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Tango knock-ins visualize endogenous activity of G protein-coupled receptors in Drosophila
Authors:Hidetaka Katow  Takahiro Takahashi  Kuniaki Saito  Hiromu Tanimoto
Affiliation:1. Genetic Strains Research Center, National Institute of Genetics, Mishima, Japan;2. Graduate School of Life Sciences, Tohoku University, Sendai, Japan
Abstract:
G protein-coupled receptors (GPCRs) represent a family of seven-pass transmembrane protein receptors whose ligands include neuropeptides and small-molecule neuromodulators such as dopamine and serotonin. These neurotransmitters act at long distances and are proposed to define the ground state of the nervous system. The Drosophila genome encodes approximately 50 neuropeptides and their functions in physiology and behavior are now under intensive studies. Key information currently lacking in the field is the spatiotemporal activation patterns of endogenous GPCRs. Here we report application of the Tango system, a reporter assay to detect GPCR activity, to endogenous GPCRs in the fly genome. We developed a method to integrate the sensor component of the Tango system to the C-terminus of endogenous genes by using genome editing techniques. We demonstrate that Tango sensors in the Sex-peptide receptor (SPR) locus allow sensitive detection of mating-dependent SPR activity in the female reproductive organ. The method is easily applicable to any GPCR and will provide a way to systematically characterize GPCRs in the fly brain.
Keywords:Drosophila  G protein-coupled receptor  tango assay  sex-peptide receptor
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