PUBLICATION

The identification of zebrafish mutants showing alterations in senescence-associated biomarkers

Authors
Kishi, S., Bayliss, P.E., Uchiyama, J., Koshimizu, E., Qi, J., Nanjappa, P., Imamura, S., Islam, A., Neuberg, D., Amsterdam, A., and Roberts, T.M.
ID
ZDB-PUB-080826-21
Date
2008
Source
PLoS Genetics   4(8): e1000152 (Journal)
Registered Authors
Amsterdam, Adam, Bayliss, Peter, Kishi, Shuji, Roberts, Thomas M., Uchiyama, Junzo
Keywords
Embryos, Zebrafish, Phenotypes, Oxidative stress, Retina, Genetic screens, Biomarkers, Eyes
MeSH Terms
  • Aging/genetics
  • Aging/metabolism*
  • Animals
  • Biomarkers/analysis
  • Gene Expression
  • Humans
  • Longevity
  • Mutagenesis, Insertional*
  • Oxidative Stress
  • Telomeric Repeat Binding Protein 2/genetics
  • Telomeric Repeat Binding Protein 2/metabolism
  • Zebrafish/embryology
  • Zebrafish/genetics*
  • Zebrafish/metabolism*
  • Zebrafish Proteins/genetics
  • Zebrafish Proteins/metabolism*
  • beta-Galactosidase/genetics
  • beta-Galactosidase/metabolism*
(all 18)
PubMed
18704191 Full text @ PLoS Genet.
Abstract
There is an interesting overlap of function in a wide range of organisms between genes that modulate the stress responses and those that regulate aging phenotypes and, in some cases, lifespan. We have therefore screened mutagenized zebrafish embryos for the altered expression of a stress biomarker, senescence-associated beta-galactosidase (SA-beta-gal) in our current study. We validated the use of embryonic SA-beta-gal production as a screening tool by analyzing a collection of retrovirus-insertional mutants. From a pool of 306 such mutants, we identified 11 candidates that showed higher embryonic SA-beta-gal activity, two of which were selected for further study. One of these mutants is null for a homologue of Drosophila spinster, a gene known to regulate lifespan in flies, whereas the other harbors a mutation in a homologue of the human telomeric repeat binding factor 2 (terf2) gene, which plays roles in telomere protection and telomere-length regulation. Although the homozygous spinster and terf2 mutants are embryonic lethal, heterozygous adult fish are viable and show an accelerated appearance of aging symptoms including lipofuscin accumulation, which is another biomarker, and shorter lifespan. We next used the same SA-beta-gal assay to screen chemically mutagenized zebrafish, each of which was heterozygous for lesions in multiple genes, under the sensitizing conditions of oxidative stress. We obtained eight additional mutants from this screen that, when bred to homozygosity, showed enhanced SA-beta-gal activity even in the absence of stress, and further displayed embryonic neural and muscular degenerative phenotypes. Adult fish that are heterozygous for these mutations also showed the premature expression of aging biomarkers and the accelerated onset of aging phenotypes. Our current strategy of mutant screening for a senescence-associated biomarker in zebrafish embryos may thus prove to be a useful new tool for the genetic dissection of vertebrate stress response and senescence mechanisms.
Genes / Markers
Marker Marker Type Name
atp6v0caGENEATPase H+ transporting V0 subunit ca
atp6v1hGENEATPase H+ transporting V1 subunit H
catGENEcatalase
clint1aGENEclathrin interactor 1a
cnot1GENECCR4-NOT transcription complex, subunit 1
dmdGENEdystrophin
dtlGENEdenticleless E3 ubiquitin protein ligase homolog (Drosophila)
eif3dGENEeukaryotic translation initiation factor 3, subunit D
polr2dGENERNA polymerase II subunit D
polr2gGENERNA polymerase II subunit G
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Figures
Figure Gallery (16 images) / 2
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Expression
Phenotype
Mutations / Transgenics
Allele Construct Type Affected Genomic Region
hi229TgTransgenic Insertion
hi447TgTransgenic Insertion
hi601TgTransgenic Insertion
hi891TgTransgenic Insertion
hi923TgTransgenic Insertion
hi1113aTgTransgenic Insertion
hi1182TgTransgenic Insertion
hi1207TgTransgenic Insertion
hi1520TgTransgenic Insertion
hi1640TgTransgenic Insertion
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Human Disease / Model
No data available
Sequence Targeting Reagents
Target Reagent Reagent Type
catMO1-catMRPHLNO
spns1MO1-spns1MRPHLNO
terfaMO1-terfaMRPHLNO
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Fish
Antibodies
Name Type Antigen Genes Isotypes Host Organism
Ab1-dmdmonoclonalIgG1Mouse
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Orthology
No data available
Engineered Foreign Genes
No data available
Mapping
No data available