Long-term inhibition of cyclophilin D results in intracellular translocation of calcein AM from mitochondria to lysosomes

Cyclophilin D is a peptidyl-prolyl cis-trans isomerase localized in the mitochondrial matrix. Although its effects on mitochondrial characteristics have been well studied, its relation to the uptake of molecules by mitochondria remains unknown. Here, we demonstrated the effects of cyclophilin D on the intracellular translocation of calcein AM. Following addition of calcein AM to control cells or cells overexpressing wild-type cyclophilin D, calcein fluorescence was observed in mitochondria. However, long-term inhibition of cyclophilin D in these cells altered the localization of calcein fluorescence from mitochondria to lysosomes without changing mitochondrial esterase activity. In addition, depletion of glucose from the medium recovered calcein localization from lysosomes to mitochondria. This is the first demonstration of the effects of cyclophilin D on the intracellular translocation of molecules other than proteins and suggests that cyclophilin D may modify mitochondrial features by inducing the translocation of molecules to the mitochondria through the mechanism associated with cellular energy metabolism. [Display omitted] •Calcein AM enters the mitochondria in C6 and HeLa cells.•Calcein AM enters lysosomes upon long-term inhibition of cyclophilin D.•Transport of calcein AM to lysosomes depends on cellular energy metabolism.•Translocation to lysosomes occurs in a cargo-specific manner.