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Optogenetic in vivo cell manipulation in KillerRed-expressing zebrafish transgenics

BACKGROUND: KillerRed (KR) is a novel photosensitizer that efficiently generates reactive oxygen species (ROS) in KR-expressing cells upon intense green or white light illumination in vitro, resulting in damage to their plasma membrane and cell death. RESULTS: We report an in vivo modification of th...

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Autores principales: Teh, Cathleen, Chudakov, Dmitry M, Poon, Kar-Lai, Mamedov, Ilgar Z, Sek, Jun-Yan, Shidlovsky, Konstantin, Lukyanov, Sergey, Korzh, Vladimir
Formato: Texto
Lenguaje:English
Publicado: BioMed Central 2010
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2989954/
https://www.ncbi.nlm.nih.gov/pubmed/21040591
http://dx.doi.org/10.1186/1471-213X-10-110
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author Teh, Cathleen
Chudakov, Dmitry M
Poon, Kar-Lai
Mamedov, Ilgar Z
Sek, Jun-Yan
Shidlovsky, Konstantin
Lukyanov, Sergey
Korzh, Vladimir
author_facet Teh, Cathleen
Chudakov, Dmitry M
Poon, Kar-Lai
Mamedov, Ilgar Z
Sek, Jun-Yan
Shidlovsky, Konstantin
Lukyanov, Sergey
Korzh, Vladimir
author_sort Teh, Cathleen
collection PubMed
description BACKGROUND: KillerRed (KR) is a novel photosensitizer that efficiently generates reactive oxygen species (ROS) in KR-expressing cells upon intense green or white light illumination in vitro, resulting in damage to their plasma membrane and cell death. RESULTS: We report an in vivo modification of this technique using a fluorescent microscope and membrane-tagged KR (mem-KR)-expressing transgenic zebrafish. We generated several stable zebrafish Tol2 transposon-mediated enhancer-trap (ET) transgenic lines expressing mem-KR (SqKR series), and mapped the transposon insertion sites. As mem-KR accumulates on the cell membrane and/or Golgi, it highlights cell bodies and extensions, and reveals details of cellular morphology. The photodynamic property of KR made it possible to damage cells expressing this protein in a dose-dependent manner. As a proof-of-principle, two zebrafish transgenic lines were used to affect cell viability and function: SqKR2 expresses mem-KR in the hindbrain rhombomeres 3 and 5, and elsewhere; SqKR15 expresses mem-KR in the heart and elsewhere. Photobleaching of KR by intense light in the heart of SqKR15 embryos at lower levels caused a reduction in pumping efficiency of the heart and pericardial edema and at higher levels - in cell death in the hindbrain of SqKR2 and in the heart of SqKR15 embryos. CONCLUSIONS: An intense illumination of tissues expressing mem-KR affects cell viability and function in living zebrafish embryos. Hence, the zebrafish transgenics expressing mem-KR in a tissue-specific manner are useful tools for studying the biological effects of ROS.
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spelling pubmed-29899542010-12-13 Optogenetic in vivo cell manipulation in KillerRed-expressing zebrafish transgenics Teh, Cathleen Chudakov, Dmitry M Poon, Kar-Lai Mamedov, Ilgar Z Sek, Jun-Yan Shidlovsky, Konstantin Lukyanov, Sergey Korzh, Vladimir BMC Dev Biol Methodology Article BACKGROUND: KillerRed (KR) is a novel photosensitizer that efficiently generates reactive oxygen species (ROS) in KR-expressing cells upon intense green or white light illumination in vitro, resulting in damage to their plasma membrane and cell death. RESULTS: We report an in vivo modification of this technique using a fluorescent microscope and membrane-tagged KR (mem-KR)-expressing transgenic zebrafish. We generated several stable zebrafish Tol2 transposon-mediated enhancer-trap (ET) transgenic lines expressing mem-KR (SqKR series), and mapped the transposon insertion sites. As mem-KR accumulates on the cell membrane and/or Golgi, it highlights cell bodies and extensions, and reveals details of cellular morphology. The photodynamic property of KR made it possible to damage cells expressing this protein in a dose-dependent manner. As a proof-of-principle, two zebrafish transgenic lines were used to affect cell viability and function: SqKR2 expresses mem-KR in the hindbrain rhombomeres 3 and 5, and elsewhere; SqKR15 expresses mem-KR in the heart and elsewhere. Photobleaching of KR by intense light in the heart of SqKR15 embryos at lower levels caused a reduction in pumping efficiency of the heart and pericardial edema and at higher levels - in cell death in the hindbrain of SqKR2 and in the heart of SqKR15 embryos. CONCLUSIONS: An intense illumination of tissues expressing mem-KR affects cell viability and function in living zebrafish embryos. Hence, the zebrafish transgenics expressing mem-KR in a tissue-specific manner are useful tools for studying the biological effects of ROS. BioMed Central 2010-11-02 /pmc/articles/PMC2989954/ /pubmed/21040591 http://dx.doi.org/10.1186/1471-213X-10-110 Text en Copyright ©2010 Teh et al; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Methodology Article
Teh, Cathleen
Chudakov, Dmitry M
Poon, Kar-Lai
Mamedov, Ilgar Z
Sek, Jun-Yan
Shidlovsky, Konstantin
Lukyanov, Sergey
Korzh, Vladimir
Optogenetic in vivo cell manipulation in KillerRed-expressing zebrafish transgenics
title Optogenetic in vivo cell manipulation in KillerRed-expressing zebrafish transgenics
title_full Optogenetic in vivo cell manipulation in KillerRed-expressing zebrafish transgenics
title_fullStr Optogenetic in vivo cell manipulation in KillerRed-expressing zebrafish transgenics
title_full_unstemmed Optogenetic in vivo cell manipulation in KillerRed-expressing zebrafish transgenics
title_short Optogenetic in vivo cell manipulation in KillerRed-expressing zebrafish transgenics
title_sort optogenetic in vivo cell manipulation in killerred-expressing zebrafish transgenics
topic Methodology Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2989954/
https://www.ncbi.nlm.nih.gov/pubmed/21040591
http://dx.doi.org/10.1186/1471-213X-10-110
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