scholarly journals In situ Detection of Specific DNA Double Strand Breaks using Rolling Circle Amplification

Cell Cycle ◽  
2005 ◽  
Vol 4 (12) ◽  
pp. 1767-1773 ◽  
Author(s):  
Jia Li ◽  
C.S.H. Young ◽  
Paul M. Lizardia ◽  
David F. Stern
Keyword(s):  
Rolling Circle Amplification ◽  
Double Strand Breaks ◽  
Dna Double Strand Breaks ◽  
Rolling Circle ◽  
Strand Breaks ◽  
In Situ Detection

In Situ Detection of Messenger RNA Using Digoxigenin-Labeled Oligonucleotides and Rolling Circle Amplification

2001 ◽  
Vol 70 (3) ◽  
pp. 281-288 ◽  
Author(s):  
Yi Zhou ◽  
Margaret Calciano ◽  
Stefan Hamann ◽  
J.H. Leamon ◽  
Tod Strugnell ◽  
...  
Keyword(s):  
Messenger Rna ◽  
Rolling Circle Amplification ◽  
Rolling Circle ◽  
In Situ Detection

scholarly journals Transient Global Ischemia Triggers Expression of the DNA Damage-Inducible Gene GADD45 in the Rat Brain

1998 ◽  
Vol 18 (6) ◽  
pp. 646-657 ◽  
Author(s):  
Jun Chen ◽  
Koichi Uchimura ◽  
R. Anne Stetler ◽  
Raymond L. Zhu ◽  
Masaki Nakayama ◽  
...  
Keyword(s):  
Dna Damage ◽  
Cell Death ◽  
Blot Analysis ◽  
Double Strand Breaks ◽  
Dna Double Strand Breaks ◽  
Transient Ischemia ◽  
Strand Breaks ◽  
Transient Global Ischemia ◽  
Inducible Gene

Using in situ hybridization, Northern blot analysis, Western blot analysis, and immunocytochemistry, mRNA and protein expression of the novel DNA damage-inducible gene GADD45 was examined in the rat brain at 0.5, 2, 4, 8, 16, 24, 48, and 72 hours after 15 minutes of transient global ischemia. Transient ischemia produced by the four-vessel occlusion method resulted in DNA double-strand breaks and delayed neuronal cell death in vulnerable neurons of the hippocampal CA1 sector, the hilus, dorsal caudate-putamen, and thalamus, as shown by in situ DNA nick end-labeling and histologic staining. GADD45 mRNA was transiently increased in less-vulnerable regions such as the parietal cortex (up to 8 hours after ischemia) and dentate granule cells (up to 24 hours after ischemia) but was persistently increased in vulnerable neurons such as CA1 pyramidal neurons (up to 48 hours). GADD45 immunoreactivity was increased in both vulnerable and less-vulnerable regions at earlier reperfusion periods (4 to 16 hours), but thereafter immunoreactivity was decreased below control levels in most vulnerable regions before delayed cell death and DNA double-strand breaks. At 72 hours after transient ischemia, a moderate increase in GADD45 immunoreactivity was still detectable in some CA3 neurons and in a few surviving neurons in the CA1 region. Double staining performed at 16 to 72 hours after ischemia revealed that GADD45 immunoreactivity was persistently increased in neurons that did not develop DNA damage. Because GADD45 protein may participate in the DNA excision repair process and because it has been shown that this protein is also overexpressed in neurons that survive focal ischemia and kainate-induced epileptic seizures, the results reported here support the hypothesis that GADD45 could have a protective role in neuronal injury.

scholarly journals Detection of Low-Copy-Number Genomic DNA Sequences in Individual Bacterial Cells by Using Peptide Nucleic Acid-Assisted Rolling-Circle Amplification and Fluorescence In Situ Hybridization

2007 ◽  
Vol 73 (7) ◽  
pp. 2324-2328 ◽  
Author(s):  
Irina Smolina ◽  
Charles Lee ◽  
Maxim Frank-Kamenetskii
Keyword(s):  
Dna Sequences ◽  
Peptide Nucleic Acid ◽  
Rolling Circle Amplification ◽  
Bacterial Genome ◽  
Bacterial Cells ◽  
Rolling Circle ◽  
Low Copy Number ◽  
Short Signature ◽  
In Situ Detection

ABSTRACT An approach is proposed for in situ detection of short signature DNA sequences present in single copies per bacterial genome. The site is locally opened by peptide nucleic acids, and a circular oligonucleotide is assembled. The amplicon generated by rolling circle amplification is detected by hybridization with fluorescently labeled decorator probes.

Induction of in situ DNA double-strand breaks and apoptosis by 200 MeV protons and 10 MV X-rays in human tumour cell lines

2010 ◽  
Vol 87 (1) ◽  
pp. 57-70 ◽  
Author(s):  
Ariungerel Gerelchuluun ◽  
Zhengshan Hong ◽  
Lue Sun ◽  
Kenshi Suzuki ◽  
Toshiyuki Terunuma ◽  
...  
Keyword(s):  
Tumour Cell ◽  
Double Strand Breaks ◽  
X Rays ◽  
Dna Double Strand Breaks ◽  
Human Tumour ◽  
Strand Breaks ◽  
Tumour Cell Lines ◽  
Human Tumour Cell Lines ◽  
Mev Protons

scholarly journals A damaged genome’s transcriptional landscape through multilayered expression profiling around in situ-mapped DNA double-strand breaks

2017 ◽  
Vol 8 (1) ◽  
Author(s):  
Fabio Iannelli ◽  
Alessandro Galbiati ◽  
Ilaria Capozzo ◽  
Quan Nguyen ◽  
Brian Magnuson ◽  
...  
Keyword(s):  
Expression Profiling ◽  
Double Strand Breaks ◽  
Dna Double Strand Breaks ◽  
Strand Breaks ◽  
Transcriptional Landscape

scholarly journals Cell Type–Specific Quantification of Telomere Length and DNA Double-strand Breaks in Individual Lung Cells by Fluorescence In Situ Hybridization and Fluorescent Immunohistochemistry

2018 ◽  
Vol 66 (7) ◽  
pp. 485-495 ◽  
Author(s):  
Aernoud A. van Batenburg ◽  
Karin M. Kazemier ◽  
Ton Peeters ◽  
Matthijs F. M. van Oosterhout ◽  
Joanne J. van der Vis ◽  
...  
Keyword(s):  
Smooth Muscle ◽  
In Situ Hybridization ◽  
Fluorescence In Situ Hybridization ◽  
Telomere Length ◽  
Dna Sequences ◽  
Double Strand Breaks ◽  
Telomere Maintenance ◽  
Dna Double Strand Breaks ◽  
Strand Breaks

Telomeres are small repetitive DNA sequences at the ends of chromosomes which act as a buffer in age-dependent DNA shortening. Insufficient telomere repeats will be recognized as double-strand breaks. Presently, it is becoming more evident that telomere attrition, whether or not caused by mutations in telomere maintenance genes, plays an important role in many inflammatory and age-associated diseases. In this report, a method to (semi)quantitatively assess telomere length and DNA double-strand breaks in formalin-fixed paraffin-embedded (FFPE) tissue is described. Therefore, a novel combination of quantitative fluorescence in situ hybridization, tissue elution, and immunofluorescence staining techniques was developed. Caveolin-1 (type 1 pneumocytes), pro-surfactant protein C (type 2 pneumocytes), club cell-10 (club cells), and alpha smooth muscle actin (smooth muscle cells) markers were used to identify cell types. To visualize all the different probes, restaining the tissue by heat-mediated slide elution is essential. Fluorescent signals of telomeres and DNA double-strand breaks were quantified using the Telometer plugin of ImageJ. As example, we analyzed lung tissue from a familial pulmonary fibrosis patient with a mutation in the telomere-associated gene poly(A)-specific ribonuclease ( PARN). The protocol displays a novel opportunity to directly quantitatively link DNA double-strand breaks to telomere length in specific FFPE cells.

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