An endogenous GLP-1 circuit engages VTA GABA neurons to regulate mesolimbic dopamine neurons and attenuate cocaine seeking
Data files
Jan 13, 2025 version files 168.25 KB
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Auxiliary_Supplementary_Data_File_FINAL.xlsx
154.34 KB
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README.md
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Abstract
Recent studies show that systemic administration of a glucagon-like peptide-1 receptor (GLP-1R) agonist is sufficient to attenuate the reinstatement of cocaine-seeking behavior, an animal model of relapse. However, the neural mechanisms mediating these effects and the role of endogenous central GLP-1 signaling in cocaine seeking remain unknown. Here, we show that voluntary cocaine taking decreased plasma GLP-1 levels in rats and that chemogenetic activation of GLP-1-producing neurons in the nucleus tractus solitarius (NTS) that project to the ventral tegmental area (VTA) decreased cocaine reinstatement. Single nuclei transcriptomics and FISH studies revealed GLP-1Rs are expressed primarily on GABA neurons in the VTA. Using in vivo fiber photometry, we found that the efficacy of a systemic GLP-1R agonist to attenuate cocaine seeking was associated with increased activity of VTA GABA neurons and decreased activity of VTA dopamine neurons. Together, these findings suggest that targeting central GLP-1 circuits may be an effective strategy toward reducing cocaine relapse and highlight a novel functional role of GABAergic GLP-1R-expressing midbrain neurons in drug seeking.
README
An endogenous GLP-1 circuit engages VTA GABA neurons to regulate mesolimbic dopamine neurons and attenuate cocaine seeking
We have uploaded all the data associated with this manuscript on Dryad (DOI:10.5061/dryad.djh9w0w8v). Single cell RNA-sequencing data sets can be found at GSE284465.
Description of the data and file structure
All datasets from this manuscript are compiled in the file (Auxiliary Supplementary Data File.CSV). Each sheet in the file includes the data from each of figure panels. Below details a summary of the data and variables present on each sheet of the .CSV file.
Figure 1B: GLP-1 blood plasma levels on day 21 of cocaine self-admninstration
- Animal ID: Rat subject ID name
- cocaine: animals that self-admnistered cocaine (even animal IDs)
- yoked-saline: yoked-saline control animals (odd animal IDs); yoked-saline animals are paired to the even number cocaine animal following (i.e. cocaine 2's yoked-saline control is 1)
- Blood plasma: Value of GLP-1 blood plasma expression in (pg/ml)
Figure 1C: GLP-1 blood plasma levels on day 1 of extinction
Same variables as 1B
Figure 1D: GLP-1 blood plasma levels on day 7 of extinction
Same variables as 1B
Figure 2D
- virus: virus injected into the NTS (either hM3D(Gq) or mCherry control)
- % cFOS + Cherry/total mCherry: (# of cells co-expressing cfos and mCherry) / (total number of mCherry expressing cells)*100%
Figure 2G: Lever responses during reinstatement for hM3D(Gq) injected rats self-admnistering cocaine
- Animal ID: Rat subject ID name
- Treatment: Dose of CNO administered prior to cocaine prime (e.g. 0 mg/kg; 0.1 mg/kg; or 1.0 mg/kg)
- Lever: # of active lever or inactive lever responses during reinstatement test session
Figure 2H: Cummulative chow intake (in grams) following reinstatement tests for hM3D(Gq) injected rats
- Animal ID: Rat subject ID name
- Treatment: Dose of CNO administered prior to cocaine prime (e.g. 0 mg/kg or 1.0 mg/kg)
- Time: Time in hours following end of reinstatement test session
Figure 2I: 24 hour bodyweight change (in grams) following reinstatement tests for hM3D(Gq) injected rats
Bodyweight change is reported in grams per 24 hours
- Animal ID: Rat subject ID name
- Treatment: Dose of CNO administered prior to cocaine prime (e.g. 0 mg/kg or 1.0 mg/kg)
Figure 2J: Lever responses during reinstatement for mCherry injected rats self-admnistering cocaine
- Animal ID: Rat subject ID name
- Treatment: Dose of CNO administered prior to cocaine prime (e.g. 0 mg/kg or 1.0 mg/kg)
- Lever: # of active lever or inactive lever responses during reinstatement test session
Figure 2K: Cummulative chow intake (in grams) following reinstatement tests for mCherry injected rats
- Animal ID: Rat subject ID name
- Treatment: Dose of CNO administered prior to cocaine prime (e.g. 0 mg/kg or 1.0 mg/kg)
- Time: Time in hours following end of reinstatement test session
Figure 2L: 24 hour bodyweight change (in grams) following reinstatement tests for mCherry injected rats
Bodyweight change is reported in grams per 24 hours
- Animal ID: Rat subject ID name
- Treatment: Dose of CNO administered prior to cocaine prime (e.g. 0 mg/kg or 1.0 mg/kg)
Figure 3E: Active lever responses for new cohort of cocaine self-administration hM3D(Gq) injected rats during reinstatement
Gq_1b to Gq_8b refers to the animal ID
- VTA treatment: Dose of Ex-9 administered prior to CNO treatment (e.g. 0 ug/100nl or 10 ug/100nl)
- IP Treatment: Dose of CNO administered prior to cocaine prime (e.g. 0 mg/kg or 1.0 mg/kg)
Figure 3F: Inactive lever responses for new cohort of cocaine self-administration hM3D(Gq) injected rats during reinstatement
Gq_1b to Gq_8b refers to the animal ID
- VTA treatment: Dose of Ex-9 administered prior to CNO treatment (e.g. 0 ug/100nl or 10 ug/100nl)
- IP Treatment: Dose of CNO administered prior to cocaine prime (e.g. 0 mg/kg or 1.0 mg/kg)
Figure 4B,4C, and 4D: Cells expressing glp1r RNA transcripts in the VTA
- Animal ID: Rat subject ID name
- AP (mm): Anterior/posterior position of slice based off rat brain atlas according to Paxinos and Western (mm)
- Brain section: Location of brain section on slide
- GLP-1R: # of dapi+ cells expressing glp1r
- GLP-1R+/GAD+: # of dapi+ cells expressing glp1r and gad
- GLP-1R+/TH+: # of dapi+ cells expressing glp1r and th
Figure 5C: Lever responses during reinstatement for GCaMP8f injected Gad-cre rats self-administering cocaine
VTA1G-VTA5G refers to animal ID ("VTA1_GAD")
- Treatment: Dose of Ex-4 administered prior to cocaine prime (0 ug/kg Ex-4 or 2.0 ug/kg)
- Active Lever Presses: # of active lever respones during reinstatement test session
- Inactive Lever Presses: # of inactive lever responses during reinstatement test session
Figure 5D: GCaMP trace during active lever press in Gad_cre rats
Exendin-4 (0.0 ug/kg) + Cocaine (10mg/kg) and Exendin-4 (2.0 ug/kg) + Cocaine (10mg/kg) refer to reinstatement treatment condition under which the photometry data was recorded
- timestamp: time relative to active lever press (0 seconds) for each z-scored GCaMP vlaue
- VTA#G_al#: Animal ID # and active lever press number (e.g. VTA1G_al1 -> Animal VTA1G active lever press 1)
Figure 5E: Net area under Gad_cre GCaMP8f traces associated with active lever press (analysis of traces in 5D)
Exendin-4 (0.0 ug/kg) + Cocaine (10mg/kg) and Exendin-4 (2.0 ug/kg) + Cocaine (10mg/kg) refer to reinstatement treatment condition under which the photometry data was recorded
- VTA#G_al#: Animal ID # and active lever press number (e.g. VTA1G_al1 -> Animal VTA1G active lever press 1)
- time bucket: data points between which area under the curve is being calculated. "Pre (-5 to 0s)" refers to 5 seconds before active lever press and "Post (0 to 5s)" refers to 5 seconds after active lever press
Figure 5F: Maximum Gad_cre GCaMP8f z-score associated with active lever press (analysis of traces in 5D)
Exendin-4 (0.0 ug/kg) + Cocaine (10mg/kg) and Exendin-4 (2.0 ug/kg) + Cocaine (10mg/kg) refer to reinstatement treatment condition under which the photometry data was recorded
- VTA#G_al#: Animal ID # and active lever press number (e.g. VTA1G_al1 -> Animal VTA1G active lever press 1)
- time bucket: data points between which maximum z-score is being calculated. "Pre (-5 to 0s)" refers to 5 seconds before active lever press and "Post (0 to 5s)" refers to 5 seconds after active lever press
Figure 6C: Lever responses during reinstatement for GCaMP8f injected Th-cre rats
VTA1T-VTA5T refers to animal ID ("VTA1_TH")
Same variables as Figure 5C
Figure 6D: GCaMP trace during active lever press in Th_cre rats
Same variables as
- VTA#T_al#: Animal ID # and active lever press number (e.g. VTA1T_al1 -> Animal VTA1T active lever press 1)
Figure 6E: Net area under Th_cre GCaMP8f traces associated with active lever press (analysis of traces in 6D)
Same variables as 5E
- VTA#T_al#: Animal ID # and active lever press number (e.g. VTA1T_al1 -> Animal VTA1T active lever press 1)
Figure 6F: Maximum Th_cre GCaMP8f z-score associated with active lever press (analysis of traces in 6D)
Same variables as 5F
- VTA#T_al#: Animal ID # and active lever press number (e.g. VTA1T_al1 -> Animal VTA1T active lever press 1)
Figure 7A: Z-scored traces for Th_cre and Gad_cre rats under Exendin-4 (0.0 ug/kg) + Cocaine (10mg/kg) treatment condition
- time bucket (seconds): refers to bucketed second of time for each corresponding data point
- TH-Cre Rats: refers to th_cre rats injected with GCaMP8f
- GAD-Cre Rats: refers to gad_cre rats injected with GCaMP8f
Figure 7C: Z-scored traces for Th_cre and Gad_cre rats under Exendin-4 (2.0 ug/kg) + Cocaine (10mg/kg) treatment condition
same variables as 7A
Figure S1A: Active lever responses for cocaine and yoked-saline animals corresponding to cocaine self-administration day 21 (figure 1b)
- Animal ID: Rat subject ID name
- cocaine: animals that self-administered cocaine (even animal IDs)
- yoked-saline: yoked-saline control animals (odd animal IDs); yoked-saline animals are paired to the even number cocaine animal following (i.e. cocaine 2's yoked-saline control is 1)
- active lever responses: # of active lever responses
Figure S1B: Active lever responses for cocaine and yoked-saline animals corresponding to extinction day 1 (figure 1c)
same variables as S1A
Figure S1C: Active lever responses for cocaine and yoked-saline animals corresponding to extinction day 7 (figure 1D)
same variables as S1A
Figure S1D: Intravenous infusions for cocaine and yoked-saline animals corresponding to cocaine self-administration day 21 (figure 1b)
- Animal ID: Rat subject ID name
- cocaine: animals that self-administered cocaine (even animal IDs)
- yoked-saline: yoked-saline control animals (odd animal IDs); yoked-saline animals are paired to the even number cocaine animal following (i.e. cocaine 2's yoked-saline control is 1)
- infusions: total # of IV infusions
Figure S1E: Intravenous infusions for cocaine and yoked-saline animals corresponding to cocaine day 1 (figure 1c)
same variables as S1D
Figure S1F: Intravenous infusions for cocaine and yoked-saline animals corresponding to cocaine day 7 (figure 1c)
same variables as S1D
Figure S3A: GCaMP trace during inactive lever press in Gad_cre rats
Exendin-4 (0.0 ug/kg) + Cocaine (10mg/kg) and Exendin-4 (2.0 ug/kg) + Cocaine (10mg/kg) refer to reinstatement treatment condition under which the photometry data was recorded
- timestamp: time relative to inactive lever press (0 seconds) for each z-scored GCaMP value
- VTA#G_ial#: Animal ID # and inactive lever press number (e.g. VTA1G_ial1 -> Animal VTA1G inactive lever press 1)
Figure S3B: Net area under Gad_cre GCaMP8f traces associated with inactive lever press (Analysis of traces in S3A)
Exendin-4 (0.0 ug/kg) + Cocaine (10mg/kg) and Exendin-4 (2.0 ug/kg) + Cocaine (10mg/kg) refer to reinstatement treatment condition under which the photometry data was recorded
- VTA#G_al#: Animal ID # and inactive lever press number (e.g. VTA1G_al1 -> Animal VTA1G inactive lever press 1)
- time bucket: data points between which area under the curve is being calculated. "Pre (-5 to 0s)" refers to 5 seconds before inactive lever press and "Post (0 to 5s)" refers to 5 seconds after active lever press
Figure S3C: Maximum Gad_cre GCaMP8f z-score associated with inactive lever press (analysis of traces in S3A)
Exendin-4 (0.0 ug/kg) + Cocaine (10mg/kg) and Exendin-4 (2.0 ug/kg) + Cocaine (10mg/kg) refer to reinstatement treatment condition under which the photometry data was recorded
- VTA#G_al#: Animal ID # and inactive lever press number (e.g. VTA1G_al1 -> Animal VTA1G inactive lever press 1)
- time bucket: data points between which area under the curve is being calculated. "Pre (-5 to 0s)" refers to 5 seconds before inactive lever press and "Post (0 to 5s)" refers to 5 seconds after active lever press
Figure S4A: Lever responses during reinstatement for GCaMP8f injected Gad-cre rats self-administering saline
VTA1GS-VTA3GS refers to animal ID ("VTA1_GADSaline")
Same as variables in 5C
Figure S4B: GCaMP trace during active lever press in Gad_cre saline rats
Same as variables in 5D
Figure S4C: Net area under GAD_cre GCaMP8f traces associated with active lever press (analysis of traces in S4B)
Same variables as 5E
Figure S4D: Maximum Gad_cre GCaMP8f z-score associated with active lever press (analysis of traces in S4B)
Same variables as 5F
Figure S5A: Lever responses during reinstatement for Gad-cre rats infused with hM3D(Gq) into the VTA
- Treatment: Dose of CNO administered prior to cocaine prime (0 mg/kg CNO, 0.1 mg/kg CNO, or 1.0 mg/kg CNO)
- Active Lever Responses: # of active lever respones during reinstatement test session
- Inactive Lever Responses: # of inactive lever responses during reinstatement test session
Figure S6A: GCaMP trace during inactive lever press in Th_cre rats
same variables as S3A
- VTA#T_ial#: Animal ID # and inactive lever press number (e.g. VTA1G_ial1 -> Animal VTA1T inactive lever press 1)
Figure S6B: Net area under Th_cre GCaMP8f traces associated with inactive lever press (Analysis of traces in S6A)
same variables as S3B
- VTA#T_ial#: Animal ID # and inactive lever press number (e.g. VTA1G_ial1 -> Animal VTA1T inactive lever press 1)
Figure S6C: Maximum Th_cre GCaMP8f z-score associated with inactive lever press (analysis of traces in S6A)
same variables as S3C
- VTA#T_ial#: Animal ID # and inactive lever press number (e.g. VTA1G_ial1 -> Animal VTA1T inactive lever press 1)
Figure S7A: Lever responses during reinstatement for GCaMP8f injected Th-cre rats self-administering saline
VTA1TS-VTA3TS refers to animal ID ("VTA1_ThSaline")
Same as variables in 5C & S4A
Figure S7B: GCaMP trace during active lever press in Th_cre saline rats
Same as variables in 5D & S4B
Figure S7C: Net area under Th_cre GCaMP8f traces associated with active lever press (analysis of traces in S7B)
Same variables as 5E & S4C
Figure S7D: Maximum Gad_cre GCaMP8f z-score associated with active lever press (analysis of traces in S7B)
Same variables as 5F & S4D
Figure 8A: Th expression throughout VTA
- Animal ID: Rat subject ID name
- AP (mm): Anterior/posterior position of slice based off rat brain atlas according to Paxinos and Western (mm)
- # of th+ cells: # of th+ dapi+ cells
Figure 8b: GAD expression throughout VTA
- Animal ID: Rat subject ID name
- AP (mm): Anterior/posterior position of slice based off rat brain atlas according to Paxinos and Western (mm)
- # of Gad1+ cells: # of gad1+ dapi+ cells
Code/Software
Custom R Code used to analyze the fiberphotometry data in this study is accessible at: https://zenodo.org/records/13984378