expression by interfering with an RIP- or TRAF2-mediated transactivation signal and directly binds to a novel NF-kappaB-inhibiting protein ABIN.
The zinc finger protein A20 is a tumor necrosis factor (TNF)â and interleukin 1 (IL-1)-inducible protein that negatively regulates nuclear factor-kappa B (NF-κB)âdependent gene expression. However, the molecular mechanism by which A20 exerts this effect is still unclear. We show that A20 does not inhibit TNF- induced nuclear translocation and ... More
In situ polymerase chain reaction amplification of HIV-1 DNA in brain tissue.
AuthorsStrappe PM, Wang TH, McKenzie CA, Lowrie S, Simmonds P, Bell JE
JournalJ Virol Methods
PubMed ID9562406
A direct in situ polymerase chain reaction (IS-PCR) assay is described for the detection of HIV-1 proviral DNA in formalin fixed paraffin embedded brain tissue. Biotin-16-dUTP is incorporated during the PCR process and microwave pretreatment of tissue sections ensures that no non-specific incorporation into damaged or nicked genomic DNA occurs. ... More
Organization of long-range inhibitory connections with rat visual cortex.
AuthorsMcDonald CT, Burkhalter A
JournalJ Neurosci
PubMed ID7678860
We have studied the laminar organization of local long-range inhibitory connections within rat primary visual cortex (area 17) by combining retrograde tracing of nerve cell bodies with glutamic acid decarboxylase immunocytochemistry. While most inhibitory connections are confined to within 0.4 mm of the injection site, a subset of neurons at ... More
Fibronectin is a component of the sodium dodecyl sulfate-insoluble transglutaminase substrate.
AuthorsTyrrell DJ, Sale WS, Slife CW
JournalJ Biol Chem
PubMed ID2897368
Liver plasma membranes contain a morphologically distinct protein complex which serves as a substrate for the plasma membrane-associated transglutaminase. The complex, which appears as a two-dimensional sheet, is insoluble in sodium dodecyl sulfate and reducing agents and has been named SITS for sodium dodecyl sulfate-insoluble transglutaminase substrate (Tyrrell, D. J., ... More
Autoantigens targeted in systemic lupus erythematosus are clustered in two populations of surface structures on apoptotic keratinocytes.
AuthorsCasciola-Rosen LA, Anhalt G, Rosen A
JournalJ Exp Med
PubMed ID7511686
'Systemic lupus erythematosus is a multisystem autoimmune disease in which the autoantibody response targets a variety of autoantigens of diverse subcellular location. We show here that these autoantigens are clustered in two distinct populations of blebs at the surface of apoptotic cells. The population of smaller blebs contains fragmented endoplasmic ... More
Attenuation of astroglial reactivity by interleukin-10.
AuthorsBalasingam V, Yong VW
JournalJ Neurosci
PubMed ID8622125
'Prominent responses that follow brain trauma include the activation of microglia, recruitment of blood-derived macrophages, and astroglial reactivity. Based on evidence that cytokines produced by macrophages/microglia may cause astrocytes to become reactive, the aim of this study was to determine whether astroglial reactivity could be attenuated by interleukin (IL)-10, a ... More
Transendothelial migration of colon carcinoma cells requires expression of E-selectin by endothelial cells and activation of stress-activated protein kinase-2 (SAPK2/p38) in the tumor cells.
AuthorsLaferriere J, Houle F, Taher MM, Valerie K, Huot J
JournalJ Biol Chem
PubMed ID11448946
'Adhesion and migration of tumor cells on and through the vascular endothelium are critical steps of the metastatic invasion. We investigated the roles of E-selectin and of stress-activated protein kinase-2 (SAPK2/p38) in modulating endothelial adhesion and transendothelial migration of HT-29 colon carcinoma cells. Tumor necrosis factor alpha (TNF alpha) strongly ... More
Activation of calpain I converts excitotoxic neuron death into a caspase-independent cell death.
AuthorsLankiewicz S, Marc Luetjens C, Truc Bui N, Krohn AJ, Poppe M, Cole GM, Saido TC, Prehn JH
JournalJ Biol Chem
PubMed ID10828077
'Glutamate receptor overactivation contributes to neuron death after stroke, trauma, and epileptic seizures. Exposure of cultured rat hippocampal neurons to the selective glutamate receptor agonist N-methyl-d-aspartate (300 microm, 5 min) or to the apoptosis-inducing protein kinase inhibitor staurosporine (300 nm) induced a delayed neuron death. In both cases, neuron death ... More
Fluorescein-labeled tyramide strongly enhances the detection of low bromodeoxyuridine incorporation levels.
AuthorsVan Heusden J, de Jong P, Ramaekers F, Bruwiere H, Borgers M, Smets G
JournalJ Histochem Cytochem
PubMed ID9016320
'Immunocytochemical detection of bromodeoxyuridine (BrdU) labeling can be hampered by low BrdU incorporation levels. We describe here an amplification method for weak BrdU immunosignals. The tyramide signal amplification method based on catalyzed reporter deposition (CARD) uses fluorescein-labeled tyramide as a substrate for horseradish peroxidase. The enzyme catalyzes the formation of ... More
Alpha5beta1 integrin controls cyclin D1 expression by sustaining mitogen-activated protein kinase activity in growth factor-treated cells.
'Cyclin D1 expression is jointly regulated by growth factors and cell adhesion to the extracellular matrix in many cell types. Growth factors are thought to regulate cyclin D1 expression because they stimulate sustained extracellular signal-regulated kinase (ERK) activity. However, we show here that growth factors induce transient ERK activity when ... More
The G protein of vesicular stomatitis virus has free access into and egress from the smooth endoplasmic reticulum of UT-1 cells.
AuthorsBergmann JE, Fusco PJ
JournalJ Cell Biol
PubMed ID2155242
'We have investigated the role of the smooth endoplasmic reticulum (SER) of UT-1 cells in the biogenesis of the glycoprotein (G) of vesicular stomatitis virus (VSV). Using immunofluorescence microscopy, we observed the wild type G protein in the SER of infected cells. When these cells were infected with the mutant ... More
Detection of surface and cytoplasmic CD4 on blood monocytes from normal and HIV-1 infected individuals.
AuthorsFilion LG, Izaguirre CA, Garber GE, Huebsh L, Aye MT
JournalJ Immunol Methods
PubMed ID1703191
'CD4 on blood monocytes is generally regarded as being found on a subset of blood monocytes. However, our results show that all monocytes are CD4+ but the number of molecules per cell is lower than T cells. We have performed immunofluorescent (flow cytometry, microscopy) analysis of monocytes from normal donors ... More
BubR1 is essential for kinetochore localization of other spindle checkpoint proteins and its phosphorylation requires Mad1.
AuthorsChen RH
JournalJ Cell Biol
PubMed ID12163471
'The spindle checkpoint delays anaphase onset until all chromosomes have attached properly to the mitotic spindle. Checkpoint signal is generated at kinetochores that are not bound with spindle microtubules or not under tension. Unattached kinetochores associate with several checkpoint proteins, including BubR1, Bub1, Bub3, Mad1, Mad2, and CENP-E. I herein ... More
HEAT repeats mediate plasma membrane localization of Tor2p in yeast.
AuthorsKunz J, Schneider U, Howald I, Schmidt A, Hall MN
JournalJ Biol Chem
PubMed ID10973982
'The subcellular distribution of Tor1p and Tor2p, two phosphatidylinositol kinase homologs and targets of the immunosuppressive drug rapamycin in Saccharomyces cerevisiae, was analyzed. We found that Tor protein is peripherally associated with membranes. Subcellular fractionation and immunofluorescence studies showed that Tor1p and Tor2p associate with the plasma membrane and a ... More
Fluorescence in situ hybridization using horseradish peroxidase-labeled oligodeoxynucleotides and tyramide signal amplification for sensitive DNA and mRNA detection.
Authorsvan de Corput MP, Dirks RW, van Gijlswijk RP, van de Rijke FM, Raap AK
JournalHistochem Cell Biol
PubMed ID9792422
'We have used horseradish peroxidase-labeled 40 mer oligodeoxynucleotides (HRP-ODNs) specific for the human cytomegalovirus immediate early gene (HCMV-IE) and a novel dinitrophenol-tyramide signal amplification reagent (DNP-TSA plus) to evaluate their utility in fluorescence in situ hybridization (FISH). For DNA FISH, single or cocktails of HRP-ODNs were hybridized to metaphase chromosomes ... More
Chemical subdomains within the kinetochore domain of isolated CHO mitotic chromosomes.
AuthorsWordeman L, Steuer ER, Sheetz MP, Mitchison T
JournalJ Cell Biol
PubMed ID1830054
'We have used indirect immunofluorescence in combination with correlative EM to subdivide the mammalian kinetochore into two domains based on the localization of specific antigens. We demonstrate here that the fibrous corona on the distal face of the kinetochore plate contains tubulin (previously shown by Mitchison, T. J., and M. ... More
Expression and partial characterization of kinesin-related proteins in differentiating and adult skeletal muscle.
AuthorsGinkel LM, Wordeman L
JournalMol Biol Cell
PubMed ID11102514
'Using pan-kinesin antibodies to screen a differentiating C2C12 cell library, we identified the kinesin proteins KIF3A, KIF3B, and conventional kinesin heavy chain to be present in differentiating skeletal muscle. We compared the expression and subcellular localization characteristics of these kinesins in myogenic cells to others previously identified in muscle, neuronal, ... More
Enhancement of immunohistochemical detection of HIV-1 p24 antigen in brain by tyramide signal amplification.
AuthorsStrappe PM, Wang TH, McKenzie CA, Lowrie S, Simmonds P, Bell JE
JournalJ Virol Methods
PubMed ID9274823
'Human immunodeficiency virus type 1 (HIV-1) infection of the brain has been demonstrated in formalin fixed, paraffin embedded post-mortem brain tissue (PM) by chromogenic immunohistochemistry for the HIV p24 antigen. The sensitivity of antigen detection is increased significantly by tyramide signal amplification (TSA) compared to the conventional peroxidase labelled Avidin-Biotin ... More
Interaction with telencephalin and the amyloid precursor protein predicts a ring structure for presenilins.
AuthorsAnnaert WG, Esselens C, Baert V, Boeve C, Snellings G, Cupers P, Craessaerts K, De Strooper B
JournalNeuron
PubMed ID11719200
'The carboxyl terminus of presenilin 1 and 2 (PS1 and PS2) binds to the neuron-specific cell adhesion molecule telencephalin (TLN) in the brain. PS1 deficiency results in the abnormal accumulation of TLN in a yet unidentified intracellular compartment. The first transmembrane domain and carboxyl terminus of PS1 form a binding ... More
Neurite extension occurs in the absence of regulated exocytosis in PC12 subclones.
AuthorsLeoni C, Menegon A, Benfenati F, Toniolo D, Pennuto M, Valtorta F
JournalMol Biol Cell
PubMed ID10473636
'We have investigated the process leading to differentiation of PC12 cells. This process is known to include extension of neurites and changes in the expression of subsets of proteins involved in cytoskeletal rearrangements or in neurosecretion. To this aim, we have studied a PC12 clone (trk-PC12) stably transfected with the ... More
MAUB is a new mucin antigen associated with bladder cancer.
AuthorsBergeron A, Champetier S, LaRue H, Fradet Y
JournalJ Biol Chem
PubMed ID8636121
'The M344 tumor-associated antigen, expressed in 70% of superficial bladder tumors, is a sialylated carbohydrate present on a high molecular mass thiol-reducible secreted mucin, which we named MAUB for mucin antigen of the urinary bladder. Herein we studied the relationship between MAUB and other known mucins in the MGH-U3 bladder ... More
Control of mammary epithelial differentiation: basement membrane induces tissue-specific gene expression in the absence of cell-cell interaction and morphological polarity.
AuthorsStreuli CH, Bailey N, Bissell MJ
JournalJ Cell Biol
PubMed ID1955479
'Functional differentiation in mammary epithelia requires specific hormones and local environmental signals. The latter are provided both by extracellular matrix and by communication with adjacent cells, their action being intricately connected in what appears to be a cascade of events leading to milk production. To distinguish between the influence of ... More
Posttranslational modifications of alpha tubulin: detyrosination and acetylation differentiate populations of interphase microtubules in cultured cells.
AuthorsBulinski JC, Richards JE, Piperno G
JournalJ Cell Biol
PubMed ID3283150
'Subsets of microtubules enriched in posttranslationally detyrosinated (Gundersen, G. G., M. H. Kalnoski, and J. C. Bulinski. 1984. Cell. 38:779) or acetylated (Piperno, G., M. Le Dizet, and X. Chang. 1987. J. Cell Biol. 104:298), alpha tubulin have previously been described in interphase cultured cells. In this study an immunofluorescence ... More
Airway epithelial cell migration dynamics. MMP-9 role in cell-extracellular matrix remodeling.
AuthorsLegrand C, Gilles C, Zahm JM, Polette M, Buisson AC, Kaplan H, Birembaut P, Tournier JM
JournalJ Cell Biol
PubMed ID10427102
'Cell spreading and migration associated with the expression of the 92-kD gelatinase (matrix metalloproteinase 9 or MMP-9) are important mechanisms involved in the repair of the respiratory epithelium. We investigated the location of MMP-9 and its potential role in migrating human bronchial epithelial cells (HBEC). In vivo and in vitro, ... More
Biotin staining in the giant fiber systems of the lobster.
AuthorsMa PM
JournalJ Comp Neurol
PubMed ID7515403
'The avidin-biotin-complex method is a popular immunocytochemical technique. This method labels consistently a group of neurons in the lobster ventral nerve cord in the absence of primary antibodies. The specific staining is due to a relatively high level of endogenous biotin (or biocytin) in these neurons. These biotin-positive neurons are ... More
A Drosophila Tpr protein homolog is localized both in the extrachromosomal channel network and to nuclear pore complexes.
AuthorsZimowska G, Aris JP, Paddy MR
JournalJ Cell Sci
PubMed ID9152019
'Here we report structural, molecular, and biochemical characterizations of Bx34, a Drosophila melanogaster nuclear coiled-coil protein which is localized to extrachromosomal and extranucleolar spaces in the nuclear interior and which is homologous to the mammalian nuclear pore complex protein Tpr. In the nuclear interior, Bx34 is excluded from chromosomes and ... More
Lipid-modified, cysteinyl-containing peptides of diverse structures are efficiently S-acylated at the plasma membrane of mammalian cells.
AuthorsSchroeder H, Leventis R, Shahinian S, Walton PA, Silvius JR
JournalJ Cell Biol
PubMed ID8707845
'A variety of cysteine-containing, lipid-modified peptides are found to be S-acylated by cultured mammalian cells. The acylation reaction is highly specific for cysteinyl over serinyl residues and for lipid-modified peptides over hydrophilic peptides. The S-acylation process appears by various criteria to be enzymatic and resembles the S-acylation of plasma membrane-associated ... More
Differential and specific labeling of epithelial and vascular endothelial cells of the rat lung by Lycopersicon esculentum and Griffonia simplicifolia I lectins.
AuthorsBankston PW, Porter GA, Milici AJ, Palade GE
JournalEur J Cell Biol
PubMed ID1879433
'In the rat lung, we found that the Lycopersicon esculentum (LEA) lectin specifically binds to the epithelium of bronchioles and alveoli whereas Griffonia simplicifolia I (GS-I) binds to the endothelium of alveolar capillaries. The differential binding affinity of these lectins was examined on semithin (approximately 0.5 microns) and thin (less ... More
Quantitative studies of changes in cortical granule number and distribution in the mouse oocyte during meiotic maturation.
AuthorsDucibella T, Anderson E, Albertini DF, Aalberg J, Rangarajan S
JournalDev Biol
PubMed ID3141231
'Cortical granules (CGs) undergo a substantial change in distribution in the mouse oocyte cortex during meiotic maturation. In order to determine the mechanism of their change in distribution near the time of ovulation, CG density, total number per oocyte, and domain areas were quantitated. CGs were visualized microscopically by Lens ... More
Notch1 expression in early lymphopoiesis influences B versus T lineage determination.
'Notch receptors regulate fate decisions in many cells. One outcome of Notch signaling is differentiation of bipotential precursors into one cell type versus another. To investigate consequences of Notch1 expression in hematolymphoid progenitors, mice were reconstituted with bone marrow (BM) transduced with retroviruses encoding a constitutively active form of Notch1. ... More
Dynamics of the genome during early Xenopus laevis development: karyomeres as independent units of replication.
AuthorsLemaitre JM, Géraud G, Méchali M
JournalJ Cell Biol
PubMed ID9732278
'During Xenopus laevis early development, the genome is replicated in less than 15 min every 30 min. We show that during this period, DNA replication proceeds in an atypical manner. Chromosomes become surrounded by a nuclear membrane lamina forming micronuclei or karyomeres. This genomic organization permits that prereplication centers gather ... More
Purification and characterization of membrane proteins.
AuthorsKraehenbuhl JP, Bonnard C
JournalMethods Enzymol
PubMed ID2167430
Role of T cells and germinal center formation in the generation of immune responses to the thymus-independent carbohydrate dextran B512.
AuthorsSverremark E, Fernandez C
JournalJ Immunol
PubMed ID9794393
'Immunization with the thymus-independent (TI) Ag native dextran (DX) B512 induces germinal center (GC) formation in the spleen. However, despite this GC formation, the anti-DX response is poor, and no affinity maturation can be observed. Using cholera toxin (CT) as an adjuvant, splenic as well as humoral responses to DX ... More
Modification of annexin II expression in PC12 cell lines does not affect Ca(2+)-dependent exocytosis.
AuthorsGraham ME, Gerke V, Burgoyne RD
JournalMol Biol Cell
PubMed ID9188096
'The Ca2+/phospholipid/cytoskeletal-binding protein annexin II has been proposed to play an important role in Ca(2+)-dependent exocytosis; however, the evidence for this role is inconclusive. More direct evidence obtained by manipulating annexin II levels in cells is still required. We have attempted to do this by generating stably transfected PC12 cell ... More
In vivo self-renewal of c-Kit+ Sca-1+ Lin(low/-) hemopoietic stem cells.
AuthorsOsawa M, Nakamura K, Nishi N, Takahasi N, Tokuomoto Y, Inoue H, Nakauchi H
JournalJ Immunol
PubMed ID8617942
'The long term marrow-repopulating ability of mouse bone marrow c-Kit+ Sca-1+ Lin(low/-) cells was studied. Injection of as few as 100 c-Kit+ Sca-1+ Lin(low/-) cells could rescue a lethally irradiated recipient mouse and reconstitute both myeloid and lymphoid cells in their peripheral blood for at least 10 mo. When limiting ... More
Strategic location of calcium channels at transmitter release sites of frog neuromuscular synapses.
AuthorsRobitaille R, Adler EM, Charlton MP
JournalNeuron
PubMed ID1980068
'The localization of Ca2+ channels relative to the position of transmitter release sites was investigated at the frog neuromuscular junction (NMJ). Ca2+ channels were labeled with fluorescently tagged omega-conotoxin GVIA, an irreversible Ca2+ channel ligand, and observed with a confocal laser scanning microscope. The Ca2+ channel labeling almost perfectly matched ... More
Sensitive double-labeling technique of retrograde biotinized tracer (biotin-WGA) and immunocytochemistry: light and electron microscopic analysis.
AuthorsShiosaka S, Shimada S, Tohyama M
JournalJ Neurosci Methods
PubMed ID2422502
'We developed a simple method to identify fiber projections that contain bioactive substances using biotinized wheat-germ agglutinin. Tracer that accumulated retrogradely in the soma was made visible under the light microscope by linking it with streptavidin-Texas red, which has a red fluorescence, while antigen was demonstrated in the same section ... More
Optical imaging techniques (histochemical, immunohistochemical, and in situ hybridization staining methods) to visualize mitochondria.
AuthorsTanji K, Bonilla E
JournalMethods Cell Biol
PubMed ID11381601
Hapten-mediated immunocytochemistry: the use of fluorescent and nonfluorescent haptens for the study of cytoskeletal dynamics in living cells.
AuthorsGorbsky GJ, Borisy GG
JournalMethods Cell Biol
PubMed ID2643759
Nigrostriatal dopamine neurons receive substance P-ergic inputs in the substantia nigra: application of the immunoelectron microscopic mirror technique to fluorescent double-staining for transmitter-specific projections.
AuthorsKawai Y, Takagi H, Kumoi Y, Shiosaka S, Tohyama M
JournalBrain Res
PubMed ID2434194
A direct synaptic contact between nigrostriatal dopamine neurons and substance P axons in the substantia nigra was demonstrated using the immunoelectron microscopic mirror technique combined with the fluorescent double-staining method for transmitter-specific projections. Substance P-immunoreactive terminals were found to make synaptic contact with nigral cells exhibiting tyrosine hydroxylase immunoreactivity and ... More
Anti-biotin antibodies offer superior organelle-specific labeling of mitochondria over avidin or streptavidin.
AuthorsHollinshead M, Sanderson J, Vaux DJ
JournalJ Histochem Cytochem
PubMed ID9267466
The mitochondrial matrix contains endogenously biotinylated proteins. These proteins can cause unexpected background signal when biotin-avidin- or biotin-streptavidin-based detection systems are used in immunocytochemistry. Here we show that this reactivity can be deliberately exploited, using a simple anti-biotin reagent, to obtain strong and highly specific labeling of mitochondria by both ... More
Simple modification of a commercial flow cytometer to triple laser excitation. Simultaneous five-color fluorescence detection.
AuthorsWoronicz JD, Rice GC
JournalJ Immunol Methods
PubMed ID2661695
We describe a simple and inexpensive modification to the Coulter 753 flow cytometer which enables simultaneous triple laser excitation for advanced multi-color analysis and sorting applications. The salient feature of the modification was to split the rear laser operating in the all lines mode for use in pumping the rhodamine ... More
Thy-1 is a component common to multiple populations of synaptic vesicles.
AuthorsJeng CJ, McCarroll SA, Martin TF, Floor E, Adams J, Krantz D, Butz S, Edwards R, Schweitzer ES
JournalJ Cell Biol
PubMed ID9456327
Thy-1, a glycosylphosphatidylinositol-linked integral membrane protein of the immunoglobulin superfamily, is a component of both large dense-core and small clear vesicles in PC12 cells. A majority of this protein, formerly recognized only on the plasma membrane of neurons, is localized to regulated secretory vesicles. Thy-1 is also present in synaptic ... More
Immunohistochemical methods to detect nitrotyrosine.
AuthorsViera L, Ye YZ, Estévez AG, Beckman JS
JournalMethods Enzymol
PubMed ID9919586
The immunohistochemical detection of nitrotyrosine is a robust method for detecting peroxynitrite and other reactive nitrogen species. Success depends on optimizing conditions for the particular tissue and experimental design under investigation and the use of positive and negative controls to verify specificity. The two controls of dithionite reduction and blocking ... More
Unexpected presence of neurofilaments in axon-bearing horizontal cells of the mammalian retina.
AuthorsPeichl L, González-Soriano J
JournalJ Neurosci
PubMed ID8366362
In several mammals only one of the two types of retinal horizontal cell, the axonless A-type, appears to express neurofilaments. Neurofilament immunostaining of rodent retinas reveals a horizontal cell plexus that has previously been interpreted as belonging to A-type cells. Our intracellular Lucifer yellow injections strongly suggest that there are ... 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
Cysteine string protein functions directly in regulated exocytosis.
AuthorsChamberlain LH, Burgoyne RD
JournalMol Biol Cell
PubMed ID9693380
Cysteine string protein (Csp) is essential for neurotransmitter release in Drosophila. It has been suggested that Csp functions by regulating the activity of presynaptic Ca2+ channels, thus controlling exocytosis. We have examined the effect of overexpressing Csp1 in PC12 cells, a neuroendocrine cell line. PC12 cell clones overexpressing Csp1 did ... More
A role for c-myc in the regulation of thymocyte differentiation and possibly positive selection.
The purpose of thymocyte differentiation is to establish the T cell repertoire and eliminate nonfunctional and autoreactive T cells. In an analysis of potential regulators of this process, we have found that c-myc expression is down-regulated dramatically during early thymocyte differentiation and subsequently up-regulated along with TCR/CD3 in CD4+8+ cells. ... More
DNA replication is required for the checkpoint response to damaged DNA in Xenopus egg extracts.
AuthorsStokes MP, Van Hatten R, Lindsay HD, Michael WM
JournalJ Cell Biol
PubMed ID12213834
Alkylating agents, such as methyl methanesulfonate (MMS), damage DNA and activate the DNA damage checkpoint. Although many of the checkpoint proteins that transduce damage signals have been identified and characterized, the mechanism that senses the damage and activates the checkpoint is not yet understood. To address this issue for alkylation ... More
Apoptosis and delayed degeneration after spinal cord injury in rats and monkeys.
AuthorsCrowe MJ, Bresnahan JC, Shuman SL, Masters JN, Beattie MS
JournalNat Med
PubMed ID8986744
Apoptosis is a morphologically defined form of programmed cell death seen in a variety of circumstances, including immune cell selection, carcinogenesis and development. Apoptosis has very recently been seen after ischemic or traumatic injury to the central nervous system (CNS), suggesting that active cell death as well as passive necrosis ... More
Association of myosin I alpha with endosomes and lysosomes in mammalian cells.
AuthorsRaposo G, Cordonnier MN, Tenza D, Menichi B, Dürrbach A, Louvard D, Coudrier E
JournalMol Biol Cell
PubMed ID10233157
Myosin Is, which constitute a ubiquitous monomeric subclass of myosins with actin-based motor properties, are associated with plasma membrane and intracellular vesicles. Myosin Is have been proposed as key players for membrane trafficking in endocytosis or exocytosis. In the present paper we provide biochemical and immunoelectron microscopic evidence indicating that ... More
Mapping replicational sites in the eucaryotic cell nucleus.
AuthorsNakayasu H, Berezney R
JournalJ Cell Biol
PubMed ID2910875
We have used fluorescent microscopy to map DNA replication sites in the interphase cell nucleus after incorporation of biotinylated dUTP into permeabilized PtK-1 kangaroo kidney or 3T3 mouse fibroblast cells. Discrete replication granules were found distributed throughout the nuclear interior and along the periphery. Three distinct patterns of replication sites ... More
Dual channel confocal laser scanning microscopy of lucifer yellow-microinjected human brain cells combined with Texas red immunofluorescence.
AuthorsBelichenko PV, Dahlström A
JournalJ Neurosci Methods
PubMed ID7967715
A method for visualization of individual human brain cells and their dendritic extensions in combination with immunofluorescence is described. Microinjection of Lucifer Yellow was used to reveal the dendritic morphology of cortical brain cells. Indirect immunofluorescence with Texas Red as label was used to investigate the distribution of 3 different ... More
Simultaneous cytometric analysis for the expression of cytoplasmic and surface antigens in activated T cells.
AuthorsHayden GE, Walker KZ, Miller JF, Wotherspoon JS, Raison RL
JournalCytometry
PubMed ID2842118
A method of two-colour immunofluorescence staining has been developed to allow the simultaneous analysis of both surface and cytoplasmic antigens. This involves the use of direct fluorochrome antibody conjugates for cell-surface antigen staining, followed by cell permeabilization and the staining of cytoplasmic antigens with biotinylated antibodies and streptavidin-fluorochrome conjugates. Fluorochrome-antibody ... More
NF-kappaB inhibits apoptosis in murine mammary epithelia.
AuthorsClarkson RW, Heeley JL, Chapman R, Aillet F, Hay RT, Wyllie A, Watson CJ
JournalJ Biol Chem
PubMed ID10777569
The transcription factor NF-kappaB is a key modulator of apoptosis in a variety of cell types, but to date this specific function of NF-kappaB has not been demonstrated in epithelia. Here, we describe the activation of NF-kappaB during post-lactational involution of the mouse mammary gland, a period of extensive apoptosis ... More
The direct binding of an HIV fragment with granulocytes from healthy subjects and infected patients.
AuthorsGabrilovich DI, Kozich AT, Moshnikov SA, Pokrovsky VV
JournalScand J Immunol
PubMed ID1535992
The binding of the synthetic fragment from the CD4 binding site of HIV (gp 120) with polymorphonuclear neutrophils (PMN) in 13 healthy donors and 31 HIV-infected patients was studied using a biotin-streptavidin-texas-red complex. The largest percentage of PMN-bound peptide was reached at a final peptide concentration of 10 micrograms/ml. The ... More
DNA fiber-FISH staining mechanism.
Authorsvan de Rijke FM, Florijn RJ, Tanke HJ, Raap AK
JournalJ Histochem Cytochem
PubMed ID10820147
Fluorescence in situ hybridization to DNA fibers (Fiber-FISH) is a high-resolution, wide-ranging physical DNA mapping method that finds increasing application in the study of pathological gene rearrangements. Here we present experiments designed to understand the nature of the discontinuous FISH signal patterns seen after Fiber-FISH. Use of a novel cisplatin-based ... More
Identification of two major sites in the type I interleukin-1 receptor cytoplasmic region responsible for coupling to pro-inflammatory signaling pathways.
AuthorsSlack JL, Schooley K, Bonnert TP, Mitcham JL, Qwarnstrom EE, Sims JE, Dower SK
JournalJ Biol Chem
PubMed ID10671496
Type I interleukin-1 receptor is the prototype for a family of proteins, which play a central role in early responses to injury and infection. The similarity of function across the family is reflected in similarity in signaling: all members tested couple to activation of NFkappaB and stress kinases. The coupling ... More
Caspase-2 is localized at the Golgi complex and cleaves golgin-160 during apoptosis.
AuthorsMancini M, Machamer CE, Roy S, Nicholson DW, Thornberry NA, Casciola-Rosen LA, Rosen A
JournalJ Cell Biol
PubMed ID10791974
Caspases are an extended family of cysteine proteases that play critical roles in apoptosis. Animals deficient in caspases-2 or -3, which share very similar tetrapeptide cleavage specificities, exhibit very different phenotypes, suggesting that the unique features of individual caspases may account for distinct regulation and specialized functions. Recent studies demonstrate ... More
Bacterial endosymbiont-derived lipopolysaccharides and a protein on symbiosome membranes in newly infected amoebae and their roles in lysosome-symbiosome fusion.
AuthorsKim KJ, Na YE, Jeon KW
JournalInfect Immun
PubMed ID8262651
Experimental results are presented to support the view that symbiont-derived lipopolysaccharides are involved in the prevention of lysosome-symbiosome fusion in xD amoebae harboring bacterial endosymbionts. Monoclonal antibodies against lipopolysaccharides and a 96-kDa protein present on symbiosome membranes of amoebae were used to monitor the appearance of the membrane-specific components in ... More
Endocytosis and intracellular transport of transferrin across the lactating rabbit mammary epithelial cell.
AuthorsSeddiki T, Delpal S, Ollivier-Bousquet M
JournalJ Histochem Cytochem
PubMed ID1527373
To study the transcytosis and segregation of ligand in the mammary epithelial cell, endocytosis and intracellular transit of human blood transferrin were followed in lactating rabbit mammary epithelial cells. Human transferrin labeled with biotin added to an incubation medium was bound to the basal membrane of mammary epithelial cells and ... More
Patterns of DNA replication in Drosophila polytene nuclei replicating in Xenopus egg and oocyte extracts.
AuthorsSleeman AM, Leno GH, Mills AD, Fairman MP, Laskey RA
JournalJ Cell Sci
PubMed ID1522141
We have used Xenopus laevis cell-free extracts to study patterns of DNA replication in polytene nuclei isolated from salivary glands of Drosophila melanogaster 3rd instar larvae. Replication was visualized by supplementation with biotin-dUTP so that nascent DNA became labelled, thus allowing detection with fluorescein or Texas-Red-conjugated streptavidin. Biotin incorporation was ... More
Dynamic changes in the mobility of LAT in aggregated lipid rafts upon T cell activation.
AuthorsTanimura N, Nagafuku M, Minaki Y, Umeda Y, Hayashi F, Sakakura J, Kato A, Liddicoat DR, Ogata M, Hamaoka T, Kosugi A
JournalJ Cell Biol
PubMed ID12515827
Lipid rafts are known to aggregate in response to various stimuli. By way of raft aggregation after stimulation, signaling molecules in rafts accumulate and interact so that the signal received at a given membrane receptor is amplified efficiently from the site of aggregation. To elucidate the process of lipid raft ... More
Long-term culture of purified postnatal oligodendrocyte precursor cells. Evidence for an intrinsic maturation program that plays out over months.
AuthorsTang DG, Tokumoto YM, Raff MC
JournalJ Cell Biol
PubMed ID10704447
Oligodendrocytes myelinate axons in the vertebrate central nervous system (CNS). They develop from precursor cells (OPCs), some of which persist in the adult CNS. Adult OPCs differ in many of their properties from OPCs in the developing CNS. In this study we have purified OPCs from postnatal rat optic nerve ... More
p150TSP, a conserved nuclear phosphoprotein that contains multiple tetratricopeptide repeats and binds specifically to SH2 domains.
AuthorsMalek SN, Yang CH, Earnshaw WC, Kozak CA, Desiderio S
JournalJ Biol Chem
PubMed ID8636124
Src homology 2 (SH2) domains are structural modules that function in the assembly of multicomponent signaling complexes by binding to specific phosphopeptides. The tetratricopeptide repeat (TPR) is a distinct structural motif that has been suggested to mediate protein-protein interactions. Among SH2-binding phosphoproteins purified from the mouse B cell lymphoma A20, ... More
Multivesicular endosomes containing internalized EGF-EGF receptor complexes mature and then fuse directly with lysosomes.
AuthorsFutter CE, Pearse A, Hewlett LJ, Hopkins CR
JournalJ Cell Biol
PubMed ID8601581
We have followed the transfer of EGF-EGF receptor (EGFR) complexes from endosomal vacuoles that contain transferrin receptors (TfR) to lysosome vacuoles identified by their content of HRP loaded as a 15-min pulse 4 h previously. We show that the HRP-loaded lysosomes are lysosomal-associated membrane protein-1 (LAMP-1) positive, mannose-6-phosphate receptor (M6PR) ... More
The fibronectin-binding integrins alpha5beta1 and alphavbeta3 differentially modulate RhoA-GTP loading, organization of cell matrix adhesions, and fibronectin fibrillogenesis.
AuthorsDanen EH, Sonneveld P, Brakebusch C, Fassler R, Sonnenberg A
JournalJ Cell Biol
PubMed ID12486108
We have studied the formation of different types of cell matrix adhesions in cells that bind to fibronectin via either alpha5beta1 or alphavbeta3. In both cases, cell adhesion to fibronectin leads to a rapid decrease in RhoA activity. However, alpha5beta1 but not alphavbeta3 supports high levels of RhoA activity at ... More
A practical approach to multicolor flow cytometry for immunophenotyping.
AuthorsBaumgarth N, Roederer M
JournalJ Immunol Methods
PubMed ID10986408
Through a series of novel developments in flow cytometry hardware, software, and dye-chemistry it is now possible to simultaneously measure up to 11 distinct fluorescences and two scattered light parameters on each cell. Such advanced multicolor systems have a number of advantages over current two- and three-color flow cytometric measurements. ... More
Induction of cell shape changes through activation of the interleukin-3 common beta chain receptor by the RON receptor-type tyrosine kinase.
AuthorsMera A, Suga M, Ando M, Suda T, Yamaguchi N
JournalJ Biol Chem
PubMed ID10336478
The RON receptor-type tyrosine kinase, a member of the hepatocyte growth factor receptor family, is a receptor for macrophage-stimulating protein (MSP). Recently, we observed that MSP induces morphological changes in interleukin (IL)-3-dependent Ba/F3 cells ectopically expressing RON. We show here that stimulation of those cells with either MSP or IL-3 ... More
Electric field-induced redistribution and postfield relaxation of epidermal growth factor receptors on A431 cells.
AuthorsGiugni TD, Braslau DL, Haigler HT
JournalJ Cell Biol
PubMed ID3494733
The lateral mobility of the epidermal growth factor (EGF) receptor in the plane of the plasma membrane of cultured A431 cells was investigated using direct and indirect fluorescent probes to measure the generation and relaxation of electric field-induced receptor asymmetry. A steady electric field of 15 V/cm for 30 min ... More
Sorting of yeast membrane proteins into an endosome-to-Golgi pathway involves direct interaction of their cytosolic domains with Vps35p.
AuthorsNothwehr SF, Ha SA, Bruinsma P
JournalJ Cell Biol
PubMed ID11038177
Resident late-Golgi membrane proteins in Saccharomyces cerevisiae are selectively retrieved from a prevacuolar-endosomal compartment, a process dependent on aromatic amino acid-based sorting determinants on their cytosolic domains. The formation of retrograde vesicles from the prevacuolar compartment and the selective recruitment of vesicular cargo are thought to be mediated by a ... More
The nuclear membrane determines the timing of DNA replication in Xenopus egg extracts.
AuthorsLeno GH, Laskey RA
JournalJ Cell Biol
PubMed ID1993731
We have exploited a property of chicken erythrocyte nuclei to analyze the regulation of DNA replication in a cell-free system from Xenopus eggs. Many individual demembranated nuclei added to the extract often became enclosed within a common nuclear membrane. Nuclei within such a "multinuclear aggregate" lacked individual membranes but shared ... More
Calcitonin gene-related peptide: possible role in formation and maintenance of neuromuscular junctions.
AuthorsSala C, Andreose JS, Fumagalli G, Lømo T
JournalJ Neurosci
PubMed ID7823160
The expression and content of calcitonin gene-related peptide (CGRP) and secretogranin II (SgII) in adult rat motor neurons were examined by in situ hybridization, Northern blot analysis, and immunocytochemistry. Normal motor nerve terminals did not contain detectable CGRP or SgII. Ten to 15 days after a peripheral nerve crush about ... More
Gbeta gamma mediate differentiation of vascular smooth muscle cells.
AuthorsReusch HP, Schaefer M, Plum C, Schultz G, Paul M
JournalJ Biol Chem
PubMed ID11279222
Proliferation and subsequent dedifferentiation of vascular smooth muscle (VSM) cells contribute to the pathogenesis of atherosclerosis and postangioplastic restenosis. The dedifferentiation of VSM cells in vivo or in cell culture is characterized by a loss of contractile proteins such as smooth muscle-specific alpha-actin and myosin heavy chain (SM-MHC). Serum increased ... More
Vascular endothelial growth factor (VEGF)-driven actin-based motility is mediated by VEGFR2 and requires concerted activation of stress-activated protein kinase 2 (SAPK2/p38) and geldanamycin-sensitive phosphorylation of focal adhesion kinase.
In endothelial cells, vascular endothelial growth factor (VEGF) induces an accumulation of stress fibers associated with new actin polymerization and rapid formation of focal adhesions at the ventral surface of the cells. This cytoskeletal reorganization results in an intense motogenic activity. Using porcine endothelial cells expressing one or the other ... More
Trafficking, assembly, and function of a connexin43-green fluorescent protein chimera in live mammalian cells.
AuthorsJordan K, Solan JL, Dominguez M, Sia M, Hand A, Lampe PD, Laird DW
JournalMol Biol Cell
PubMed ID10359613
To examine the trafficking, assembly, and turnover of connexin43 (Cx43) in living cells, we used an enhanced red-shifted mutant of green fluorescent protein (GFP) to construct a Cx43-GFP chimera. When cDNA encoding Cx43-GFP was transfected into communication-competent normal rat kidney cells, Cx43-negative Madin-Darby canine kidney (MDCK) cells, or communication-deficient Neuro2A ... More
Clathrin-mediated endocytosis and recycling of autocrine motility factor receptor to fibronectin fibrils is a limiting factor for NIH-3T3 cell motility.
AuthorsLe PU, Benlimame N, Lagana A, Raz A, Nabi IR
JournalJ Cell Sci
PubMed ID10954421
Autocrine motility factor receptor (AMF-R) is internalized via a clathrin-independent pathway to smooth endoplasmic reticulum tubules. This endocytic pathway is shown here to be inhibited by methyl-(beta)-cyclodextrin (m(beta)CD) implicating caveolae or caveolae-like structures in AMF internalization to smooth ER. AMF-R is also internalized via a clathrin-dependent pathway to a transferrin ... More
A double immunofluorescent method for simultaneous analysis of progesterone-dependent changes in proliferation and the oestrogen receptor in endometrium of rhesus monkeys.
AuthorsOkulicz WC, Balsamo M
JournalJ Reprod Fertil
PubMed ID8107038
A double immunofluorescent technique that permitted the nuclear localization of both the oestrogen receptor and the Ki-67 antigen (marker of cell proliferation) within the same tissue section was developed and used to determine the relationship between the oestrogen receptor and proliferation within the major zones of the primate endometrium. Endometrial ... More
Subcellular localization of CD80 receptors is dependent on an intact cytoplasmic tail and is required for CD28-dependent T cell costimulation.
AuthorsDoty RT, Clark EA
JournalJ Immunol
PubMed ID8871621
CD28 provides a major costimulatory signal to T cells when it is cross-linked with mAb, immobilized recombinant ligand (CD80Ig or CD86Ig), or ligand-bearing cells but not when it is bound by specific Fab fragments or monomeric ligand. We wanted to determine how monomeric CD80 could cross-link CD28 since CD80 is ... More
Polyethylene glycol-modified GM-CSF expands CD11b(high)CD11c(high) but notCD11b(low)CD11c(high) murine dendritic cells in vivo: a comparative analysis with Flt3 ligand.
AuthorsDaro E, Pulendran B, Brasel K, Teepe M, Pettit D, Lynch DH, Vremec D, Robb L, Shortman K, McKenna HJ, Maliszewski CR, Maraskovsky E
JournalJ Immunol
PubMed ID10861034
Dendritic cells (DC) are potent APCs that can be characterized in the murine spleen as CD11b(high)CD11c(high) or CD11b(low)CD11c(high). Daily injection of mice of Flt3 ligand (FL) into mice transiently expands both subsets of DC in vivo, but the effect of administration of GM-CSF on the expansion of DC in vivo ... More
Immunohistochemical signal amplification by catalyzed reporter deposition and its application in double immunostaining.
AuthorsHunyady B, Krempels K, Harta G, Mezey E
JournalJ Histochem Cytochem
PubMed ID8985127
The biotinyl-tyramide substrate of the horseradish peroxidase enzyme has been recently introduced to amplify immunohistochemical signals. We applied either fluorochromeor biotin-conjugated tyramine to improve the detection of different antigens in sections of rat stomach, pancreas, and hypothalamus. A ten- to 100-fold increase in staining efficiency was achieved, depending on the ... More
Molecular cloning, expression, and mapping of the high affinity actin-capping domain of chicken cardiac tensin.
AuthorsChuang JZ, Lin DC, Lin S
JournalJ Cell Biol
PubMed ID7896874
Tensin, an actin filament capping protein first purified from chicken gizzard, is localized to various types of adherens junctions in muscle and nonmuscle cells. In this paper, we describe the isolation and sequencing of tensin cDNA from a chicken cardiac library. The 6.3-kb chicken cardiac tensin cDNA encodes an open ... More
Triple immunofluorescence confocal laser scanning microscopy: spatial correlation of novel cellular differentiation markers in human muscle biopsies.
AuthorsSchubert W
JournalEur J Cell Biol
PubMed ID1718754
Two different methods for triple immunofluorescence imaging with a confocal laser scanning microscope (CLSM) are described. The methods enable spatial "mapping" of 3 different epitope distribution patterns simultaneously in one tissue section. The key to triple imaging includes: (a) specific immunolabeling with 3 different mouse monoclonal antibodies (mAbs), (b) localization ... More
The apical submembrane cytoskeleton participates in the organization of the apical pole in epithelial cells.
AuthorsSalas PJ, Rodriguez ML, Viciana AL, Vega-Salas DE, Hauri HP
JournalJ Cell Biol
PubMed ID9128248
In a previous publication (Rodriguez, M.L., M. Brignoni, and P.J.I. Salas. 1994. J. Cell Sci. 107: 3145-3151), we described the existence of a terminal web-like structure in nonbrush border cells, which comprises a specifically apical cytokeratin, presumably cytokeratin 19. In the present study we confirmed the apical distribution of cytokeratin ... More
Immunohistochemical distribution of the prohormone convertase PC5-A in rat brain.
AuthorsVilleneuve P, Seidah NG, Beaudet A
JournalNeuroscience
PubMed ID10408612
Prohormone convertase 5 is an endoprotease of the kexin/subtilisin-like family, which has been postulated to play a role in the proteolytic maturation of a variety of pro-peptides in the mammalian brain. In order to gain insight into the functional role of prohormone convertase 5 in the central nervous system, the ... More
Differential binding of the lectins Griffonia simplicifolia I and Lycopersicon esculentum to microvascular endothelium: organ-specific localization and partial glycoprotein characterization.
AuthorsPorter GA, Palade GE, Milici AJ
JournalEur J Cell Biol
PubMed ID2328740
The lectins Griffonia simplicifolia I and Lycopersicon esculentum were used to assess the presence of endothelium-specific glycoproteins in the microvasculature of the rat myocardium, diaphragm and superficial cerebral cortex. Organs fixed by intravascular perfusion were processed to obtain semithin (0.5 micron) and thin (less than 0.1 micron) frozen sections that ... More
Physiological, morphological, and histochemical characterization of three classes of interneurons in rat neostriatum.
AuthorsKawaguchi Y
JournalJ Neurosci
PubMed ID7693897
Interneurons in lateral part of neostriatum were studied in isolated slices from juvenile rats (16-20 d postnatal) by whole-cell, current-clamp recording at 33-34 degrees C, followed by intracellular staining with biocytin and double immunocytochemical or histochemical staining for parvalbumin, ChAT, and NADPH diaphorase. Medium-sized spiny neurons (MS cells) had distal ... More
The export of major histocompatibility complex class I molecules from the endoplasmic reticulum of rat brown adipose cells is acutely stimulated by insulin.
Major histocompatibility complex class I (MHC-I) molecules have been implicated in several nonimmunological functions including the regulation and intracellular trafficking of the insulin-responsive glucose transporter GLUT4. We have used confocal microscopy to compare the effects of insulin on the intracellular trafficking of MHC-I and GLUT4 in freshly isolated rat brown ... More
Segregation of the pathways leading to cortical reaction and cell cycle activation in the rat egg.
AuthorsRaz T, Ben-Yosef D, Shalgi R
JournalBiol Reprod
PubMed ID9472928
At fertilization of the mammalian egg, resumption of the cell cycle and the cortical reaction are two events of egg activation, correlated with an increase in intracellular Ca2+ concentration and activation of protein kinase C. To evaluate the pathways leading to both events, rat eggs were parthenogenetically activated by the ... More
Proteasome inhibitor-induced apoptosis of glioma cells involves the processing of multiple caspases and cytochrome c release.
AuthorsWagenknecht B, Hermisson M, Groscurth P, Liston P, Krammer PH, Weller M
JournalJ Neurochem
PubMed ID11080180
The proteasome is a multiprotein complex that is involved in the intracellular protein degradation in eukaryotes. Here, we show that human malignant glioma cells are susceptible to apoptotic cell death induced by the proteasome inhibitors, MG132 and lactacystin. The execution of the apoptotic death program involves the processing of caspases ... More
Kinetochore immunofluorescence in micronuclei: a rapid method for the in situ detection of aneuploidy and chromosome breakage in human fibroblasts.
AuthorsHennig UG, Rudd NL, Hoar DI
JournalMutat Res
PubMed ID3057373
We have developed a rapid and simple immunodetection assay for the in situ identification of aneuploidy in mitotic fibroblasts. Kinetochore (centromere)-containing micronuclei can be detected easily and rapidly by immunofluorescence. The action of colchicine and its derivatives on the mitotic spindle apparatus of mammalian cells induces chromosome lag and aneuploidy. ... More
Heterogeneous processing and zona pellucida binding activity of pig zonadhesin.
AuthorsHickox JR, Bi M, Hardy DM
JournalJ Biol Chem
PubMed ID11526117
Zonadhesin is a mosaic protein in sperm membrane fractions that binds directly and in a species-specific manner to the extracellular matrix (zona pellucida) of the oocyte. The active form of pig zonadhesin from capacitated, epididymal spermatozoa comprises two covalently associated polypeptide chains of M(r) 105,000 (p105) and M(r) 45,000 (p45). ... More
Preferential incorporation of 3'-azido-2',3'-dideoxythymidine into telomeric DNA and Z-DNA-containing regions of Chinese hamster ovary cells.
AuthorsOlivero OA, Poirier MC
JournalMol Carcinog
PubMed ID8397798
3'-Azido-2',3'-dideoxythymidine (azidothymidine; AZT) induces bone marrow toxicity in patients chronically given therapeutic doses of drug and is tumorigenic in rodents, inducing squamous cell tumors in vaginal tissues of mice and rats. In the study reported here, we explored the incorporation of AZT into specific regions of mammalian chromosomal DNA. CHO ... More
A strategy for multiple immunophenotyping by image cytometry: model studies using latex microbeads labeled with seven streptavidin-bound fluorochromes.
AuthorsGothot A, Grosdent JC, Paulus JM
JournalCytometry
PubMed ID8800554
Multiple immunophenotyping is aimed at identifying several cell populations in a single labeling procedure by their ability to bind combinations of specific labeled antibodies. The present work demonstrates the simultaneous discrimination by using image cytometry of aminomethylcoumarin acetate (AMCA), Lucifer yellow (LY), fluorescein isothiocyanate (FITC), R-phycoerythrin (PE), PE-Texas red tandem ... More
The cytosolic termini of the beta- and gamma-ENaC subunits are involved in the functional interactions between cystic fibrosis transmembrane conductance regulator and epithelial sodium channel.
AuthorsJi HL, Chalfant ML, Jovov B, Lockhart JP, Parker SB, Fuller CM, Stanton BA, Benos DJ
JournalJ Biol Chem
PubMed ID10821834
Epithelial sodium channel (ENaC) and cystic fibrosis transmembrane conductance regulator (CFTR) are co-localized in the apical membrane of many epithelia. These channels are essential for electrolyte and water secretion and/or reabsorption. In cystic fibrosis airway epithelia, a hyperactivated epithelial Na(+) conductance operates in parallel with defective Cl(-) secretion. Several groups ... More
NH2-Terminal targeting motifs direct dual specificity A-kinase-anchoring protein 1 (D-AKAP1) to either mitochondria or endoplasmic reticulum.
AuthorsHuang LJ, Wang L, Ma Y, Durick K, Perkins G, Deerinck TJ, Ellisman MH, Taylor SS
JournalJ Cell Biol
PubMed ID10352013
Subcellular localization directed by specific targeting motifs is an emerging theme for regulating signal transduction pathways. For cAMP-dependent protein kinase (PKA), this is achieved primarily by its association with A-kinase-anchoring proteins (AKAPs). Dual specificity AKAP1, (D-AKAP1) binds to both type I and type II regulatory subunits and has two NH2-terminal ... More
Dynamic, electronically switchable surfaces for membrane protein microarrays.
AuthorsTang CS, Dusseiller M, Makohliso S, Heuschkel M, Sharma S, Keller B, Vörös J
JournalAnal Chem
PubMed ID16448043
Microarray technology is a powerful tool that provides a high throughput of bioanalytical information within a single experiment. These miniaturized and parallelized binding assays are highly sensitive and have found widespread popularity especially during the genomic era. However, as drug diagnostics studies are often targeted at membrane proteins, the current ... More
Immunolabeling reveals cellular localization of the N-methyl-D-aspartate receptor subunit NR2B in neurosecretory cells but not astrocytes of the rat magnocellular nuclei.
Previous studies suggest that activation of N-methyl-D-aspartate (NMDA) receptors facilitates phasic firing and spike clustering displayed by magnocellular neuroendocrine cells (MNCs) of the supraoptic (SON) and paraventricular nucleus of the hypothalamus (PVN). Osmotic stimulation produces similar activity patterns which, in turn, can lead to enhanced release of vasopressin and oxytocin ... More
Immunofluorescent analysis of estrogen induction of progesterone receptor in the rhesus uterus.
AuthorsOkulicz WC, Savasta AM, Hoberg LM, Longcope C
JournalEndocrinology
PubMed ID2666107
Estrogen (E) has been shown to induce an increase in progesterone (P) receptor (PR) concentration in uterine tissue of both rodents and primates. Because of the presence of different cell types within the uterus, we were interested in determining whether estrogen-induced PR were cell type specific in the nonhuman primate ... More
Differential use of myristoyl groups on neuronal calcium sensor proteins as a determinant of spatio-temporal aspects of Ca2+ signal transduction.
Authors O'Callaghan Dermott W; Ivings Lenka; Weiss Jamie L; Ashby Michael C; Tepikin Alexei V; Burgoyne Robert D;
JournalJ Biol Chem
PubMed ID11836243
The localizations of three members of the neuronal calcium sensor (NCS) family were studied in HeLa cells. Using hippocalcin-EYFP and NCS-1-ECFP, it was found that their localization differed dramatically in resting cells. NCS-1 had a distinct predominantly perinuclear localization (similar to trans-Golgi markers), whereas hippocalcin was present diffusely throughout the ... More