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.

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

Distribution of TaqI sites along human chromosomes revealed by in situ enzyme-nick translation

Genome ◽  
1993 ◽  
Vol 36 (1) ◽  
pp. 202-205 ◽  
Author(s):  
I. Tagarro ◽  
J. J. Gonzalez-Aguilera ◽  
A. M. Fernandez-Peralta ◽  
G. F. de Stefano ◽  
L. Ferrucci
Keyword(s):  
Complete Information ◽  
Human Chromosomes ◽  
Nick Translation ◽  
In Situ Nick Translation ◽  
Translation Procedure

We have studied the relative richness of TaqI sites along human chromosomes by means of a nonradioactive in situ enzyme-nick translation procedure. Regions with a higher content of these sequences are shown to be the noncentromeric heterochromatin blocks, whereas within euchromatin, terminal R-bands are the domains more enriched in these sites. Results obtained suggest that the method of performing enzyme digestions using time as a variable, and then in situ nick translation, provides much more complete information about the distribution of enzyme sequences along chromosomes than standard enzyme digestions.Key words: TaqI, nick translation, biotin, T-bands, heterochromatin.

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

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 DNase I nick translation in situ on meiotic chromosomes of the mouse, Mus musculus

1988 ◽  
Vol 90 (4) ◽  
pp. 629-634
Author(s):  
R. Raman ◽  
A.P. Singh ◽  
I. Nanda
Keyword(s):  
Sex Chromosomes ◽  
Mus Musculus ◽  
Meiotic Prophase ◽  
Cell Types ◽  
Dnase I ◽  
Translation Method ◽  
Nick Translation ◽  
Meiotic Chromosomes

DNase-I-sensitive sites have been located on the meiotic chromosomes of the mouse, Mus musculus, by the in situ DNase I nick-translation method. We find that: (1) of all the cell types studied, pachytene nuclei are the most sensitive to DNase I; (2) in diplotene the nicks occur preferentially in the vicinity of chiasmata; (3) the sex chromosomes are also sensitive to the enzyme despite their transcriptional quiescence; and (4) in the sex bivalent the nicks are primarily observed in the putative region of recombination. We conclude that, in addition to discriminating between the transcriptionally active and inactive states of chromatin, DNase I identifies recombination-specific chromatin changes in meiotic prophase.

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