Optogenetic in vivo cell manipulation in KillerRed-expressing zebrafish transgenics

Teh, C., Chudakov, D.M., Poon, K.L., Mamedov, I.Z., Sek, J.Y., Shidlovsky, K., Lukyanov, S., and Korzh, V.
BMC Developmental Biology   10: 110 (Journal)
Registered Authors
Korzh, Vladimir, Poon, Kar Lai, Sek, Jun-Yan, Teh, Cathleen
MeSH Terms
  • Animals
  • Animals, Genetically Modified*/anatomy & histology
  • Animals, Genetically Modified*/embryology
  • Animals, Genetically Modified*/genetics
  • Apoptosis
  • DNA Transposable Elements
  • Embryo, Nonmammalian*/anatomy & histology
  • Embryo, Nonmammalian*/drug effects
  • Embryo, Nonmammalian*/physiology
  • Enhancer Elements, Genetic
  • Green Fluorescent Proteins/genetics
  • Green Fluorescent Proteins/pharmacology*
  • Humans
  • In Situ Nick-End Labeling
  • Microscopy, Fluorescence/methods*
  • Morphogenesis/drug effects
  • Photosensitizing Agents/pharmacology*
  • Reactive Oxygen Species/pharmacology
  • Zebrafish*/anatomy & histology
  • Zebrafish*/embryology
  • Zebrafish*/genetics
21040591 Full text @ BMC Dev. Biol.
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.
Genes / Markers
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Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Engineered Foreign Genes