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Adhesion receptor ADGRG2/GPR64 is in the GI-tract selectively expressed in mature intestinal tuft cells
OBJECTIVE: GPR64/ADGRG2 is an orphan Adhesion G protein-coupled receptor (ADGR) known to be mainly expressed in the parathyroid gland and epididymis. This investigation aimed to delineate the cellular expression of GPR64 throughout the body with focus on the gastrointestinal (GI) tract. METHODS: Tra...
Autores principales: | , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Elsevier
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8105302/ https://www.ncbi.nlm.nih.gov/pubmed/33831593 http://dx.doi.org/10.1016/j.molmet.2021.101231 |
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author | Grunddal, Kaare V. Tonack, Sarah Egerod, Kristoffer L. Thompson, Jonathan James Petersen, Natalia Engelstoft, Maja S. Vagne, Constance Keime, Céline Gradwohl, Gérard Offermanns, Stefan Schwartz, Thue W. |
author_facet | Grunddal, Kaare V. Tonack, Sarah Egerod, Kristoffer L. Thompson, Jonathan James Petersen, Natalia Engelstoft, Maja S. Vagne, Constance Keime, Céline Gradwohl, Gérard Offermanns, Stefan Schwartz, Thue W. |
author_sort | Grunddal, Kaare V. |
collection | PubMed |
description | OBJECTIVE: GPR64/ADGRG2 is an orphan Adhesion G protein-coupled receptor (ADGR) known to be mainly expressed in the parathyroid gland and epididymis. This investigation aimed to delineate the cellular expression of GPR64 throughout the body with focus on the gastrointestinal (GI) tract. METHODS: Transgenic Gpr64(mCherry) 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 Gpr64(mCherry)-positive tuft cells was analyzed by quantitative RT-PCR analysis and in situ hybridization. The Gpr64(mCherry) was crossed into the general tuft cell reporter Trpm5(GFP) 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 Gpr64(mCherry) reporter was identified in multiple organs and specifically in olfactory microvillous cells, enteric nerves, and importantly in respiratory and GI tuft cells. In the small intestine, cell ablation targeting Gpr64-expressing epithelial cells eliminated tuft cells. Transcriptional analysis of small intestinal Gpr64(mCherry) -positive tuft cells confirmed expression of Gpr64 and the chemo-sensors Sucnr1, Gprc5c, Drd3, and Gpr41/Ffar3. Time-lapse studies of organoids from Trpm5(GFP):Gpr64(mCherry) mice revealed sequential expression of initially Trpm5(GFP) and subsequently also Gpr64(mCherry) 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; however, in the GI tract, GPR64 is remarkably selectively expressed in mature versus young immunoregulatory tuft cells. |
format | Online Article Text |
id | pubmed-8105302 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | Elsevier |
record_format | MEDLINE/PubMed |
spelling | pubmed-81053022021-05-14 Adhesion receptor ADGRG2/GPR64 is in the GI-tract selectively expressed in mature intestinal tuft cells Grunddal, Kaare V. Tonack, Sarah Egerod, Kristoffer L. Thompson, Jonathan James Petersen, Natalia Engelstoft, Maja S. Vagne, Constance Keime, Céline Gradwohl, Gérard Offermanns, Stefan Schwartz, Thue W. Mol Metab Original Article OBJECTIVE: GPR64/ADGRG2 is an orphan Adhesion G protein-coupled receptor (ADGR) known to be mainly expressed in the parathyroid gland and epididymis. This investigation aimed to delineate the cellular expression of GPR64 throughout the body with focus on the gastrointestinal (GI) tract. METHODS: Transgenic Gpr64(mCherry) 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 Gpr64(mCherry)-positive tuft cells was analyzed by quantitative RT-PCR analysis and in situ hybridization. The Gpr64(mCherry) was crossed into the general tuft cell reporter Trpm5(GFP) 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 Gpr64(mCherry) reporter was identified in multiple organs and specifically in olfactory microvillous cells, enteric nerves, and importantly in respiratory and GI tuft cells. In the small intestine, cell ablation targeting Gpr64-expressing epithelial cells eliminated tuft cells. Transcriptional analysis of small intestinal Gpr64(mCherry) -positive tuft cells confirmed expression of Gpr64 and the chemo-sensors Sucnr1, Gprc5c, Drd3, and Gpr41/Ffar3. Time-lapse studies of organoids from Trpm5(GFP):Gpr64(mCherry) mice revealed sequential expression of initially Trpm5(GFP) and subsequently also Gpr64(mCherry) 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; however, in the GI tract, GPR64 is remarkably selectively expressed in mature versus young immunoregulatory tuft cells. Elsevier 2021-04-05 /pmc/articles/PMC8105302/ /pubmed/33831593 http://dx.doi.org/10.1016/j.molmet.2021.101231 Text en © 2021 The Authors https://creativecommons.org/licenses/by/4.0/This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Original Article Grunddal, Kaare V. Tonack, Sarah Egerod, Kristoffer L. Thompson, Jonathan James Petersen, Natalia Engelstoft, Maja S. Vagne, Constance Keime, Céline Gradwohl, Gérard Offermanns, Stefan Schwartz, Thue W. Adhesion receptor ADGRG2/GPR64 is in the GI-tract selectively expressed in mature intestinal tuft cells |
title | Adhesion receptor ADGRG2/GPR64 is in the GI-tract selectively expressed in mature intestinal tuft cells |
title_full | Adhesion receptor ADGRG2/GPR64 is in the GI-tract selectively expressed in mature intestinal tuft cells |
title_fullStr | Adhesion receptor ADGRG2/GPR64 is in the GI-tract selectively expressed in mature intestinal tuft cells |
title_full_unstemmed | Adhesion receptor ADGRG2/GPR64 is in the GI-tract selectively expressed in mature intestinal tuft cells |
title_short | Adhesion receptor ADGRG2/GPR64 is in the GI-tract selectively expressed in mature intestinal tuft cells |
title_sort | adhesion receptor adgrg2/gpr64 is in the gi-tract selectively expressed in mature intestinal tuft cells |
topic | Original Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8105302/ https://www.ncbi.nlm.nih.gov/pubmed/33831593 http://dx.doi.org/10.1016/j.molmet.2021.101231 |
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