Endoplasmic reticulum targeted fluorescent probe for real-time monitoring the viscosity changes induced by calcium homeostasis disruption.

ABSTRACT

The endoplasmic reticulum (ER) acts as the major storage site for calcium ions, which are messenger ions for intracellular signaling. Disruption of calcium ion homeostasis can significantly affect the viscosity, polarity and pH of the ER. However, it is still unclear the relationship between the viscosity changes in ER and the imbalance of calcium ion homeostasis. Herein, we developed a novel fluorescent probe, named TPA, for monitoring viscosity changes that specifically targets the endoplasmic reticulum rather than mitochondria or lysosomes. TPA probe displayed good stability, as well as high responsiveness and selectivity to viscosity. The fluorescence intensity of TPA was significantly enhanced with the increased concentration or incubation time of the stimulating agents(i.e., tunicamycin), showing high responsiveness to the viscosity changes in ER. Furthermore, the TPA probe successfully demonstrated that an increase in intracellular calcium ion concentration leads to an increase in ER viscosity, whereas a decrease in calcium ion concentration leads to a decrease viscosity in ER. Both in vitro and in vivo experiments demonstrated that TPA probe successfully detected the viscosity changes in ER, especially the effects of calcium ion homeostasis disruption on ER. Overall, the TPA probe represents an efficient method for studying the relationship between calcium ion homeostasis and ER viscosity.