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Oncogene-Induced Senescence

Volume 1534 of the series Methods in Molecular Biology pp 199-210

Date:

Detection of Senescence Markers During Mammalian Embryonic Development

  • Mekayla StorerAffiliated withCentre for Genomic Regulation (CRG), The Barcelona Institute of Science and TechnologyUniversitat Pompeu Fabra (UPF)Program in Neurosciences and Mental Health, Hospital for Sick Children
  • , William M. KeyesAffiliated withCentre for Genomic Regulation (CRG), The Barcelona Institute of Science and TechnologyUniversitat Pompeu Fabra (UPF) Email author 

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Abstract

Senescence-associated β-galactosidase (SAβ-gal) is a convenient histological technique used to identify senescent cells. Its ease of use is helpful to initially screen and detect senescent cells in heterogeneous cell populations both in vitro and in vivo. However, SAβ-gal staining is not an unequivocal marker of the senescent state, and diagnosis of such usually requires additional markers demonstrating an absence of proliferation and expression of cell-cycle inhibitors. Nonetheless, SAβ-gal remains one of the most widely used biomarkers of senescent cells. Recently, by measuring SAβ-gal activity, the expression of the cyclin-dependent kinase inhibitor p21 (waf1/cip1) and demonstrating a lack of proliferation, we identified senescent cells in the developing embryo. This chapter describes the methods for identifying cellular senescence in the embryo, detailing protocols for the detection of SAβ-gal activity in both sections and at the whole mount level, and immunohistochemistry protocols for the detection of additional biomarkers of senescence.

Key words

Cellular senescence SAβ-gal Whole-mount staining Embryo Biomarker Development Limb Neural tube p21 Apoptosis