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      Identification of programmed cell death in situ via specific labeling of nuclear DNA fragmentation

      The Journal of Cell Biology

      The Rockefeller University Press

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          Abstract

          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.

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          Author and article information

          Journal
          J Cell Biol
          J. Cell Biol.
          The Journal of Cell Biology
          The Rockefeller University Press
          0021-9525
          1540-8140
          1 November 1992
          : 119
          : 3
          : 493-501
          Article
          93016262
          2289665
          1400587
          Categories
          Articles

          Cell biology

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