PUBLICATION

Whole-mount Senescence-Associated Beta-Galactosidase (SA-β-GAL) Activity Detection Protocol for Adult Zebrafish

Authors
Marzullo, M., Maï, M.E., Ferreira, M.G.
ID
ZDB-PUB-220809-9
Date
2022
Source
Bio-protocol   12(13): (Journal)
Registered Authors
Ferreira, Miguel Godinho
Keywords
Aging, SA-β-GAL, Senescence, Whole organism, Zebrafish
MeSH Terms
none
PubMed
35937931 Full text @ Bio Protoc
Abstract
Senescence-associated beta-galactosidase (SA-β-GAL) is an enzyme that accumulates in the lysosomes of senescent cells, where it hydrolyses β-galactosides. With p16, it represents a well-recognized biomarker used to assess senescence both in vivo and in cell culture. The use of a chromogenic substrate, such as 5-bromo-4-chloro-3-indoyl-β-d-galactopyranoside (X-Gal), allows the detection of SA-β-GAL activity at pH 6.0 by the release of a visible blue product. Senescence occurs during aging and is part of the aging process itself. We have shown that prematurely aged zebrafish accumulate senescent cells detectable by SA-β-GAL staining in different tissues, including testis and gut. Here, we report a detailed protocol to perform an SA-β-GAL assay to detect senescent cell accumulation across the entire adult zebrafish organism ( Danio rerio ). We also identify previously unreported organs that show increased cell senescence in telomerase mutants, including the liver and the spinal cord.
Genes / Markers
Figures
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping