scholarly journals Tissue-specific DNA methylation in Haemanthus katharinae Bak. (Amaryllidaceae)

2014 ◽  
Vol 67 (2) ◽  
pp. 175-180 ◽  
Author(s):  
Tomasz Sakowicz ◽  
Maria J. Olszewska ◽  
Piotr Łuchniak ◽  
Joanna Kaźmierczak
Keyword(s):  
Dna Methylation ◽  
Root Meristem ◽  
Restriction Analysis ◽  
Blot Hybridization ◽  
Southern Blot Hybridization ◽  
Nick Translation ◽  
In Situ Nick Translation ◽  
Adult Leaf ◽  
Total Dna

The level of DNA methylation was compared in root meristem, adult leaf and endosperm of monocotyledonous species, <em>Haemanthus katharinae</em>, with the use of HPLC, restriction analysis, Southern blot hybridization and in situ nick-translation driven by restriction enzyme HhaI The highest level of 5-methylcytosine was observed in adult leaf whose nuclear chromatin is particularly condensed, and the lowest in endosperm. The level of DNA methylation of repetitive HaeIII 4006p sequence follows that of the total DNA.

scholarly journals Detection of DNA fragmentation in apoptosis: application of in situ nick translation to cell culture systems and tissue sections.

1993 ◽  
Vol 41 (7) ◽  
pp. 1023-1030 ◽  
Author(s):  
R Gold ◽  
M Schmied ◽  
G Rothe ◽  
H Zischler ◽  
H Breitschopf ◽  
...  
Keyword(s):  
Cell Death ◽  
Dna Fragmentation ◽  
Specific Cell ◽  
Nick Translation ◽  
Proliferating Cells ◽  
Cell Surface Antigens ◽  
Simultaneous Application ◽  
Tissue Sections ◽  
In Situ Nick Translation

Since DNA fragmentation is a key feature of programmed cell death (PCD) and also occurs in certain stages of necrosis, we have adapted the methodology of in situ nick-translation (ISNT) to detect DNA fragmentation on a single-cell level. We first established the technique for cell preparations. Apoptosis was induced by gamma-irradiation on freshly isolated rat thymocytes. After fixation procedures, ISNT was performed by overnight incubation either with fluorescein-12-dUTP or with digoxigenin-labeled 11-dUTP and DNA polymerase I. The enzymatic incorporation of labeled nucleotides at sites of DNA fragmentation was detected by flow cytometry either directly or indirectly with fluorescein-conjugated anti-digoxigenin. The quantitative results demonstrated close correlation with morphological essays for apoptosis, DNA gel electrophoresis, and ISNT. Proliferating cells determined by bromodeoxyuridine immunofluorescence were not labeled by ISNT. Immunocytochemistry for cell surface antigens in combination with ISNT allowed the identification of specific cell types undergoing PCD. Furthermore, the simultaneous application of photolabeling techniques with ethidium monoazide and ISNT led to the identification of DNA fragmentation in cells with still intact membranes. Extending ISNT to tissue sections of paraformaldehyde-fixed, paraffin-embedded material reliably revealed labeling of cells with typical morphological features of apoptosis. However, this technique was not useful in detecting early stages of necrotic cell death.

Microprocedure for in situ nick translation of chromosomes

1992 ◽  
Vol 62 (2) ◽  
pp. 150-153
Author(s):  
Li Zhengang ◽  
Howard L. Hosick ◽  
Kang Fan
Keyword(s):  
Nick Translation ◽  
In Situ Nick Translation

DNA single-strand breaks in postischemic gerbil brain detected by in situ nick translation procedure

1995 ◽  
Vol 200 (2) ◽  
pp. 129-132 ◽  
Author(s):  
Muneshige Tobita ◽  
Isao Nagano ◽  
Shozo Nakamura ◽  
Yasuto Itoyama ◽  
Kyuya Kogure
Keyword(s):  
Single Strand ◽  
Nick Translation ◽  
Strand Breaks ◽  
In Situ Nick Translation ◽  
Gerbil Brain ◽  
Single Strand Breaks ◽  
Translation Procedure

In situ nick translation of meiotic chromosomes to demonstrate homologous heterochromatin heterogeneity

Genome ◽  
1993 ◽  
Vol 36 (2) ◽  
pp. 268-270 ◽  
Author(s):  
J. de la Torre ◽  
C. López-Fernández ◽  
P. Herrero ◽  
J. Gosálvez
Keyword(s):  
Tandem Duplication ◽  
Dna Recombination ◽  
Nick Translation ◽  
Meiotic Chromosomes ◽  
In Situ Nick Translation ◽  
Living Species ◽  
Longitudinal Differentiation ◽  
Translation Procedure ◽  
Restriction Endonuclease Cleavage

The in situ nick translation procedure performed on fixed meiotic chromosomes partially cleaved with restriction endonucleases shows a different staining of homologous heterochromatic regions, which could be explained through a differential restriction endonuclease cleavage. Mutations occurring before massive tandem duplication and involving those DNA motifs that produce these heterochromatic blocks, together with the absence of DNA recombination that characterizes these particular regions, could explain the observed results. This method for chromosome labelling is most useful to demonstrate a certain level of heterochromatin heterogeneity that is present in the genome of living species but remained cryptic to other techniques that are also able to induce longitudinal differentiation of the chromosomes.Key words: cytogenetics, nick translation, meiosis, heterochromatin.

A new banding pattern of human chromosomes by in situ nick translation using ECO RI and biotin-dUTP

2008 ◽  
Vol 28 (2) ◽  
pp. 173-176 ◽  
Author(s):  
Jörn Bullerdiek ◽  
Jürgen Dittmer ◽  
Angelika Faehre ◽  
Sabine Bartnitzke ◽  
Volker Kasche ◽  
...  
Keyword(s):  
Banding Pattern ◽  
Human Chromosomes ◽  
Nick Translation ◽  
In Situ Nick Translation ◽  
Eco Ri

In situ nick-translation distinguishes between active and inactive X chromosomes

Nature ◽  
1983 ◽  
Vol 304 (5921) ◽  
pp. 88-90 ◽  
Author(s):  
Bat-Sheva Kerem ◽  
Ruth Goitein ◽  
Carmelit Richler ◽  
Menashe Marcus ◽  
Howard Cedar
Keyword(s):  
Nick Translation ◽  
X Chromosomes ◽  
In Situ Nick Translation

scholarly journals Influence of Mouse-Strain-Specific Factors on Position-Dependent Transgene DNA Methylation Patterns

1996 ◽  
Vol 45 (1-2) ◽  
pp. 243-244
Author(s):  
P.A. Koetsier ◽  
W. Doerfler
Keyword(s):  
Dna Methylation ◽  
Genetic Background ◽  
Promoter Activity ◽  
Blot Hybridization ◽  
Southern Blot Hybridization ◽  
Adenovirus Type ◽  
Methylation Pattern ◽  
Parent Of Origin ◽  
Methylation Patterns

In previous work from this laboratory, an inverse dependence was established for the adenovirus type 2 E2A late promoter between sequence-specific DNA methylation and promoter activity [1-5; for reviews see ref. 6, 7]. The effect of DNA methylation on promoter activity was also assessed in the transgenic mice, which were obtained from microinjections of unmethylated or in vitro HpaII-premethylated pAd2E2AL-CAT DNA [1] into F2 zygotes from B6D2F, (C57BL/6 × DBA/2) hybrid mice. In CAT assays carried out on organ extracts from the pAd2E2AL-CAT mice, the inverse relationship was confirmed [2].We studied the stability of the pAd2E2AL-CAT DNA methylation patterns in up to eight mouse generations and assessed the influence of the strain-specific genetic background. Three pAd2E2AL-CAT mouse lines were crossed with inbred DBA/2, C57BL/6 or B6D2F, mice. Parent-of-origin effects were controlled by exclusive hemizygous transgene transmission either via females or males. The founder animal of line 7-1 carried two groups of transgenes (A and B) on separate chromosomes. The transgene methylation patterns of the 7-1B transgenes and those of the lines 5-8 and 8-1 were stably transmitted.Southern blot hybridization experiments [8, 9] revealed that the 7-1A transgene methylation pattern was a cellular mosaic. In mixed-genetic-background offspring from 7-1A animals, 10% carried transgenes with HpaII-DNA methylation levels that were reduced from 40 to 10-15%. This finding suggested that in this background the factors that supported high methylation levels were dominant. When inbred DBA/2 mice were the mates, 40% of the siblings carried demethylated transgenes, whereas this ratio amounted to only 10% in C57BL/6 offspring (comparable to B6D2F1 crossings). Transgene methylation patterns were not detectably influenced by the parent-of-origin.

scholarly journals Papillomas and Carcinomas Associated with a Papillomavirus in European Harvest Mice (Micromys minutus)

1988 ◽  
Vol 25 (5) ◽  
pp. 356-361 ◽  
Author(s):  
J. P. Sundberg ◽  
M. K. O'Banion ◽  
A. Shima ◽  
C. Knupp ◽  
M. E. Reichmann
Keyword(s):  
Blot Hybridization ◽  
Southern Blot Hybridization ◽  
Skin Lesions ◽  
Sebaceous Carcinoma ◽  
Cutaneous Lesions ◽  
Pulmonary Tumors ◽  
Papillomavirus Dna ◽  
Inverted Papillomas ◽  
Micromys Minutus

Papillomaviruses, group-specific papillomavirus antigens, or extrachromosomal papillomavirus DNA were detected in cutaneous, mucocutaneous, and pulmonary tumors affecting a colony of European harvest mice (Micromys minutus). Skin lesions were classified as acanthomatous hyperplasia, epidermal inclusion cysts. squamous papillomas, inverted papillomas, trichoepitheliomas, and sebaceous carcinomas. Cutaneous horns (hyperkeratotic papillomas) were on mucocutaneous junctions of one animal. One mouse, with a cutaneous sebaceous carcinoma, had multiple pulmonary keratinaceous cysts. Papillomavirus antigens, detected by the avidin-biotin technique, were in 20 of 31 lesions tested. In contrast, by Southern blot hybridization all 28 lesions tested contained papillomavirus DNA. Papillomavirus DNA was demonstrated in two often benign cutaneous lesions by in situ hybridization.

scholarly journals Fatal Epstein-Barr virus-associated hemophagocytic syndrome

Blood ◽  
1993 ◽  
Vol 82 (11) ◽  
pp. 3259-3264 ◽  
Author(s):  
H Kikuta ◽  
Y Sakiyama ◽  
S Matsumoto ◽  
T Oh-Ishi ◽  
T Nakano ◽  
...  
Keyword(s):  
Epstein Barr Virus ◽  
Hemophagocytic Syndrome ◽  
Blot Hybridization ◽  
Southern Blot Hybridization ◽  
Barr Virus ◽  
Clonality Analysis ◽  
Serologic Evidence ◽  
Infected Cells ◽  
Epstein Barr

A virus-associated hemophagocytic syndrome is characterized by high fever, liver dysfunction, coagulation abnormalities, pancytopenia, and a benign histiocytic proliferation with prominent hemophagocytosis in bone marrow, lymph node, spleen, and liver. We describe six Japanese children with fatal Epstein-Barr virus (EBV)-associated hemophagocytic syndrome. Five of the six patients had serologic evidence of primary EBV infection at the onset of their diseases. EBV genomes were detected in all the patients by Southern blot hybridization or the polymerase chain reaction. Furthermore, clonality analysis of the EBV genome showed that EBV-infected cells proliferated monoclonally or biclonally in three examined patients. In situ hybridization study using EBV- encoded RNA 1 (EBER1) showed that EBER1 was detected in one of two examined liver tissues, which localized in hepatocytes.

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