Press Release

Development of small fluorescent probes for the analysis of autophagy kinetics

July 6, 2023


Development of a red fluorescent probe for monitoring autophagy

Combinational use of the probes allows visualization of autophagy kinetics

The probes also detect the alternative autophagic compartments

Zebrafish stained with DAPRed represent autophagic activity in vivo

Autophagy is a dynamic process that degrades subcellular constituents, and its activity is measured by autophagic flux. The tandem proteins RFP-GFP-LC3 and GFP-LC3-RFP-LC3ΔG, which enable the visualization of autophagic vacuoles of different stages by differences in their fluorescent color, are useful tools to monitor autophagic flux, but they require plasmid transfection. In this study, we hence aimed to develop a new method to monitor autophagic flux using small cell-permeable fluorescent probes. We previously developed two green-fluorescent probes, DALGreen and DAPGreen, which detect autolysosomes and multistep autophagic vacuoles, respectively. We here developed a red-fluorescent autophagic probe, named DAPRed, which recognizes various autophagic vacuoles. By the combinatorial use of these green- and red-fluorescent probes, we were able to readily detect autophagic flux. Furthermore, these probes were useful not only for the visualization of canonical autophagy but also for alternative autophagy. DAPRed was also applicable for the detection of autophagy in living organisms.

Journal Article


TITLE:Development of small fluorescent probes for the analysis of autophagy kinetics


Correspondence to

Shigeomi Shimizu, M.D., Ph.D., Professor

Department of Pathological Cell Biology,
Medical Research Institute,
Tokyo Medical and Dental University (TMDU)

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