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Identification of programmed cell death in situ via specific labeling of nuclear DNA fragmentation.
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Zeitschriftentitel: | The Journal of cell biology |
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Personen und Körperschaften: | , , |
In: | The Journal of cell biology, 119, 1992, 3, S. 493-501 |
Format: | E-Article |
Sprache: | Englisch |
veröffentlicht: |
Rockefeller University Press
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Schlagwörter: |
Zusammenfassung: | <jats:p>Programmed cell death (PCD) plays a key role in developmental biology and in maintenance of the steady state in continuously renewing tissues. Currently, its existence is inferred mainly from gel electrophoresis of a pooled DNA extract as PCD was shown to be associated with DNA fragmentation. Based on this observation, we describe here the development of a method for the in situ visualization of PCD at the single-cell level, while preserving tissue architecture. Conventional histological sections, pretreated with protease, were nick end labeled with biotinylated poly dU, introduced by terminal deoxy-transferase, and then stained using avidin-conjugated peroxidase. The reaction is specific, only nuclei located at positions where PCD is expected are stained. The initial screening includes: small and large intestine, epidermis, lymphoid tissues, ovary, and other organs. A detailed analysis revealed that the process is initiated at the nuclear periphery, it is relatively short (1-3 h from initiation to cell elimination) and that PCD appears in tissues in clusters. The extent of tissue-PCD revealed by this method is considerably greater than apoptosis detected by nuclear morphology, and thus opens the way for a variety of studies.</jats:p> |
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Umfang: | 493-501 |
ISSN: |
0021-9525
1540-8140 |
DOI: | 10.1083/jcb.119.3.493 |