Orphan receptor GPR158 controls stress-induced depression
- The Scripps Research Institute, United States
- Max Planck Florida Institute for Neuroscience, United States
- University of Brescia, Italy
- University of South Dakota, United States
- University of Iowa, United States
- Max Planck Institute of Neurobiology, Germany
Figures
Figure 1 with 2 supplements
Stress upregulates GPR158 expression in the mPFC.
(A) RNA sequencing results from the whole mouse brain showing the top 30 orphan receptors. (B) Mass spectrometry analysis of the orphan receptors expressed in the PFC of mice. (C) Representative …
Figure 1—figure supplement 1
GPR158 is among the most expressed GPCR in the PFC.
(A) A proteomic analysis of the GPCR expression pattern in a membrane-enriched fraction of mouse mPFC identified 48 GPCRs with GPR158 being one of the most expressed. (B) Representative western …
Figure 1—figure supplement 2
GPR158 expression in several neuronal populations.
(A) In situ hybridization specificity control. In situ hybridization of mouse GPR158 (left) and of the bacterial gene DapB (right) in the mPFC (scale bar = 50 µm). (B) Double in situ hybridization …
Figure 2
Chronic stress regulates GPR158 levels in glutamatergic neurons of the layer 2/3 of the mPFC.
(A) Representative image of in situ hybridization using a probe against GPR158 (yellow) and co-immunostaining with antibodies against NeuN (red) and GAD67 (green) in the layer 2/3 of mPFC (nuclei …
Figure 3
GPR158 is increased in the mouse mPFC by a corticosterone-mediated mechanism.
(A) GPR158 protein levels are increased in the mPFC of mice chronically treated with corticosterone (n = 9–10 mice/group, Student’s t test). Top panel depicts corticosterone paradigm. Mice treated …
Figure 4 with 1 supplement
GPR158 overexpression in mouse mPFC and in dlPFC of MDD patient.
(A) Scheme of Punisher-Adenovirus expressing GPR158 (GPR158-Ad) and EGFP (EGFP-Ad) injected bilaterally into the mPFC of C57Bl/6J mice. (B) Representative viral expression in the mPFC (right panel). …
Figure 4—figure supplement 1
GPR158 expression levels in patients affected by MDD.
(A) Western blots of GPR158 expression in the dorsolateral prefrontal cortex (dlPFC) of patients with major depressive disorder (MDD) or health unaffected controls (Crtl). (B) Western blot of human …
Figure 5 with 2 supplements
Ablation of GPR158 in mice results in an antidepressant-like phenotype and resiliency to chronic stress.
(A) Gpr158−/− mice display a lower SIH response (T2–T1) compared to Gpr158+/+ mice without affecting their baseline temperature (right panel) (n = 15–17/genotype, Student’s t test, *p<0.05). (B) Gpr1…
Figure 5—figure supplement 1
Depressive- and anxiety-related behaviors are altered in both male and female Gpr158−/− mice.
(A) Locomotor activity in 10 min bins (n = 5–6 mice/genotype). No significant difference between genotypes were found. (B) Effects of Gpr158−/− and Grp158+/+ mice on the accelerated rotarod test …
Figure 5—figure supplement 2
Viral-mediated expression of GPR158 in mPFC of Gpr158−/− mice rescues the antidepressant-like phenotype.
(A) mPFC of Gpr158−/− mice was injected bilaterally with Adenovirus expressing either EGFP or GPR158. Western blot showing the expression of GPR158 only in mice injected with GPR158-Ad (right …
Figure 6
Gpr158−/− mice are resilient to unpredictable chronic mild stress (UCMS).
Individual behavioral measures were integrated to calculate the emotionality score for each animal. Behavioral analysis of Gpr158+/+ and Gpr158−/− mice subjected to UCMS in the (A) marble burying …
Figure 7 with 1 supplement
Synaptic adaptations in layer 2/3 neurons of mPFC induced by GPR158 loss.
(A) Representative images and summarized data showing that the spine density was increased in Gpr158−/− neurons (n = 7–5). (B) Representative traces and summarized data showing that the frequency (C)…
Figure 7—figure supplement 1
AMPA receptors are involved in the synaptic adaptations of Gpr158−/− mice.
(A) Representative current traces are shown and Paired pulse ratio comparison showing no significant differences in Gpr158+/+ and Gpr158−/− mice (n = 11–9 neurons/3 mice). (B) AMPAR and (C) NMDAR …
Figure 8
GPR158 modulates BDNF protein levels in the mPFC.
Representative western blots (A) and quantification (B) of the protein levels and phosphorylation state of the indicated proteins in the mPFC of Gpr158+/+ and Gpr158−/− mice (n = 4–6). (C) Western …
Schema 1
Timeline for behavioral evaluation in stress-induced depression models.
https://doi.org/10.7554/eLife.33273.016Tables
Key resources table
| Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| Antibody | GAPDH | Millipore | Cat# AB2302 | (1:30000) |
| Antibody | GluA1 | Abcam | Cat# ab76321 | (1:1000) |
| Antibody | p831 GluA1 | Millipore | Cat# 04–823 | (1:1000) |
| Antibody | p845 GluA1 | Cell Signaling | Cat# 8084 | (1:1000) |
| Antibody | GluA2 | Millipore | Cat# MAB N71 | (1:1000) |
| Antibody | NR1 | Zymed | Cat# 32–500 | (1:1000) |
| Antibody | p897 NR1 | Millipore | Cat# 06–641 | (1:1000) |
| Antibody | NR2B | Millipore | Cat# AB1557P | (1:1000) |
| Antibody | Rabbit anti-GPR158CT | PMID:25792749 | N/A | (1:1000) |
| Antibody | Rabbit anti-Gβ5 | PMID:15632198 | N/A | (1:1000) |
| Antibody | Rabbit anti-Gβ1 | gift from Dr. Willardson PMID:15485848 | N/A | 1:6000 |
| Antibody | GPR34 | ThermoFisher Scientific | Cat# PA5-45717 | (1:1000) |
| Antibody | GPR162 | Aviva Systems Biology | Cat# ARP68400 | (1:1000) |
| Antibody | GPRC5B | Aviva Systems Biology | Cat# ARP59799 | (1:1000) |
| Antibody | AKT | Cell Signaling | Cat# 9272 | (1:1000) |
| Antibody | p473 AKT | Cell Signaling | Cat# 4060 | (1:1000) |
| Antibody | mTOR | Cell Signaling | Cat# 2983 | (1:1000) |
| Antibody | S6 Kinase (P70) | Cell Signaling | Cat# 9202 | (1:1000) |
| Antibody | ERK1/2 | Cell Signaling | Cat# 9102 | (1:1000) |
| Antibody | GSK-3β | Cell Signaling | Cat# 9315 | (1:1000) |
| Antibody | p9 GSK3α/β | Cell Signaling | Cat# 9331 | (1:1000) |
| Antibody | BDNF | Abcam | Cat# ab108319 | (1:1000) |
| Antibody | TrkB | Cell Signaling | Cat# 4603 | (1:1000) |
| Antibody | MeCP2 | Millipore | Cat# 07–013 | (1:1000) |
| Antibody | Gαo | Cell Signaling | Cat# 3975 | (1:1000) |
| Antibody | PLCβ2 | Santa Cruz | Cat# sc206 | (1:200) |
| Antibody | PP2Acat | Assay Biotech | Cat# B0555 | (1:1000) |
| Antibody | PP2Areg | Cell Signaling | Cat# 2041 | (1:1000) |
| Antibody | SYP | Assay Biotech | Cat# C0333 | (1:1000) |
| Antibody | PSD95 | Cell Signaling | Cat# 3450 | (1:1000) |
| Antibody | eEF2 | Cell Signaling | Cat# 2332 | (1:1000) |
| Antibody | p56 eEF2 | Cell Signaling | Cat# 2331 | (1:1000) |
| Antibody | NeuN | Abcam | Cat# 104225 | (1:1000) |
| Antibody | GAD67 | Millipore | Cat# MAB5406 | (1:1000) |
| Genetic reagent (adenovirus) | HdAd-23E4-Pun-GPR158-syn-EGFP | This Paper | N/A | |
| Genetic reagent (adenovirus) | HdAd-23E4-Pun-Syn-EGFP | This Paper | N/A | |
| Biological sample (brain tissue) | Adult human brain tissue (healthy and MDD samples) | NIH NeuroBioBank (University of Pittsburg, University of Miami Brain Endowment Bank and the Human Brain and Spinal Fluid Resource Center) | 13138, 13054, 13082, 5289, 002, 006, 5293, AN04642, AN04917, AN07465, AN14368, AN15392, AN01077, AN05364, 13006, 13011, 13022, 13047, 13075, 13086, 13145, 13057, 13051, 13181, 535, 556, 711 | |
| Chemical compound, drug | Corticosterone | Sigma-Aldridge | Cat# C2505 | |
| Chemical compound, drug | RU-486 | Sigma-Aldridge | Cat# M8046 | |
| Chemical compound, drug | Dexamethasone | Tocris | Cat# D4902 | |
| Chemical compound, drug | NBQX | Tocris | Cat# 10441R | |
| commercial assay or kit | QuantiGene ViewRNA ISH Tissue 2-Plex Assay | Affymetrix | Cat# QVT0012 | |
| commercial assay or kit | Gpr158 NM_001004761; ViewRNA TYPE 1 Probe | Affymetrix | Cat# VB1-11518 | |
| commercial assay or kit | Slc17a7 NM _182993; ViewRNA TYPE 6 Probe | Affymetrix | Cat# VB6-14162 | |
| commercial assay or kit | Gad1 NM_008077; ViewRNA TYPE 6 Probe | Affymetrix | Cat# VB6-12632 | |
| commercial assay or kit | Pvalb NM_013645; ViewRNA TYPE 6 Probe | Affymetrix | Cat# VB6-13220 | |
| commercial assay or kit | Sst NM_009215; ViewRNA TYPE 6 Probe | Affymetrix | Cat# VB6-13754 | |
| commercial assay or kit | DapB NC_000913; ViewRNA TYPE 1 Probe | Affymetrix | Cat# VF1-10272 | |
| commercial assay or kit | Direct cAMP ELISA kit | ENZO Life Sciences | Cat# ADI-900–066 | |
| strain, strain background (mus musculus) | GPR158 KO | PMID:25792749 | N/A | |
| software, algorithm | ImageJ | Wayne Rasband, NIH, USA | SCR_003070 | |
| software, algorithm | Zen2.1 SP1 (Black) | Carl Zeiss | SCR_013672 | |
| software, algorithm | Prism6 | GraphPad Software | SCR_002798 | |
| software, algorithm | Neurolucida | MBF Bioscience |
Additional files
-
Supplementary file 1
G protein Coupled Receptors identified by mass-spectrometry in the PFC.
- https://doi.org/10.7554/eLife.33273.017
-
Transparent reporting form
- https://doi.org/10.7554/eLife.33273.018
