Live-cell microscopy of mitochondria-lysosome contacts and calcium dynamics

High spatial and temporal resolution live-cell microscopy was used to investigate the role of mitochondria-lysosome contacts in regulating mitochondrial calcium dynamics. The study examined how lysosomal calcium release through TRPML1 promotes calcium transfer to mitochondria at contact sites. The experiment involved real-time imaging of calcium dynamics using fluorescent indicators to visualize calcium transfer between lysosomes and mitochondria. The researchers used various pharmacological modulators and genetic manipulations to assess the contribution of specific channels including TRPML1, VDAC1, and the mitochondrial calcium uniporter to this process. The study demonstrated that mitochondrial calcium uptake at contact sites was dependent on tethering between the organelles and was modulated by both outer and inner mitochondrial membrane channels.