Inhibition of noradrenergic signalling in rodent orbitofrontal cortex impairs the updating of goal-directed actions
- CNRS, University of Bordeaux, France
- Institut de Génétique Moléculaire de Montpellier, CNRS, University of Montpellier, France
Figures
Figure 1 with 3 supplements
Initial goal-directed learning does not require NA signalling in the OFC.
(A) Experimental timeline for rats injected with anti-DβH saporin (SAP) or the inactive control (CTL) toxin before (Pre) and after (Post) initial instrumental training and outcome devaluation …
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Figure 1—source data 1
Source files for the quantification of dopamine beta hydroxylase (DβH)-positive fibres (ventral orbitofrontal cortex [VO], lateral orbitofrontal cortex [LO], and Area 32 [A32]) and behavioural data for rats injected with saporin and inactive saporin.
This excel file contains the raw data from the quantification of DβH-positive fibres in the VO, LO, and Area 32 dorsal (A32d) and ventral (A32v). Behavioural data from initial and reversal phases are also supplied.
- https://cdn.elifesciences.org/articles/81623/elife-81623-fig1-data1-v2.xlsx
Figure 1—figure supplement 1
Whole-brain (2.5×) and zoomed-in (20×) photomicrographs showing the volume of dopamine beta hydroxylase (DβH)-positive fibres (%) in the ventral orbitofrontal cortex (VO), lateral orbitofrontal cortex (LO), and Area 32 (A32) of a representative control (CTL) animal and of two saporin (SAP)-treated rats, one with the maximum (MAX) depletion and one with the minimum (MIN) depletion.
Figure 1—figure supplement 2
Quantification of dopamine beta hydroxylase (DβH)-positive fibres in other prefrontal cortex regions.
(A) Regions where dopamine beta hydroxylase (DβH)-positive fibres were quantified (medial orbitofrontal [MO], secondary motor cortex [M2], insula); schematics adapted from of The Rat Brain in …
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Figure 1—figure supplement 2—source data 1
Source files for the quantification of dopamine beta hydroxylase (DβH)-positive fibres (medial orbitofrontal cortex [MO], secondary motor cortex [M2], and insula) for rats injected with saporin and inactive saporin.
This excel file contains the raw data from the quantification of DβH-positive fibres in MO, M2, and insular cortex.
- https://cdn.elifesciences.org/articles/81623/elife-81623-fig1-figsupp2-data1-v2.xlsx
Figure 1—figure supplement 3
Consumption tests performed immediately after the initial (A) and reversal (B) instrumental tests (Ndev: non-devalued; Dev: devalued).
Rats were given access to both food rewards (10 g each) for 10 min. Statistics revealed a within-subjects effect of devaluation for both the initial (F(1,53) = 168.94, p<0.001) and reversal tests (F(…
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Figure 1—figure supplement 3—source data 1
Source files for the consumption tests for rats injected with saporin and inactive saporin.
This excel file contains the raw data from the consumption tests conducted after the initial outcome devaluation test and the reversal outcome devaluation test.
- https://cdn.elifesciences.org/articles/81623/elife-81623-fig1-figsupp3-data1-v2.xlsx
Figure 2 with 2 supplements
Regions where tyrosine hydroxylase (TH, panel A) and dopamine beta hydroxylase (DβH, panel C) positive fibres were quantified (Area 32 dorsal: A32d; Area 32 ventral: A32v; ventral orbitofrontal cortex: VO; lateral orbitofrontal cortex: LO); schematic adapted from Figure 9 of The Rat Brain in Stereotaxic Coordinates (Paxinos and Watson, 2014).
Representative photomicrographs and quantification of fibres in VO, LO, and A32d for TH (B) and DβH immunostaining (D) in each of the three groups. Data are presented as mean + SEM. **p<0.01, ***p<0.…
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Figure 2—source data 1
Source files for the quantification of tyrosine hydroxylase (TH)-positive and dopamine beta hydroxylase (DβH)-positive fibres (ventral orbitofrontal cortex [VO], lateral orbitofrontal cortex [LO], and Area 32 [A32]) for rats injected with 6-hydroxydopamine (6-OHDA) or 6-OHDA+desipramine.
This excel file contains the raw data from the quantification of TH- and DβH-positive fibres in the VO, LO, and A32.
- https://cdn.elifesciences.org/articles/81623/elife-81623-fig2-data1-v2.xlsx
Figure 2—figure supplement 1
Whole-brain (2.5×) and zoomed-in (20×) photomicrographs showing the volume of dopamine beta hydroxylase (DβH)-positive fibres (%) in the ventral orbitofrontal cortex (VO), lateral orbitofrontal cortex (LO), and Area 32 (A32) of a representative control (CTL), 6-OHDA+desipramine (6-OHDA+Desi), and 6-OHDA animal.
Figure 2—figure supplement 2
Quantification of dopamine beta hydroxylase (DβH)-positive fibres in other prefrontal cortex regions.
(A) Regions where dopamine beta hydroxylase (DβH)-positive fibres were quantified (medial orbitofrontal [MO], secondary motor cortex [M2], insula); schematics adapted from of The Rat Brain in …
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Figure 2—figure supplement 2—source data 1
Source files for the quantification of tyrosine hydroxylase (TH)-positive and dopamine beta hydroxylase (DβH)-positive fibres (medial orbitofrontal cortex [MO], secondary motor cortex [M2], and insula) for rats injected with 6-hydroxydopamine (6-OHDA) or 6-OHDA+desipramine.
This excel file contains the raw data from the quantification of TH- and DβH-positive fibres in MO, M2, and insula cortex.
- https://cdn.elifesciences.org/articles/81623/elife-81623-fig2-figsupp2-data1-v2.xlsx
Figure 3 with 2 supplements
NA, but not DA, signalling in the OFC is required to adapt to changes in outcome identity.
(A) Experimental timeline. After the initial instrumental training and outcome devaluation testing, rats were injected in the orbitofrontal cortices (OFC) with either vehicle (CTL n=8; CTL n=8), …
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Figure 3—source data 1
Source files for the behavioural data from the reversal phase for rats injected with 6-hydroxydopamine (6-OHDA) or 6-OHDA+desipramine.
This excel file contains the behavioural data from the reversal training and outcome devaluation test for rats injected with 6-OHDA or 6-OHDA+desipramine and their respective controls.
- https://cdn.elifesciences.org/articles/81623/elife-81623-fig3-data1-v2.xlsx
Figure 3—figure supplement 1
Initial training and test for rats to be injected with 6-OHDA (n=12) and control rats (CTL; n=8).
(A) Training data is presented collapsed across the two actions (A1–O1; A2–O2). There was a main effect of group (F(1,18) = 35.33, p<0.001), training day (F(1,18) = 190.66, p<0.001), and group × day …
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Figure 3—figure supplement 1—source data 1
Source files for the behavioural data from the initial phase for rats injected with 6-hydroxydopamine (6-OHDA).
This excel file contains the behavioural data from the initial training, outcome devaluation test, and the consumption tests (initial and reversal) for rats injected with 6-OHDA and their respective controls.
- https://cdn.elifesciences.org/articles/81623/elife-81623-fig3-figsupp1-data1-v2.xlsx
Figure 3—figure supplement 2
Initial training and test for rats to be injected with 6-OHDA+Desi (n=9) and control rats (CTL; n=8).
(A) Training data is presented collapsed across the two actions (A1–O1; A2–O2). There was a main effect of training day (F(1,15) = 220.45, p<0.001), but no effect of group (F(1,15) = 0.06, p=0.81) …
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Figure 3—figure supplement 2—source data 1
Source files for the behavioural data from the initial phase for rats injected with 6-hydroxydopamine (6-OHDA)+desipramine.
This excel file contains the behavioural data from the initial training, outcome devaluation test, and the consumption tests (initial and reversal) for rats injected with 6-OHDA+desipramine and their respective controls.
- https://cdn.elifesciences.org/articles/81623/elife-81623-fig3-figsupp2-data1-v2.xlsx
Figure 4 with 1 supplement
Histological examination following CAV2-PRS-hM4Di-mCherry injections.Validation of the CAV2-PRS-hM4Di-mCherry construct.
(A) CAV2-PRS-hM4Di-mCherry, a vector bearing a noradrenergic (NA)-specific promoter (PRS) that drives the expression of inhibitory DREADDs tagged with HA (hM4Di), and an mCherry expression cassette …
Figure 4—figure supplement 1
Validation of the CAV2-PRS-hM4Di-mCherry construct.
(A) Representative image showing locus coeruleus (LC) cells projecting to A32 and expressing inhibitory DREADDs (hM4Di, mCherry) and LC cells activated by stress (c-Fos, Alexa 488). The …
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Figure 4—figure supplement 1—source data 1
Source file for the quantification of locus coeruleus (LC) hM4Di/C-Fos-positive cells.
This excel file contains the data from the quantification of hM4Di/c-Fos-positive cells, as well as the percentages of colocalization, for rats injected with CAV2-PRS-hM4Di-mCherry in Area 32 (A32) that underwent the stress procedure.
- https://cdn.elifesciences.org/articles/81623/elife-81623-fig4-figsupp1-data1-v2.xlsx
Figure 5 with 2 supplements
Silencing of LC:vlOFC, but not LC:A32, projections impairs adaptation to changes in the A-O association.
(A) Timeline for rats injected with CAV2-PRS-hM4Di-mCherry in either the orbitofrontal cortices (OFC) or Area 32 (A32). Each rat was injected with either vehicle (-) or DCZ (+) during the reversal …
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Figure 5—source data 1
Source file for the behavioural data from the reversal phase for LC:vlOFC and LC:A32 rats.
This excel file contains the behavioural data from the reversal training and outcome devaluation tests for rats injected with CAV2-PRS-hM4Di-mCherry in the OFC (vlOFC) or Area 32 (A32).
- https://cdn.elifesciences.org/articles/81623/elife-81623-fig5-data1-v2.xlsx
Figure 5—figure supplement 1
Initial training and test for rats injected with CAV2-PRS-hM4Di-mCherry in the orbitofrontal cortex (OFC).
(A) Initial training for rats injected with CAV2-PRS-hM4Di-mCherry in the orbitofrontal cortex (OFC) that would be allocated to group vehicle (Veh; n=12) and group deschloroclozapine (DCZ; n=13) for …
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Figure 5—figure supplement 1—source data 1
Source file for the behavioural data from the initial phase for LC:vlOFC rats.
This excel file contains the behavioural data from the initial training and outcome devaluation test, as well as the consumption tests (initial and reversal) for rats injected with CAV2-PRS-hM4Di-mCherry in the orbitofrontal cortex (OFC).
- https://cdn.elifesciences.org/articles/81623/elife-81623-fig5-figsupp1-data1-v2.xlsx
Figure 5—figure supplement 2
Initial training and test for rats injected with CAV2-PRS-hM4Di-mCherry in area 32 (A32).
(A) Initial training for rats injected with CAV2-PRS-hM4Di-mCherry in Area 32 (A32) that would be allocated to group vehicle (Veh; n=8) and group deschloroclozapine (DCZ; n=9) for the subsequent …
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Figure 5—figure supplement 2—source data 1
Source file for the behavioural data from the initial phase for LC:A32 rats.
This excel file contains the behavioural data from the initial training and outcome devaluation test, as well as the consumption tests (initial and reversal) for rats injected with CAV2-PRS-hM4Di-mCherry in A32.
- https://cdn.elifesciences.org/articles/81623/elife-81623-fig5-figsupp2-data1-v2.xlsx
Tables
Key resources table
| Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| Antibody | Anti-DβH (mouse monoclonal) | Merck Millipore | Cat# MAB308 | 1:1000 |
| Antibody | Anti-TH (mouse monoclonal) | Merck Millipore | Cat# MAB318 | 1:2000 |
| Antibody | Anti-RFP (rabbit polyclonal) | MBL International Corporation | Cat# PM005 | 1:2000 |
| Antibody | Anti-HA (rabbit monoclonal) | Cell Signaling Technology | Cat# C29F4 (#3724) | 1:1000 |
| Antibody | Anti-c-Fos (rabbit monoclonal) | Cell Signaling Technology | Cat# 9F6 (#2250) | 1:1000 |
| Antibody | Anti-mouse biotin-conjugated (goat polyclonal) | Jackson ImmunoResearch | Cat# 115-065-062 | 1:1000 |
| Antibody | Anti-rabbit biotin-conjugated (goat polyclonal) | Jackson ImmunoResearch | Cat# 111-065-003 | 1:1000 |
| Antibody | Anti-mouse FITC-conjugated (goat polyclonal) | Jackson ImmunoResearch | Cat# 115-095-003 | 1:400 |
| Antibody | Anti-rabbit TRITC-conjugated (goat polyclonal) | Jackson ImmunoResearch | Cat# 111-025-003 | 1:200 |
| Antibody | Anti-rabbit AF488-conjugated (goat polyclonal) | Jackson ImmunoResearch | Cat# 111-545-003 | 1:1000 |
| Chemical compound, drug | Streptavidin-Alexa 488 | Thermo Fisher Scientific | Cat# S11223 | 1:500 |
| Chemical compound, drug | Anti-DβH saporin | Advanced Targeting Solutions | Cat# KIT-03 | |
| Chemical compound, drug | Mouse IgG saporin (inactive) | Advanced Targeting Solutions | Cat# IT-18 sold as KIT-03 | |
| Chemical compound, drug | Diaminobenzidine (DAB) | Sigma-Aldrich | Cat# D5905 | |
| Chemical compound, drug | Deschloroclozapine (DCZ) | MedChemExpress | Cat# HY-42110 | Injectable volume 0.1 mg/kg |
| Chemical compound, drug | 6-OHDA hydrochloride | Sigma-Aldrich | Cat# H4381 | |
| Chemical compound, drug | Desipramine | Sigma-Aldrich | Cat# D3900 | |
| Commercial assay or kit | Avidin-biotin-peroxydase (ABC kit) | Thermo Fisher Scientific | Cat# 32020 | |
| Transfected construct (human) | CAV2 PRS HA-hM4Di E1 hSyn mCherry E3 | https://plateau-igmm.pvm.cnrs.fr/?vector=cav-prs-ha-hm4di | Titre 3.5×1012 pp/mL |
