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

doi:10.1083/jcb.119.3.493

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

Y Gavrieli, Y Sherman and SA Ben-Sasson
Department of Experimental Medicine, Hebrew University-Hadassah Medical School, Jerusalem, Israel.

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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Okunaga, T., Urata, Y., Goto, S., Matsuo, T., Mizota, S., Tsutsumi, K., Nagata, I., Kondo, T., Ihara, Y. (2006). Calreticulin, a Molecular Chaperone in the Endoplasmic Reticulum, Modulates Radiosensitivity of Human Glioblastoma U251MG Cells.. Cancer Res. 66: 8662-8671 [Abstract] [Full Text]
Homma, S., Iwasaki, M., Shelton, G. D., Engvall, E., Reed, J. C., Takayama, S. (2006). BAG3 Deficiency Results in Fulminant Myopathy and Early Lethality. Am. J. Pathol. 169: 761-773 [Abstract] [Full Text]
Sarda-Mantel, L., Coutard, M., Rouzet, F., Raguin, O., Vrigneaud, J.-M., Hervatin, F., Martet, G., Touat, Z., Merlet, P., Le Guludec, D., Michel, J.-B. (2006). 99mTc-Annexin-V Functional Imaging of Luminal Thrombus Activity in Abdominal Aortic Aneurysms. Arterioscler. Thromb. Vasc. Bio. 26: 2153-2159 [Abstract] [Full Text]
Geiger, G. A., Parker, S. E., Beothy, A. P., Tucker, J. A., Mullins, M. C., Kao, G. D. (2006). Zebrafish as a "Biosensor"? Effects of Ionizing Radiation and Amifostine on Embryonic Viability and Development. Cancer Res. 66: 8172-8181 [Abstract] [Full Text]
Marshall, J. R., Sakr, W., Wood, D., Berry, D., Tangen, C., Parker, F., Thompson, I., Lippman, S. M., Lieberman, R., Alberts, D., Jarrard, D., Coltman, C., Greenwald, P., Minasian, L., Crawford, E. D. (2006). Design and Progress of a Trial of Selenium to Prevent Prostate Cancer among Men with High-Grade Prostatic Intraepithelial Neoplasia.. Cancer Epidemiol. Biomarkers Prev. 15: 1479-1484 [Abstract] [Full Text]
Herzog, E. L., Van Arnam, J., Hu, B., Krause, D. S. (2006). Threshold of lung injury required for the appearance of marrow-derived lung epithelia.. Stem Cells 24: 1986-1992 [Abstract] [Full Text]
Kinoshita, M., McDannold, N., Jolesz, F. A., Hynynen, K. (2006). Noninvasive localized delivery of Herceptin to the mouse brain by MRI-guided focused ultrasound-induced blood-brain barrier disruption. Proc. Natl. Acad. Sci. USA 103: 11719-11723 [Abstract] [Full Text]
Sappington, R. M., Chan, M., Calkins, D. J. (2006). Interleukin-6 protects retinal ganglion cells from pressure-induced death.. IOVS 47: 2932-2942 [Abstract] [Full Text]
Laabich, A., Vissvesvaran, G. P., Lieu, K. L., Murata, K., McGinn, T. E., Manmoto, C. C., Sinclair, J. R., Karliga, I., Leung, D. W., Fawzi, A., Kubota, R. (2006). Protective Effect of Crocin against Blue Light- and White Light-Mediated Photoreceptor Cell Death in Bovine and Primate Retinal Primary Cell Culture.. IOVS 47: 3156-3163 [Abstract] [Full Text]
Osborne, N. N., Wood, J. P. M. (2006). The {beta}-Adrenergic Receptor Antagonist Metipranolol Blunts Zinc-Induced Photoreceptor and RPE Apoptosis.. IOVS 47: 3178-3186 [Abstract] [Full Text]
Dupont-Versteegden, E. E., Fluckey, J. D., Knox, M., Gaddy, D., Peterson, C. A. (2006). Effect of flywheel-based resistance exercise on processes contributing to muscle atrophy during unloading in adult rats. J. Appl. Physiol. 101: 202-212 [Abstract] [Full Text]