Generation and characterization of keap1a- and keap1b-knockout zebrafish
The Keap1–Nrf2 pathway is an evolutionarily conserved mechanism that protects cells from oxidative stress and electrophiles. Under homeostatic conditions, Keap1 interacts with Nrf2 and leads to its rapid proteasomal degradation, but when cells are exposed to oxidative stress/electrophiles, Keap1 sen...
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doaj-5dd5559339e84a2aa4e6901160c6bb392020-11-25T03:19:16ZengElsevierRedox Biology2213-23172020-09-0136101667Generation and characterization of keap1a- and keap1b-knockout zebrafishVu Thanh Nguyen0Lixuan Bian1Junya Tamaoki2Shiro Otsubo3Masafumi Muratani4Atsuo Kawahara5Makoto Kobayashi6Department of Molecular and Developmental Biology, Faculty of Medicine, University of Tsukuba, 1-1-1 Tennodai, Tsukuba, 305-8575, Japan; Division of Aquaculture Biotechnology, Biotechnology Center of Ho Chi Minh City, Ho Chi Minh City, Viet NamDepartment of Molecular and Developmental Biology, Faculty of Medicine, University of Tsukuba, 1-1-1 Tennodai, Tsukuba, 305-8575, JapanDepartment of Molecular and Developmental Biology, Faculty of Medicine, University of Tsukuba, 1-1-1 Tennodai, Tsukuba, 305-8575, JapanDepartment of Molecular and Developmental Biology, Faculty of Medicine, University of Tsukuba, 1-1-1 Tennodai, Tsukuba, 305-8575, JapanDepartment of Genome Biology, Faculty of Medicine, University of Tsukuba, 1-1-1 Tennodai, Tsukuba, 305-8575, JapanLaboratory for Developmental Biology, Center for Medical Education and Sciences, Graduate School of Medical Science, University of Yamanashi, Chuo, Yamanashi, 409-3898, JapanDepartment of Molecular and Developmental Biology, Faculty of Medicine, University of Tsukuba, 1-1-1 Tennodai, Tsukuba, 305-8575, Japan; Corresponding author.The Keap1–Nrf2 pathway is an evolutionarily conserved mechanism that protects cells from oxidative stress and electrophiles. Under homeostatic conditions, Keap1 interacts with Nrf2 and leads to its rapid proteasomal degradation, but when cells are exposed to oxidative stress/electrophiles, Keap1 senses them, resulting in an improper Keap1–Nrf2 interaction and Nrf2 stabilization. Keap1 is therefore considered both an “inhibitor” of and “stress sensor” for Nrf2 activation. Interestingly, fish and amphibians have two Keap1s (Keap1a and Keap1b), while there is only one in mammals, birds and reptiles. A phylogenetic analysis suggested that mammalian Keap1 is an ortholog of fish Keap1b, not Keap1a. In this study, we investigated the differences and similarities between Keap1a and Keap1b using zebrafish genetics. We generated zebrafish knockout lines of keap1a and keap1b. Homozygous mutants of both knockout lines were viable and fertile. In both mutant larvae, the basal expression of Nrf2 target genes and antioxidant activity were up-regulated in an Nrf2-dependent manner, suggesting that both Keap1a and Keap1b can function as Nrf2 inhibitors. We also analyzed the effects of the Nrf2 activator sulforaphane in these mutants and found that keap1a-, but not keap1b-, knockout larvae responded to sulforaphane, suggesting that the stress/chemical-sensing abilities of the two Keap1s are different.http://www.sciencedirect.com/science/article/pii/S2213231720308727Antioxidant activityKeap1–Nrf2 pathwayKnockout zebrafishOxidative stressSulforaphane |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Vu Thanh Nguyen Lixuan Bian Junya Tamaoki Shiro Otsubo Masafumi Muratani Atsuo Kawahara Makoto Kobayashi |
spellingShingle |
Vu Thanh Nguyen Lixuan Bian Junya Tamaoki Shiro Otsubo Masafumi Muratani Atsuo Kawahara Makoto Kobayashi Generation and characterization of keap1a- and keap1b-knockout zebrafish Redox Biology Antioxidant activity Keap1–Nrf2 pathway Knockout zebrafish Oxidative stress Sulforaphane |
author_facet |
Vu Thanh Nguyen Lixuan Bian Junya Tamaoki Shiro Otsubo Masafumi Muratani Atsuo Kawahara Makoto Kobayashi |
author_sort |
Vu Thanh Nguyen |
title |
Generation and characterization of keap1a- and keap1b-knockout zebrafish |
title_short |
Generation and characterization of keap1a- and keap1b-knockout zebrafish |
title_full |
Generation and characterization of keap1a- and keap1b-knockout zebrafish |
title_fullStr |
Generation and characterization of keap1a- and keap1b-knockout zebrafish |
title_full_unstemmed |
Generation and characterization of keap1a- and keap1b-knockout zebrafish |
title_sort |
generation and characterization of keap1a- and keap1b-knockout zebrafish |
publisher |
Elsevier |
series |
Redox Biology |
issn |
2213-2317 |
publishDate |
2020-09-01 |
description |
The Keap1–Nrf2 pathway is an evolutionarily conserved mechanism that protects cells from oxidative stress and electrophiles. Under homeostatic conditions, Keap1 interacts with Nrf2 and leads to its rapid proteasomal degradation, but when cells are exposed to oxidative stress/electrophiles, Keap1 senses them, resulting in an improper Keap1–Nrf2 interaction and Nrf2 stabilization. Keap1 is therefore considered both an “inhibitor” of and “stress sensor” for Nrf2 activation. Interestingly, fish and amphibians have two Keap1s (Keap1a and Keap1b), while there is only one in mammals, birds and reptiles. A phylogenetic analysis suggested that mammalian Keap1 is an ortholog of fish Keap1b, not Keap1a. In this study, we investigated the differences and similarities between Keap1a and Keap1b using zebrafish genetics. We generated zebrafish knockout lines of keap1a and keap1b. Homozygous mutants of both knockout lines were viable and fertile. In both mutant larvae, the basal expression of Nrf2 target genes and antioxidant activity were up-regulated in an Nrf2-dependent manner, suggesting that both Keap1a and Keap1b can function as Nrf2 inhibitors. We also analyzed the effects of the Nrf2 activator sulforaphane in these mutants and found that keap1a-, but not keap1b-, knockout larvae responded to sulforaphane, suggesting that the stress/chemical-sensing abilities of the two Keap1s are different. |
topic |
Antioxidant activity Keap1–Nrf2 pathway Knockout zebrafish Oxidative stress Sulforaphane |
url |
http://www.sciencedirect.com/science/article/pii/S2213231720308727 |
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