論文

査読有り 国際誌
2020年1月29日

Visualization of a blue light transmission area in living animals using light-induced nuclear translocation of fluorescent proteins.

Biochemical and biophysical research communications
  • Ayumu Inutsuka
  • ,
  • Norihiro Kimizuka
  • ,
  • Natsuki Takanohashi
  • ,
  • Hisashi Yakabu
  • ,
  • Tatsushi Onaka

522
1
開始ページ
138
終了ページ
143
記述言語
英語
掲載種別
研究論文(学術雑誌)
DOI
10.1016/j.bbrc.2019.11.023

Optical manipulations are widely used to analyze neuronal functions in vivo. Blue light is frequently used to activate channelrhodopsins or LOV domains, although the degrees of its absorption and scattering are higher than those of longer wavelength light. High spatial resolution of optical manipulation is easily achieved in vitro, while the light is unevenly scattered and absorbed in tissues due to many factors. It is difficult to spatially measure a blue light transmission area in vivo. Here, we propose a genetic method to visualize blue light transmission in the brain and other organs using light-induced nuclear translocation of fluorescent proteins with a LOV domain. A light-inducible nuclear localization signal (LINuS) consists of a LOV2 domain fused with a nuclear localization signal (NLS). We confirmed that blue light illumination induced reversible translocation of NES-tdTomato-LINuS from the cytosol to the nucleus within 30 min in HEK293 cells. By employing a PHP.eb capsid that can penetrate the blood-brain barrier, retro-orbital sinus injection of adeno-associated virus (AAV) vectors induced scattered expression of nuclear export signal (NES)-tdTomato-LINuS in the brain. We confirmed that 30-min transcranial blue light illumination induced nuclear translocation of NES-tdTomato-LINuS in the cortex, the hippocampus, and even the paraventricular nucleus of the thalamus. We also found that mice exposed to blue light in a shaved abdominal area exhibited a substantial increase in nuclear translocation in the ventral surface lobe of the liver. These results provide a simple way to obtain useful information on light transmission in tissues without any transgenic animals or skillful procedures.

リンク情報
DOI
https://doi.org/10.1016/j.bbrc.2019.11.023
PubMed
https://www.ncbi.nlm.nih.gov/pubmed/31757418
ID情報
  • DOI : 10.1016/j.bbrc.2019.11.023
  • PubMed ID : 31757418

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