Compounds excreted by Caenorhabditis elegans at 4°C alleviate gamma-radiation injury through the gcy-5-mediated lysosomal pathway
Description
In this work, C. elegans was found to withstand high-dose radiation when exposed to 4°C for 4 hours before irradiation. After experimental confirmation, it was found that compounds excreted by C. elegans at 4°C inhibited radiation injury. The excreted compounds targeted the lysosomal pathway mediated by guanylyl cyclase gcy-5. Furthermore, activation of gcy-5 by the excreted compounds inhibited the expression of fat-5, which suppresses the lysosomal pathway.
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To investigate lysosomal function by fluorescence staining, LysoTracker Red (#C1046, Beyotime, Shanghai, China) and LysoSensor Green (LysoSensor™ Green DND-189, #L7535, Invitrogen, USA) were used. Worms in the L1 stage were treated as indicated. Twenty-four hours after irradiation, the worms were collected and washed with M9 buffer. LysoTracker Red and LysoSensor Green were diluted as indicated by the manufacturer’s instructions. The worms were stained with these LysoTracker Red and LysoSensor Green staining solutions for 30 mins at 37°C following the manufacturer’s instructions. The LysoSensor Green signal was observed at wavelengths of 500 nm~545 nm with 488 nm excitation. The LysoTracker Red signals was observed at wavelengths of 570 nm~610 nm with 552 nm excitation.
Institutions
- Chinese Academy of Medical Sciences Institute of Radiation Medicine