RRC ID 60760
Author Katayama H, Hama H, Nagasawa K, Kurokawa H, Sugiyama M, Ando R, Funata M, Yoshida N, Homma M, Nishimura T, Takahashi M, Ishida Y, Hioki H, Tsujihata Y, Miyawaki A.
Title Visualizing and Modulating Mitophagy for Therapeutic Studies of Neurodegeneration.
Journal Cell
Abstract Dysfunctional mitochondria accumulate in many human diseases. Accordingly, mitophagy, which removes these mitochondria through lysosomal degradation, is attracting broad attention. Due to uncertainties in the operational principles of conventional mitophagy probes, however, the specificity and quantitativeness of their readouts are disputable. Thorough investigation of the behaviors and fates of fluorescent proteins inside and outside lysosomes enabled us to develop an indicator for mitophagy, mito-SRAI. Through strict control of its mitochondrial targeting, we were able to monitor mitophagy in fixed biological samples more reproducibly than before. Large-scale image-based high-throughput screening led to the discovery of a hit compound that induces selective mitophagy of damaged mitochondria. In a mouse model of Parkinsons disease, we found that dopaminergic neurons selectively failed to execute mitophagy that promoted their survival within lesions. These results show that mito-SRAI is an essential tool for quantitative studies of mitochondrial quality control.
Volume 181(5)
Pages 1176-1187.e16
Published 2020-5-28
DOI 10.1016/j.cell.2020.04.025
PII S0092-8674(20)30488-8
PMID 32437660
MeSH Animals Autophagy / physiology Fluorescence Resonance Energy Transfer / methods* Fluorescent Antibody Technique / methods Fluorescent Dyes / chemistry Humans Lysosomes / metabolism* Lysosomes / physiology Male Mice Mice, Inbred C57BL Mitochondria / metabolism Mitophagy / genetics Mitophagy / physiology*
IF 38.637
Times Cited 1
DNA material SRAI_pcDNA3 (RDB18222) mito-SRAI_pcDNA3 (RDB18223) pAAV2-TRE-mito-SRAI-BGHpA (RDB18684) pAAV2-SYN-tTAad-BGHpA (RDB18685) pcDNA3/mCherry-Parkin (RDB20336)