NanoBRET in C. elegans illuminates functional receptor interactions in real time

Abstract

Background Protein-protein interactions form the basis of every organism and thus, investigating their dynamics, intracellular protein localization, trafficking and interactions of distinct proteins such as receptors and their ligand-binding are of general interest. Bioluminescence resonance energy transfer (BRET) is a powerful tool to investigate these aspects in vitro. Since in vitro approaches mostly neglect the more complex in vivo situation, we established BRET as an in vivo tool for studying protein interactions in the nematode C. elegans. Results We generated worms expressing NanoBRET sensors and elucidated the interaction of two ligand-G protein-coupled receptor (GPCR) pairs, the neuropeptide receptor NPR-11 and the Adhesion GPCR LAT-1. Furthermore, we adapted the enhanced bystander BRET technology to measure subcellular protein localization. Using this approach, we traced ligand-induced internalization of NPR-11 in vivo. Conclusions Our results indicate that in vivo NanoBRET is a tool to investigate specific protein interactions and localization in a physiological setting in real time in the living organism C. elegans.

Bioluminescence as a functional tool for visualizing and controlling neuronal activity in vivo

“… 120 Likewise, the exoskeleton and the cuticle in invertebrates is often encountered as a diffusion barrier that needs to be overcome. 121 , 122 Thus, the bioluminescent signal may not report the cellular activity but the distribution of the cofactor, requiring luciferin delivery optimization for different tissues. In C. elegans , we consistently observed bioluminescence in amphid cells (ASH and AWA) or the vulva regions that are exposed to the environment under conditions in which we did not observe signal in other tissues.…”