Expression Mapping and Functional Analysis of Orphan G-Protein-Coupled Receptor GPR158 in the Adult Mouse Brain Using a GPR158 Transgenic Mouse

Jinlong Chang; Ze Song; Shoupeng Wei; Yunxia Zhou; Jun Ju; Peijia Yao; Youheng Jiang; Hui Jin; Xinjin Chi; Ningning Li

doi:10.3390/biom13030479

Expression Mapping and Functional Analysis of Orphan G-Protein-Coupled Receptor GPR158 in the Adult Mouse Brain Using a GPR158 Transgenic Mouse

Biomolecules. 2023 Mar 5;13(3):479. doi: 10.3390/biom13030479.

Authors

Jinlong Chang¹, Ze Song², Shoupeng Wei¹, Yunxia Zhou¹, Jun Ju¹, Peijia Yao¹, Youheng Jiang¹, Hui Jin¹, Xinjin Chi³, Ningning Li^{1

4

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Affiliations

¹ Tomas Lindahl Nobel Laureate Laboratory, The Seventh Affiliated Hospital, Sun Yat-Sen University, Shenzhen 518107, China.
² The Clinical Oncology Department, The Seventh Affiliated Hospital, Sun Yat-Sen University, Shenzhen 518107, China.
³ Department of Anesthesiology, The Seventh Affiliated Hospital, Sun Yat-Sen University, Shenzhen 518107, China.
⁴ China-UK Institute for Frontier Science, Shenzhen 518107, China.
⁵ The Fifth People's Hospital of Datong City, Datong 038300, China.

Abstract

Aberrant expression of G-protein-coupled receptor 158 (GPR158) has been reported to be inextricably linked to a variety of diseases affecting the central nervous system, including Alzheimer's disease (AD), depression, intraocular pressure, and glioma, but the underlying mechanism remains elusive due to a lack of biological and pharmacological tools to elaborate its preferential cellular distribution and molecular interaction network. To assess the cellular localization, expression, and function of GPR158, we generated an epitope-tagged GPR158 mouse model (GPR158^Tag) that exhibited normal motor, cognitive, and social behavior, no deficiencies in social memory, and no anxiety-like behavior compared to C57BL/6J control mice at P60. Using immunofluorescence, we found that GPR158⁺ cells were distributed in several brain regions including the cerebral cortex, hippocampus, cerebellum, and caudate putamen. Next, using the cerebral cortex of the adult GPR158^Tag mice as a representative region, we found that GPR158 was only expressed in neurons, and not in microglia, oligodendrocytes, or astrocytes. Remarkably, the majority of GPR158 was enriched in Camk2a⁺ neurons whilst limited expression was found in PV⁺ interneurons. Concomitant 3D co-localization analysis revealed that GPR158 was mainly distributed in the postsynaptic membrane, but with a small portion in the presynaptic membrane. Lastly, via mass spectrometry analysis, we identified proteins that may interact with GPR158, and the relevant enrichment pathways were consistent with the immunofluorescence findings. RNA-seq analysis of the cerebral cortex of the GPR158^-/- mice showed that GPR158 and its putative interacting proteins are involved in the chloride channel complex and synaptic vesicle membrane composition. Using these GPR158^Tag mice, we were able to accurately label GPR158 and uncover its fundamental function in synaptic vesicle function and memory. Thus, this model will be a useful tool for subsequent biological, pharmacological, and electrophysiological studies related to GPR158.

Keywords: 3D co-location; GPR158; brain regions; cell types; cerebral cortex; protein–protein interaction; subcellular localization.

Publication types

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

MeSH terms

Animals
Brain* / metabolism
Cell Communication
Mice
Mice, Inbred C57BL
Mice, Transgenic
Receptors, G-Protein-Coupled* / genetics
Receptors, G-Protein-Coupled* / metabolism

Substances

Receptors, G-Protein-Coupled
GPR158 protein, mouse

Grants and funding

This study was supported by grants from the Hundred Talents Program of Sun Yat-sen University (392007, NL), National Natural Science Foundation of China (81874176 and 82072766, NL; 82201698, SW), Shenzhen Sanming Project of Medicine (SZSM201911003, NL), Shenzhen Science, Technology and Innovation Commission (SZSTI) Basic Research Program (JCYJ20190809154411427 and JCYJ20220530145008018, NL; No. JCYJ20210324134800002, SW), and Guangdong Basic and Applied Basic Research Foundation (2020A1515110161, JJ; 2021A1515110891, SW).