Detection of Senescence Markers During Mammalian Embryonic Development.
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.
Centre for Genomic Regulation (CRG), The Barcelona Institute of Science and Technology, Carrer del Dr. Aiguader 88, Barcelona, 08003, Spain.|Universitat Pompeu Fabra (UPF), Barcelona, Spain.|Program in Neurosciences and Mental Health, Hospital for Sick Children, 686 Bay Street, Toronto, ON, Canada, M5G 0A4.
Mekayla
M
Storer
Centre for Genomic Regulation (CRG), The Barcelona Institute of Science and Technology, Carrer del Dr. Aiguader 88, Barcelona, 08003, Spain. bill.keyes@crg.eu.|Universitat Pompeu Fabra (UPF), Barcelona, Spain. bill.keyes@crg.eu.
William M
WM
Keyes
D000818:Animals; D015415:Biomarkers; D016922:Cellular Senescence / Q000235:genetics; D050759:Cyclin-Dependent Kinase Inhibitor p21 / Q000235:genetics / Q000378:metabolism; D047108:Embryonic Development / Q000235:genetics; D006801:Humans; D007150:Immunohistochemistry / Q000379:methods; D008322:Mammals; D051379:Mice; D001616:beta-Galactosidase / Q000378:metabolism
D015415:Biomarkers; D050759:Cyclin-Dependent Kinase Inhibitor p21; D001616:beta-Galactosidase
Apoptosis; Biomarker; Cellular senescence; Development; Embryo; Limb; Neural tube; SAβ-gal; Whole-mount staining; p21