Recent developments in the detection of deoxyribonucleic acid synthesis by 33258 Hoechst fluorescence.

Latt, Samuel A.; Stetten, Gail; Juergens, Lois A.; Willard, Huntington F.; Scher, Charles D.

doi:10.1177/23.7.1095650

Cited by 146 publications

(67 citation statements)

References 29 publications

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“…When mouse cells are grown in BrdU for one replication cycle and stained with the fluorescent dyes Hoechst 33258 or DAPI, an unequal brightness between sister chromatics is observed in the centromeric regions of mitotic chromosomes [9,10]. Similar observations were made with human cells, but fewer chromosomes were affected [11][12][13][14][15]. Reports of this phenomenon, which was given the name "lateral asymmetry," were fiist published over 20 years ago along with a plausible mechanistic explanation.…”

Section: Scientific Approach and Accomplishmentssupporting

confidence: 53%

Applications of Strand-Specific in situ Hybridization

Goodwin¹,

Meyne²,

Bailey³

et al. 1997

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Los AfenrosNational Mmratory, an affirmativeacfbnkqual opportunityarnpfoyer,is oparsted by the Unfveraityof California for the U.S. Daparfmeti of Energy under contractW-7405-ENG-3S. By aoce@ance of thb articfe, the pubtbher recognizesthat the U.S. Governrnent retains a nonexcfuak'e,myaftytree licenseto pubibfr or reproduoethe pubtbhad forrrrof this oonfrtbufbn,or to SUOW others to do SO,for U.S. Governmentpwpoees. Los AfamoaNational Laboratoryrequ@.s that the publleher Mer#lfy thb artfcfeas work petforrnedunder the auspkas of the U.S. Departrnari of Energy. Los Afsmoa Nafbnaf I_rboratoryatmrrgtysupportsacademic freedom and a researdw's rightto pWietV as an InafiWtbn, fmwever, the Laboratorydoes not andoma the vfewpoinfof a publkafbn or guarantee fts tecfmkal comecfneee.FarrI&3s ( Reseamh and Development (LDRD) project at the Los Alamos National Laboratory (LANL). Fluorescence in situ hybridization (FISH) is used to determine the location of specif3c DNA sequences on chromosomes. It is an effective tool in genomic mapping and is finding increasing use in medical diagnosis. A "strand-specKlc" version of FISH has been developed in the Life Sciences Division of LANL. The new procedure, named CO-FISH, reveals not only location but also the 5'-to-3' direction of a target sequence, such as the sense strand of a gene. This project was designed to investigate applications of the new technique. Strandspedlc FISH was found to be useful and informative for genomic mapping of repetitive DNA sequences. The method provide a valuable new tool for investigating the mechanisms of aneuploidy inducing agents and the cytogenetic phenomena called lateral asymmetry. Finally, using strand-specific FISH, we we~able to detect certain types of chromosome aberrations (isochromosomes, inversions and Robertsonian translocations) that can be difficult to observe with standard techniques. Background and Research Objectives

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Section: Scientific Approach and Accomplishmentssupporting

confidence: 53%

Applications of Strand-Specific in situ Hybridization

Goodwin¹,

Meyne²,

Bailey³

et al. 1997

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“…For nuclear staining, immunostained wing discs were incubated for 15 minutes at room temperature in a 1:1000 dilution of Hoechst 33258 (Invitrogen) (Latt et al, 1975) in PBS-T (PBS with 0.3% Triton X-100), followed by two 10-minute washes in PBS-T prior to mounting under coverslips.…”

Section: Immunohistologymentioning

confidence: 99%

Protein trafficking abnormalities inDrosophilatissues with impaired activity of the ZIP7 zinc transporter Catsup

et al. 2013

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Developmental patterning requires the precise interplay of numerous intercellular signaling pathways to ensure that cells are properly specified during tissue formation and organogenesis. The spatiotemporal function of the Notch signaling pathway is strongly influenced by the biosynthesis and intracellular trafficking of signaling components. Receptors and ligands must be trafficked to the cell surface where they interact, and their subsequent endocytic internalization and endosomal trafficking is crucial for both signal propagation and its down-modulation. In a forward genetic screen for mutations that alter intracellular Notch receptor trafficking in Drosophila epithelial tissues, we recovered mutations that disrupt the Catsup gene, which encodes the Drosophila ortholog of the mammalian ZIP7 zinc transporter. Loss of Catsup function causes Notch to accumulate abnormally in the endoplasmic reticulum (ER) and Golgi compartments, resulting in impaired Notch signaling. In addition, Catsup mutant cells exhibit elevated ER stress, suggesting that impaired zinc homeostasis causes increased levels of misfolded proteins within the secretory compartment.

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“…In contrast, Hoechst 3342 is a blue fluorescent dye used to stain DNA and allows the differentiation of single cells in microscopy samples (Fig. 2E) (66). No significant variation in cell diameter was found between the wild type and the ⌬amy and ⌬lac mutants.…”

Section: Resultsmentioning

confidence: 85%

Reconstruction of mreB Expression in Staphylococcus aureus via a Collection of New Integrative Plasmids

Yepes

Koch

Waldvogel

et al. 2014

Appl Environ Microbiol

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Protein localization has been traditionally explored in unicellular organisms, whose ease of genetic manipulation facilitates molecular characterization. The two rod-shaped bacterial models Escherichia coli and Bacillus subtilis have been prominently used for this purpose and have displaced other bacteria whose challenges for genetic manipulation have complicated any study of cell biology. Among these bacteria is the spherical pathogenic bacterium Staphylococcus aureus. In this report, we present a new molecular toolbox that facilitates gene deletion in staphylococci in a 1-step recombination process and additional vectors that facilitate the insertion of diverse reporter fusions into newly identified neutral loci of the S. aureus chromosome. Insertion of the reporters does not add any antibiotic resistance genes to the chromosomes of the resultant strains, thereby making them amenable for further genetic manipulations. We used this toolbox to reconstitute the expression of mreB in S. aureus, a gene that encodes an actin-like cytoskeletal protein which is absent in coccal cells and is presumably lost during the course of speciation. We observed that in S. aureus, MreB is organized in discrete structures in association with the membrane, leading to an unusual redistribution of the cell wall material. The production of MreB also caused cell enlargement, but it did not revert staphylococcal shape. We present interactions of MreB with key staphylococcal cell wall-related proteins. This work facilitates the use S. aureus as a model system in exploring diverse aspects of cellular microbiology. A significant number of studies of protein localization has been traditionally performed by using bacterial models, which are relatively simple and tractable haploid organisms (1-3). However, only a few bacterial species, such as the model organisms Escherichia coli and Bacillus subtilis, are generally considered for these studies, mostly because of the availability of a large number of molecular tools. In an attempt to have a broader and more general idea of the fundamental processes within the microbial world, there is growing interest in exploring other bacterial species that differ in shape, developmental properties, or infective potential from conventional bacterial models (4-6). Among these species is the spherical bacterium Staphylococcus aureus. It is an opportunistic pathogen that causes hard-to-treat acute and chronic infections in humans (7). It is indeed the study of its pathogenic attributes that makes this bacterium very attractive. Besides the interest in S. aureus as a major human pathogen, it is also an appealing model to explore questions related to cell shape, cell division, and cell proliferation, because it propagates very efficiently under laboratory conditions and is morphologically different from the conventional models E. coli and B. subtilis.In order to genetically manipulate S. aureus in the laboratory, several batteries of plasmids have been constructed over the past years (8-12), yet performance of p...

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Recent developments in the detection of deoxyribonucleic acid synthesis by 33258 Hoechst fluorescence.

Cited by 146 publications

References 29 publications

Applications of Strand-Specific in situ Hybridization

Applications of Strand-Specific in situ Hybridization

Protein trafficking abnormalities inDrosophilatissues with impaired activity of the ZIP7 zinc transporter Catsup

Reconstruction of mreB Expression in Staphylococcus aureus via a Collection of New Integrative Plasmids

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