In situ immunodetection of activated caspase-3 in apoptotic neurons in the developing nervous system (original) (raw)

Cell Death & Differentiation volume 5, pages 1004–1016 (1998)Cite this article

Abstract

Activation of caspase-3 requires proteolytic processing of the inactive zymogen into p18 and p12 subunits. We generated a rabbit polyclonal antiserum, CM1, which recognizes the p18 subunit of cleaved caspase-3 but not the zymogen. CM1 demonstrated an apparent specificity for activated caspase-3 by specifically immunolabeling only apoptotic but not necrotic cortical neurons in vitro. In the embryonic mouse nervous system, CM1 immunoreactivity was detected in neurons undergoing programmed cell death and was markedly increased in Bcl-xL-deficient embryos and decreased in Bax-deficient embryos. CM1 immunoreactivity was absent in the nervous system of caspase-3-deficient mouse embryos and in neurons cultured from caspase-3-deficient mice. Along with neuronal somata, extensive neuritic staining was seen in apoptotic neurons. These studies indicate that caspase-3 is activated during apoptosis in the developing nervous system in vivo and that CM1 is a useful reagent for its in situ detection.

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  1. Anu Srinivasan and Kevin A Roth: A Srinivasan and KA Roth contributed equally to this work

Authors and Affiliations

  1. IDUN Pharmaceuticals, Inc., La Jolla, California, USA
    Anu Srinivasan, Robert O Sayers, Angela M Wong, Lawrence C Fritz & Kevin J Tomaselli
  2. Department of Pathology, Washington University School of Medicine, St. Louis, Missouri, USA
    Kevin A Roth & Kenneth S Shindler

Authors

  1. Anu Srinivasan
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  2. Kevin A Roth
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  3. Robert O Sayers
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  4. Kenneth S Shindler
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  5. Angela M Wong
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  6. Lawrence C Fritz
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  7. Kevin J Tomaselli
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Correspondence toAnu Srinivasan or Kevin J Tomaselli.

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Srinivasan, A., Roth, K., Sayers, R. et al. In situ immunodetection of activated caspase-3 in apoptotic neurons in the developing nervous system.Cell Death Differ 5, 1004–1016 (1998). https://doi.org/10.1038/sj.cdd.4400449

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