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Analysis of cataract-regulated genes using chemical DNA damage induction in a rat ex vivo model

Although cataracts affect almost all people at advanced age and carry a risk of blindness, the mechanisms of cataract development remain incompletely understood. Oxidative stress, which is a causative factor in cataract, results in DNA breakage, which suggests that DNA damage could contribute to the...

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Autores principales: Yamaoka, Risa, Kanada, Fumito, Nagaya, Masaya, Takashima, Masaru, Takamura, Yoshihiro, Inatani, Masaru, Oki, Masaya
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9728860/
https://www.ncbi.nlm.nih.gov/pubmed/36477544
http://dx.doi.org/10.1371/journal.pone.0273456
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author Yamaoka, Risa
Kanada, Fumito
Nagaya, Masaya
Takashima, Masaru
Takamura, Yoshihiro
Inatani, Masaru
Oki, Masaya
author_facet Yamaoka, Risa
Kanada, Fumito
Nagaya, Masaya
Takashima, Masaru
Takamura, Yoshihiro
Inatani, Masaru
Oki, Masaya
author_sort Yamaoka, Risa
collection PubMed
description Although cataracts affect almost all people at advanced age and carry a risk of blindness, the mechanisms of cataract development remain incompletely understood. Oxidative stress, which is a causative factor in cataract, results in DNA breakage, which suggests that DNA damage could contribute to the formation of cataracts. We developed an ex vivo experimental system to study changes in gene expression during the formation of opacities in the lens by culturing explanted rat lenses with Methylmethanesulfonate (MMS) or Bleomycin, which induce DNA damage. Lenses cultured using this experimental system developed cortical opacity, which increased in a concentration- and time-dependent manner. In addition, we compared expression profiles at the whole gene level using microarray analysis of lenses subjected to MMS or Bleomycin stress. Microarray findings in MMS-induced opacity were validated and gene expression was measured from Days 1–4 using RT-qPCR. Altered genes were classified into four groups based on the days of peak gene expression: Group 1, in which expression peaked on Day 1; Group 2, in which expression peaked on Day 2; Group 3, in which expression progressively increased from Days 1–4 or were upregulated on Day 1 and sustained through Day 4; and Group 4, in which expression level oscillated from Days 1–4. Genes involved in lipid metabolism were restricted to Group 1. DNA repair- and cell cycle-related genes were restricted to Groups 1 and 2. Genes associated with oxidative stress and drug efflux were restricted to Group 2. These findings suggest that in temporal changes of MMS-induced opacity formation, the activated pathways could occur in the following order: lipid metabolism, DNA repair and cell cycle, and oxidative stress and drug efflux.
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spelling pubmed-97288602022-12-08 Analysis of cataract-regulated genes using chemical DNA damage induction in a rat ex vivo model Yamaoka, Risa Kanada, Fumito Nagaya, Masaya Takashima, Masaru Takamura, Yoshihiro Inatani, Masaru Oki, Masaya PLoS One Research Article Although cataracts affect almost all people at advanced age and carry a risk of blindness, the mechanisms of cataract development remain incompletely understood. Oxidative stress, which is a causative factor in cataract, results in DNA breakage, which suggests that DNA damage could contribute to the formation of cataracts. We developed an ex vivo experimental system to study changes in gene expression during the formation of opacities in the lens by culturing explanted rat lenses with Methylmethanesulfonate (MMS) or Bleomycin, which induce DNA damage. Lenses cultured using this experimental system developed cortical opacity, which increased in a concentration- and time-dependent manner. In addition, we compared expression profiles at the whole gene level using microarray analysis of lenses subjected to MMS or Bleomycin stress. Microarray findings in MMS-induced opacity were validated and gene expression was measured from Days 1–4 using RT-qPCR. Altered genes were classified into four groups based on the days of peak gene expression: Group 1, in which expression peaked on Day 1; Group 2, in which expression peaked on Day 2; Group 3, in which expression progressively increased from Days 1–4 or were upregulated on Day 1 and sustained through Day 4; and Group 4, in which expression level oscillated from Days 1–4. Genes involved in lipid metabolism were restricted to Group 1. DNA repair- and cell cycle-related genes were restricted to Groups 1 and 2. Genes associated with oxidative stress and drug efflux were restricted to Group 2. These findings suggest that in temporal changes of MMS-induced opacity formation, the activated pathways could occur in the following order: lipid metabolism, DNA repair and cell cycle, and oxidative stress and drug efflux. Public Library of Science 2022-12-07 /pmc/articles/PMC9728860/ /pubmed/36477544 http://dx.doi.org/10.1371/journal.pone.0273456 Text en © 2022 Yamaoka et al https://creativecommons.org/licenses/by/4.0/This is an open access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
Yamaoka, Risa
Kanada, Fumito
Nagaya, Masaya
Takashima, Masaru
Takamura, Yoshihiro
Inatani, Masaru
Oki, Masaya
Analysis of cataract-regulated genes using chemical DNA damage induction in a rat ex vivo model
title Analysis of cataract-regulated genes using chemical DNA damage induction in a rat ex vivo model
title_full Analysis of cataract-regulated genes using chemical DNA damage induction in a rat ex vivo model
title_fullStr Analysis of cataract-regulated genes using chemical DNA damage induction in a rat ex vivo model
title_full_unstemmed Analysis of cataract-regulated genes using chemical DNA damage induction in a rat ex vivo model
title_short Analysis of cataract-regulated genes using chemical DNA damage induction in a rat ex vivo model
title_sort analysis of cataract-regulated genes using chemical dna damage induction in a rat ex vivo model
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9728860/
https://www.ncbi.nlm.nih.gov/pubmed/36477544
http://dx.doi.org/10.1371/journal.pone.0273456
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