RRC ID 30804
Author Kimura E, Deguchi T, Kamei Y, Shoji W, Yuba S, Hitomi J.
Title Application of infrared laser to the zebrafish vascular system: gene induction, tracing, and ablation of single endothelial cells.
Journal Arterioscler Thromb Vasc Biol
Abstract OBJECTIVE:Infrared laser-evoked gene operator is a new microscopic method optimized to heat cells in living organisms without causing photochemical damage. By combining the promoter system for the heat shock response, infrared laser-evoked gene operator enables laser-mediated gene induction in targeted cells. We applied this method to the vascular system in zebrafish embryos and demonstrated its usability to investigate mechanisms of vascular morphogenesis in vivo.
APPROACH AND RESULTS:We used double-transgenic zebrafish with fli1:nEGFP to identify the endothelial cells, and with hsp:mCherry to carry out single-cell labeling. Optimizing the irradiation conditions, we finally succeeded in inducing the expression of the mCherry gene in single targeted endothelial cells, at a maximum efficiency rate of 60%. In addition, we indicated that this system could be used for laser ablation under certain conditions. To evaluate infrared laser-evoked gene operator, we applied this system to the endothelial cells of the first intersegmental arteries, and captured images of the connection between the vascular systems of the brain and spinal cord.
CONCLUSIONS:Our results suggest that the infrared laser-evoked gene operator system will contribute to the elucidation of the mechanisms underlying vascular morphogenesis by controlling spatiotemporal gene activation in single endothelial cells, by labeling or deleting individual vessels in living embryos.
Volume 33(6)
Pages 1264-70
Published 2013-6-1
DOI 10.1161/ATVBAHA.112.300602
PII ATVBAHA.112.300602
PMID 23539214
MeSH Animals Animals, Genetically Modified Blood Vessels / embryology* Endothelial Cells / radiation effects* Gene Expression Heat-Shock Response* Infrared Rays* Lasers Models, Animal Neovascularization, Physiologic / genetics Sensitivity and Specificity Transcriptional Activation* Zebrafish
IF 6.604
Times Cited 11
Paramecium KNZ1207 (PC122021A) Yad1g1N (PB031010B)