Identification of G Protein-Coupled Receptors (GPCRs) in Primary Cilia and Their Possible Involvement in Body Weight Control

Yoshihiro Omori; Taro Chaya; Satoyo Yoshida; Shoichi Irie; Toshinori Tsujii; Takahisa Furukawa

doi:10.1371/journal.pone.0128422

Identification of G Protein-Coupled Receptors (GPCRs) in Primary Cilia and Their Possible Involvement in Body Weight Control

PLoS One. 2015 Jun 8;10(6):e0128422. doi: 10.1371/journal.pone.0128422. eCollection 2015.

Authors

Yoshihiro Omori¹, Taro Chaya¹, Satoyo Yoshida², Shoichi Irie², Toshinori Tsujii², Takahisa Furukawa²

Affiliations

¹ Laboratory for Molecular and Developmental Biology, Institute for Protein Research, Osaka University, 3-2 Yamadaoka, Suita, Osaka, Japan; JST, PRESTO, 3-2 Yamadaoka, Suita, Osaka, Japan.
² Laboratory for Molecular and Developmental Biology, Institute for Protein Research, Osaka University, 3-2 Yamadaoka, Suita, Osaka, Japan.

Abstract

Primary cilia are sensory organelles that harbor various receptors such as G protein-coupled receptors (GPCRs). We analyzed subcellular localization of 138 non-odorant GPCRs. We transfected GPCR expression vectors into NIH3T3 cells, induced ciliogenesis by serum starvation, and observed subcellular localization of GPCRs by immunofluorescent staining. We found that several GPCRs whose ligands are involved in feeding behavior, including prolactin-releasing hormone receptor (PRLHR), neuropeptide FF receptor 1 (NPFFR1), and neuromedin U receptor 1 (NMUR1), localized to the primary cilia. In addition, we found that a short form of dopamine receptor D2 (DRD2S) is efficiently transported to the primary cilia, while a long form of dopamine receptor D2 (DRD2L) is rarely transported to the primary cilia. Using an anti-Prlhr antibody, we found that Prlhr localized to the cilia on the surface of the third ventricle in the vicinity of the hypothalamic periventricular nucleus. We generated the Npy2r-Cre transgenic mouse line in which Cre-recombinase is expressed under the control of the promoter of Npy2r encoding a ciliary GPCR. By mating Npy2r-Cre mice with Ift80 flox mice, we generated Ift80 conditional knockout (CKO) mice in which Npy2r-positive cilia were diminished in number. We found that Ift80 CKO mice exhibited a body weight increase. Our results suggest that Npy2r-positive cilia are important for body weight control.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Body Weight*
Cilia / metabolism*
Hypothalamus / metabolism
Integrases / metabolism
Mice
Mice, Knockout
NIH 3T3 Cells
Protein Isoforms / metabolism
Protein Transport
Receptors, Dopamine D2 / metabolism
Receptors, G-Protein-Coupled / metabolism*
Subcellular Fractions / metabolism
Weight Gain

Substances

Prlhr protein, mouse
Protein Isoforms
Receptors, Dopamine D2
Receptors, G-Protein-Coupled
Cre recombinase
Integrases

Grants and funding

This work was supported by PRESTO (#4118 for YO) from Japan Science and Technology Agency (http://www.jst.go.jp/), Grants-in-Aid for Scientific Research on Innovative Areas (#25113519, # 23113728 for YO), Grant-in-Aid for Scientific Research (B) (#15H04669 for TF, # 25293070 for YO) from Japan Society for the Promotion of Science (http://www.jsps.go.jp/), The Takeda Science Foundation (for YO and TF) (http://www.takeda-sci.or.jp/), The Uehara Memorial Foundation (for YO) (http://www.ueharazaidan.or.jp/), Research Foundation for Opto-Science and Technology (for TF) (http://www.refost-hq.jp/gaiyo.html), Suzuken Memorial Foundation (for YO) (http://www.suzukenzaidan.or.jp/), Naito Foundation (#220 for YO) (https://www.naito-f.or.jp/), Hyogo Science and Technology Association (for TF) (http://www.hyogosta.jp/), The Osaka Community Foundation (for TF) (http://www.osaka-community.or.jp/). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.