Adhesion Receptor Adgrg2/Gpr64 Is In The Gi-Tract Selectively Expressed In Mature Intestinal Tuft Cells.

Abstract Objective GPR64/ADGRG2 is an orphan Adhesion G protein-coupled receptor (ADGR) known mainly to be expressed in parathyroid gland and epididymis. The aim of this investigation was to delineate the cellular expression of GPR64 throughout the body with focus on the GI tract. Methods Transgenic Gpr64mCherry reporter mice were histologically examined throughout the body and reporter protein expression in intestinal tuft cells was confirmed by specific cell ablation. The GPCR repertoire of intestinal Gpr64mCherry-positive tuft cells was analyzed by quantitative RT-PCR analysis and in situ hybridization. The Gpr64mCherry was crossed into the general tuft cell reporter Trpm5GFP to generate small intestinal organoids for time-lapse imaging. Intestinal tuft cells were isolated from small intestine, FACS-purified and transcriptionally compared using RNA-seq analysis. Results Expression of the Gpr64mCherry reporter was identified in multiple organs and specifically in olfactory microvillous cells, enteric nerves, and importantly in respiratory and gastrointestinal tuft cells. In the small intestine, cell ablation targeting Gpr64-expressing epithelial cells eliminated tuft cells. Transcriptional analysis of small intestinal Gpr64mCherry -positive tuft cells confirmed expression of Gpr64 and the chemo-sensors Sucnr1, Gprc5c, Drd3, and Gpr41/Ffar3. Time-lapse studies of organoids from Trpm5GFP:Gpr64mCherry mice revealed sequential expression of initially Trpm5GFP and subsequently also Gpr64mCherry in maturing intestinal tuft cells. RNA-seq analysis of small intestinal tuft cells based on these two markers demonstrated a dynamic change in expression of transcription factors and GPCRs from young to mature tuft cells. Conclusions GPR64 is expressed in chemosensory epithelial cells across a broad range of tissues but in the GI tract remarkedly selectively in mature versus young immunoregulatory tuft cells.