摘要
A two-photon ratiometric fluorescent sensor for Cu^+ in endoplasmic reticulum(ER), CNSB, was developed via coumarin/ASBD integration based on FRET mechanism. In solution, CNSB shows reversible, highly-specific ratiometric response to Cu^+ .Moreover, CNSB exhibits suitable K_d value, suggesting the possibility of detecting Cu^+ in the living cells. The probe can enter the MCF-7 cells easily and specifically locates in the ER. The highly specific ratiometric response of CNSB toward Cu^+ in MCF-7 cells provides the sensor the capacity to visualize both exogenous and endogenous Cu^+ in the ER via fluorescence imaging.Next, CNSB was utilized to detect the fluctuation and distribution of Cu^+ under ER stress in MCF-7 cells, which confirmed directly the relationship between Cu^+ enhancement and ER stress. Meanwhile, the two-photon ability of coumarin facilitated the sensor to visualize Cu^+ fluctuation via two-photon fluorescence imaging. In addition, the spatial distribution of Cu^+ in the heart slice of the 14-day-old rat was demonstrated using CNSB. This study demonstrates the promising potential of CNSB in clarifying the Cu^+ -dependent signaling in the ER stress-related diseases.
A two-photon ratiometric fluorescent sensor for Cu^+ in endoplasmic reticulum(ER), CNSB, was developed via coumarin/ASBD integration based on FRET mechanism. In solution, CNSB shows reversible, highly-specific ratiometric response to Cu^+ .Moreover, CNSB exhibits suitable K_d value, suggesting the possibility of detecting Cu^+ in the living cells. The probe can enter the MCF-7 cells easily and specifically locates in the ER. The highly specific ratiometric response of CNSB toward Cu^+ in MCF-7 cells provides the sensor the capacity to visualize both exogenous and endogenous Cu^+ in the ER via fluorescence imaging.Next, CNSB was utilized to detect the fluctuation and distribution of Cu^+ under ER stress in MCF-7 cells, which confirmed directly the relationship between Cu^+ enhancement and ER stress. Meanwhile, the two-photon ability of coumarin facilitated the sensor to visualize Cu^+ fluctuation via two-photon fluorescence imaging. In addition, the spatial distribution of Cu^+ in the heart slice of the 14-day-old rat was demonstrated using CNSB. This study demonstrates the promising potential of CNSB in clarifying the Cu^+ -dependent signaling in the ER stress-related diseases.
基金
supported by the National Basic Research Program of China (2015CB856300)
the National Natural Science Foundation of China (21571099, 21731004)
the Natural Science Foundation of Jiangsu (BK20150054)
