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Detection of Senescence Markers During Mammalian Embryonic Development

Protocol
First Online:
Part of the Methods in Molecular Biology book series (MIMB, volume 1534)

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 
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Notes

Acknowledgments

This work was funded by Grants SAF2010-18829 and SAF2013-49082-P to W.M.K. from the Spanish Ministry for Economy and Competitiveness, the Agència de Gestió d’Ajuts Universitaris i de Recerca (AGAUR) from the Generalitat de Catalunya, and CRG core funding.

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Copyright information

© Springer Science+Business Media New York 2017

Authors and Affiliations

  1. 1.Centre for Genomic Regulation (CRG)The Barcelona Institute of Science and TechnologyBarcelonaSpain
  2. 2.Universitat Pompeu Fabra (UPF)BarcelonaSpain
  3. 3.Program in Neurosciences and Mental HealthHospital for Sick ChildrenTorontoCanada

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