Imaging the nanomolar range of nitric oxide with an amplifier-coupled fluorescent indicator in living cells.
Nitric oxide (NO) is a small uncharged free radical that is involved in diverse physiological and pathophysiological mechanisms. NO is generated by three isoforms of NO synthase, endothelial, neuronal, and inducible ones. When generated in vascular endothelial cells, NO plays a key role in vascular tone regulation, in particular. Here, we describe an amplifier-coupled fluorescent indicator for NO to visualize physiological nanomolar dynamics of NO in living cells (detection limit of 0.1 nM). This genetically encoded high-sensitive indicator revealed that approximately 1 nM of NO, which is enough to relax blood vessels, is generated in vascular endothelial cells even in the absence of shear stress. The nanomolar range of basal endothelial NO thus revealed appears to be fundamental to vascular homeostasis.
Sato
Moritoshi
M
Department of Chemistry, School of Science, University of Tokyo, Tokyo 113-0033, Japan.
Hida
Naoki
N
Umezawa
Yoshio
Y
D000818:Animals; D016466:CHO Cells; D006224:Cricetinae; D003412:Cricetulus; D017931:DNA Primers; D042783:Endothelial Cells / Q000378:metabolism*; D007202:Indicators and Reagents / Q000378:metabolism; D008856:Microscopy, Fluorescence / Q000379:methods*; D008958:Models, Molecular; D009569:Nitric Oxide / Q000737:chemistry*; D010957:Plasmids / Q000235:genetics; D014162:Transfection
D017931:DNA Primers; D007202:Indicators and Reagents; D009569:Nitric Oxide