| RRC ID |
33960
|
| Author |
Davis WP, Janssen YM, Mossman BT, Taatjes DJ.
|
| Title |
Simultaneous triple fluorescence detection of mRNA localization, nuclear DNA, and apoptosis in cultured cells using confocal scanning laser microscopy.
|
| Journal |
Histochem Cell Biol
|
| Abstract |
We describe a multifluorescence labeling technique for simultaneous detection of mRNA, nuclear DNA, and apoptosis in cultured cells. Digoxigenin-labeled cRNA probes were used to study proto-oncogene expression in rat pleural mesothelial cells undergoing apoptosis following exposure to crocidolite asbestos or hydrogen peroxide (H2O2). Hybridized cRNA probe was detected by immunolocalization with an anti-digoxigenin monoclonal primary and fluorophore-conjugated anti-mouse secondary antibody. Cells undergoing apoptosis were simultaneously identified by the TdT-mediated biotin-dUTP nick-end labeling (TUNEL) method and a streptavidin-conjugated far-red fluorophore, and nuclear DNA was stained with oxazole yellow dimer (YOYO-1). With confocal scanning laser microscopy, we demonstrated increased c-jun mRNA expression within the cytoplasm of both TUNEL-positive and non-apoptotic cells following exposure to either crocidolite asbestos or H2O2. Thus, this technique represents a useful in vivo approach for evaluating apoptosis-associated gene expression with confocal scanning laser microscopy.
|
| Volume |
108(4-5)
|
| Pages |
307-11
|
| Published |
1997-1-1
|
| DOI |
10.1007/s004180050170
|
| PMID |
9387922
|
| MeSH |
Animals
Apoptosis*
Asbestos, Crocidolite
Benzoxazoles / chemistry
Carbocyanines / chemistry
Cell Nucleus / chemistry
Cells, Cultured
DNA / analysis*
Fluorescence
Fluorescent Dyes / chemistry
In Situ Hybridization, Fluorescence / methods*
Microscopy, Confocal / methods*
Proto-Oncogene Proteins c-fos / genetics
Proto-Oncogene Proteins c-jun / genetics
Quinolinium Compounds / chemistry
RNA, Messenger / analysis*
Rats
Ribonucleases / metabolism
Water
|
| IF |
3.418
|
| Times Cited |
15
|
|
WOS Category
|
MICROSCOPY
CELL BIOLOGY
|
| Resource |
| DNA material |
pBS rcjun-1 (RDB01130) |
