Ultrastructural identification of apoptotic nuclei using the TUNEL technique.
Histochem J
; 28(4): 275-81, 1996 Apr.
Article
de En
| MEDLINE
| ID: mdl-8762059
ABSTRACT
We describe an ultrastructural adaptation of the method of terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labelling (TUNEL) for the identification of DNA fragmentation. Thin sections of tissue embedded in hydrophilic resin were nick end labelled with biotinylated dUTP which was subsequently labelled with avidin conjugated to gold particles. The technique was validated by labelling the nuclei of L929-8 cells treated with tumour necrosis factor alpha. These cells are known to respond to treatment with the factor by undergoing apoptosis. The method was then used on tissue from the chick embryo which is known to be undergoing programmed cell death. This tissue was from the neural tube and the posterior necrotic zone of the limb bud, where cells can be identified as undergoing apoptosis based on the morphology of their nuclei. The method specifically labelled heterochromatin adjacent to the nuclear envelope as well as that associated with the nucleolus of cells from regions of the embryo where programmed cell death was expected. In addition to labelling the nuclei of cells that were clearly undergoing apoptosis, the method also identified nuclei of apparently normal cells. This method, used in conjunction with corroborating techniques, provides a means for the early detection of cells undergoing DNA fragmentation, before the onset of gross apoptotic morphology, and in cells that do not show classical apoptotic characteristics.
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Collection:
01-internacional
Base de données:
MEDLINE
Sujet principal:
Microscopie électronique
/
Noyau de la cellule
/
Apoptose
Type d'étude:
Diagnostic_studies
/
Prognostic_studies
/
Screening_studies
Limites:
Animals
Langue:
En
Journal:
Histochem J
Année:
1996
Type de document:
Article
Pays d'affiliation:
Canada