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  1. B300 農学部/生命農学研究科
  2. B300a 雑誌掲載論文
  3. 学術雑誌

Generation and evaluation of a transgenic zebrafish for tissue-specific expression of a dominant-negative Rho-associated protein kinase-2

http://hdl.handle.net/2237/00032620
http://hdl.handle.net/2237/00032620
49db3660-ad22-4c30-9a6b-c15a1fe85ad5
名前 / ファイル ライセンス アクション
Iida_et_al_2020_BBRC.pdf Iida_et_al_2020_BBRC (7.5 MB)
Item type 学術雑誌論文 / Journal Article(1)
公開日 2020-09-14
タイトル
タイトル Generation and evaluation of a transgenic zebrafish for tissue-specific expression of a dominant-negative Rho-associated protein kinase-2
言語 en
著者 Iida, Atsuo

× Iida, Atsuo

WEKO 101178

en Iida, Atsuo

Search repository
Wang, Zi

× Wang, Zi

WEKO 101179

en Wang, Zi

Search repository
Hondo, Eiichi

× Hondo, Eiichi

WEKO 101180

en Hondo, Eiichi

Search repository
Sehara-Fujisawa, Atsuko

× Sehara-Fujisawa, Atsuko

WEKO 101181

en Sehara-Fujisawa, Atsuko

Search repository
アクセス権
アクセス権 open access
アクセス権URI http://purl.org/coar/access_right/c_abf2
権利
言語 en
権利情報 © 2020. This manuscript version is made available under the CC-BY-NC-ND 4.0 license http://creativecommons.org/licenses/by-nc-nd/4.0/
キーワード
主題Scheme Other
主題 Rho pathway
キーワード
主題Scheme Other
主題 ROCK-2
キーワード
主題Scheme Other
主題 Dominant negative
キーワード
主題Scheme Other
主題 Zebrafish
キーワード
主題Scheme Other
主題 Cardiovascular development
抄録
内容記述 The Ras homologous (Rho) proteins are a family of small GTPases, which regulate the cytoskeleton and are related to stress fibers and focal adhesion. The Rho-associated protein kinases (ROCK) constitute part of the Rho effectors that regulate cell shape and movement via phosphorylation of the myosin light chain and actin depolymerizing factor/cofilin. ROCK members are widely expressed and play roles in various cell types during vertebrate development and morphogenesis; therefore, ROCK-knockout animals exhibit multiple defects mostly initiated at the embryonic stage. Analyzing the distinct roles of ROCK in cell shape and movement during the embryonic stages using live mammalian models is difficult. Here, we inhibited the Rho/ROCK pathway in zebrafish, which is a small fish that can be conveniently used as a developmental animal model in place of mammals. To inhibit the Rho/ROCK pathway, we designed a dominant-negative ROCK-2 (dnROCK-2) that lacked the kinase domain and was under the control of an upstream activation sequence (UAS). To evaluate the effects of expression of dnROCK-2, transgenic zebrafish lines were generated by mating strains expressing the construct with counterpart strains expressing the Gal4 activator in target tissues. In this study, we crossed the dnROCK-2-expressing line with two such Gal4-expressing lines; (1) SAGFF(LF)73A for expression in the whole body, and (2) Tg(fli1a: Gal4FF)^ubs4 for endothelial cell-specific expression. The phenotypes of the fish obtained were observed by fluorescent stereomicroscopy or confocal microscopy. Overexpression of dnROCK-2 in the whole body resulted in an inhibition of development, notably in cephalic formation, at 1-day post-fertilization (dpf). Confocal microscopy revealed that Hensen’s zone became unclear in the trunk muscle fibers expressing dnROCK-2. Endothelial cell-specific expression of dnROCK-2 caused abnormalities in cardiovascular formation at 2-dpf. These results suggest that dnROCK-2 can act as a dominant negative construct of the Rho/ROCK pathway to affect regulation of the cytoskeleton. This construct could be a convenient tool to investigate the function of ROCK members in other vertebrate cell types.
言語 en
内容記述タイプ Abstract
内容記述
内容記述 ファイル公開:2021-04-23
言語 ja
内容記述タイプ Other
出版者
言語 en
出版者 Elsevier
言語
言語 eng
資源タイプ
資源タイプresource http://purl.org/coar/resource_type/c_6501
タイプ journal article
出版タイプ
出版タイプ AM
出版タイプResource http://purl.org/coar/version/c_ab4af688f83e57aa
DOI
関連タイプ isVersionOf
識別子タイプ DOI
関連識別子 https://doi.org/10.1016/j.bbrc.2020.02.055
ISSN(print)
収録物識別子タイプ PISSN
収録物識別子 0006-291X
書誌情報 en : Biochemical and Biophysical Research Communications

巻 525, 号 1, p. 8-13, 発行日 2020-04-23
著者版フラグ
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