Convergent, functionally independent signaling by mu and delta opioid receptors in hippocampal parvalbumin interneurons

  1. Xinyi Jenny He
  2. Janki Patel
  3. Connor E Weiss
  4. Xiang Ma
  5. Brenda L Bloodgood
  6. Matthew R Banghart  Is a corresponding author
  1. Division of Biological Sciences, Neurobiology Section, University of California, San Diego, United States
7 figures, 2 tables and 1 additional file

Figures

Figure 1 with 1 supplement
Electrophysiological recordings of opioid-sensitive synaptic output from hippocampal parvalbumin basket cells.

(A) Schematic of the experimental configuration for recording optogenetically evoked inhibitory synaptic transmission in PV-Cre mice. (B) Representative optically evoked IPSC (oIPSC) pairs (50 ms …

Figure 1—source data 1

IPSC suppression, paired pulse ratios, and time courses for DAMGO and SNC162.

https://cdn.elifesciences.org/articles/69746/elife-69746-fig1-data1-v3.xlsx
Figure 1—figure supplement 1
Opioid receptor mRNA in CA1 parvalbumin interneurons and characterization of the neuromodulator sensitivity of CA1 basket cell synaptic output.

(A) Example fluorescence in situ hybridization image of Pvalb, Oprm1, and Oprd1 mRNA in the CA1 pyramidal layer of mouse hippocampus. Scale bar = 20 μm. (B) Summary of Pvalb, Oprm1, and Oprd1 mRNA …

Figure 1—figure supplement 1—source data 1

IPSC suppression and time courses for WIN55,212, DAMGO, and SNC162 using opto and estim.

https://cdn.elifesciences.org/articles/69746/elife-69746-fig1-figsupp1-data1-v3.xlsx
Characterization of the potency and kinetics of synaptic modulation by [Leu5]-enkephalin (LE) at mu (MOR) and delta opioid receptors (DOR) using caged peptides.

(A) Left: Schematic of the experimental configuration for photo-uncaging of opioid neuropeptides while recording electrically evoked inhibitory synaptic transmission in wild-type mice. Right:

Figure 2—source data 1

Power-response curves and onset kinetics at presynaptic MOR and DOR.

https://cdn.elifesciences.org/articles/69746/elife-69746-fig2-data1-v3.xlsx
Axonal calcium imaging reveals that both mu and delta opioid receptors suppress presynaptic voltage-sensitive calcium channels.

(A) Two-photon image of a tdTomato-expressing basket cell filled with 30 µM Alexa 594 and 300 µM Fluo-5F in a brain slice taken from a PV-Cre; tdTom mouse. Scale bar: 50 μm. Inset shows the two …

Figure 3—source data 1

Ca2+ transient peaks with and without DAMGO and SNC162.

https://cdn.elifesciences.org/articles/69746/elife-69746-fig3-data1-v3.xlsx
Figure 4 with 1 supplement
Enkephalin evokes outward currents in CA1 parvalbumin (PV) interneurons through both mu and delta opioid receptors.

(A) Schematic of whole-cell voltage clamp recording configuration from PV interneurons with peptide uncaging. (B) Average outward currents evoked by photoactivation of N-MNVOC-LE (6 μM) with an 84 …

Figure 4—source data 1

Power-response curves and onset kinetics at somato-dendritic MOR and DOR and MOR currents after overexpression.

https://cdn.elifesciences.org/articles/69746/elife-69746-fig4-data1-v3.xlsx
Figure 4—figure supplement 1
Sensitivity of somato-dendritic currents to the G protein-coupled inward rectifier K+ (GIRK) blocker Ba2+ and mu opioid receptor expression level.

(A) Average outward currents evoked by photoactivation of CYLE with a 84 mW light flash in the absence (black, artificial cerebrospinal fluid [ACSF], n = 13 cells from seven mice) and presence of mu …

Figure 4—figure supplement 1—source data 1

Somato-dendritic currents in Ba2+ and ZD7288 and correlation between mCherry fluorescence and MOR currents.

https://cdn.elifesciences.org/articles/69746/elife-69746-fig4-figsupp1-data1-v3.xlsx
Somato-dendritic mu and delta opioid receptors do not exhibit heterologous desensitization.

(A) Average outward current evoked by sustained bath application of DAMGO (n = 9 cells from six mice). (B) Average outward currents evoked by photoactivation of N-MNVOC-LE either in the presence of …

Figure 5—source data 1

Somato-dendritic currents from DAMGO and Deltorphin II and power-response curves of uncaging-evoked currents.

https://cdn.elifesciences.org/articles/69746/elife-69746-fig5-data1-v3.xlsx
Figure 6 with 1 supplement
Mu and delta opioid receptors do not signal as heteromers in CA1 parvalbumin (PV) neurons.

(A) Schematic of whole-cell voltage clamp recording configuration from PV interneurons with peptide uncaging. (B) Average outward currents evoked by photoactivation of CNV-Y-DAMGO with an 84 mW …

Figure 6—source data 1

Power-response curves and onset kinetics of CNV-Y-DAMGO uncaging with and without TIPP-Psi.

https://cdn.elifesciences.org/articles/69746/elife-69746-fig6-data1-v3.xlsx
Figure 6—figure supplement 1
Optogenetic activation confirms that mu opioid receptor (MOR) and delta opioid receptor (DOR) do not signal as heteromers in parvalbumin (PV) terminals.

(A) Schematic of the experimental configuration for recording optogenetically evoked inhibitory synaptic transmission in PvalbCre mice. (B) Summary data showing fraction of optically evoked IPSC …

Figure 6—figure supplement 1—source data 1

Optogenetically-evoked IPSC suppression by 300 nM DAMGO with and without TIPP-Psi.

https://cdn.elifesciences.org/articles/69746/elife-69746-fig6-figsupp1-data1-v3.xlsx
Models of mu opioid receptor (MOR) and delta opioid receptor (DOR) signaling in the soma and the pre-synaptic terminal.

(A) In the soma, both MORs (blue) and DORs (red) signal through G protein-coupled inward rectifier K+ (GIRK) channels. MORs are expressed at lower levels than DORs, as the somato-dendritic currents …

Tables

Key resources table
Reagent type (species) or resourceDesignationSource or referenceIdentifiersAdditional information
Strain, strain background (Mus musculus, male and female)C57Bl/6The Jackson LaboratoryCat # 000664RRID:IMSR_JAX:000664
Strain, strain background (Mus musculus, male and female)PvalbCreThe Jackson LaboratoryCat # 012358RRID:IMSR_JAX:012358
Strain, strain background (Mus musculus, male and female)Rosa26-lsl-tdTomato (Ai14)The Jackson LaboratoryCat # 007914RRID:IMSR_JAX:007914
Recombinant DNA reagentAAV1-Syn-FLEX-Chronos-GFPAddgeneCat # 62722RRID:Addgene_62722
Recombinant DNA reagentAAVDJ-hSyn1-FLEX-mCh-T2A-FLAG-hMOR-WPREBanghart LabAddgene Plasmid #166970
Commercial assay or kitRNAscope Fluorescent Multiplex KitACD bio/Bio-TechneCat # 320850
Commercial assay or kitPvalb FISH probeACD bio/Bio-TechneCat # 421931-C3
Commercial assay or kitOprd1 FISH probeACD bio/Bio-TechneCat # 427371-C2
Commercial assay or kitOprm1 FISH probeACD bio/Bio-TechneCat # 315841
Chemical compound, drugN-MNVOC-LEBanghart et al., 2018
Chemical compound, drugCYLEBanghart Lab and NIDA Drug Supply Program Banghart and Sabatini, 2012MPSP-117 (NDSP)
Chemical compound, drugCNV-Y-DAMGOMa et al., 2021
Chemical compound, drugNBQXHelloBioCat # HB0443
Chemical compound, drug(R)-CPPHelloBioCat # HB0021
Chemical compound, drugTIPP-PsiNIDA Drug Supply ProgramMPSP-056
Chemical compound, drugCTOPTocrisCat # 1578
Chemical compound, drugDAMGOTocrisCat # 1171
Chemical compound, drugSNC162TocrisCat # 1529
Chemical compound, drugAlexaFluor 547Thermo FisherCat # 10438
Chemical compound, drugFluo5FThermo FisherCat # F14221
Chemical compound, drugPicrotoxinSigmaCat # P1675
Chemical compound, drugTTXHelloBioCat # HB1035
Chemical compound, drugWIN55,212TocrisCat # 1038
Chemical compound, drugDeltorphin IINIDA Drug Supply ProgramMPSP-036
Chemical compound, drugZD7288TocrisCat # 1000
Software, algorithmMATLABMathworks IncRRID:SCR_001622
Software, algorithmScanImagePologruto et al., 2003RRID:SCR_014307
Software, algorithmIgor ProWaveMetricsRRID:SCR_000325
Software, algorithmImageJNIHRRID:SCR_003070
Software, algorithmIllustrator CCAdobe Systems IncRRID:SCR_010279
Software, algorithmPrism 7GraphPad IncRRID:SCR_002798
Software, algorithmExcelMicrosoftRRID:SCR_016137
Author response table 1
FigureNormal?Results changed?
1DYesOne way ANOVANo
1E newNo for BL(DAM)Wilcoxon test, both significantNo
1IYesOne way ANOVANo
1J newNo for BL(DAM)Wilcoxon test, DAMGO p = 0.0186, SNC p = 0.058DAMGO significant, SNC no longer significant
S1EYesOne way ANOVA w/ Tukey (changed from Dunnett’s)No
S1GYesTwo way ANOVANo
S1HYesPaired t-testNew data
S1INoSkillings-Mack non-parametric test for grouped dataNew data
2GNoKruskal-Wallis (non-parametric): only significant difference is for 20hz, between ACSF and TIPP-PsiNo
3CNoFriedman test with Dunn’s multiple comparisonsNo
3DNoFriedman test with Dunn’s multiple comparisonsNo
4CNoKruskal-Wallis (non-parametric) with Dunn’s multiple comparisonsNo
4GNoKruskal-Wallis (non-parametric) with Dunn’s multiple comparisonsNo
4JYesUnpaired t-testNo
4KYesUnpaired t-testNo
S2BYesOne way ANOVA, and t-test for CTOP condition onlyNo
6CNoMann-Whitney test, p = 0.4252No
6HNoMann-Whitney test, p = 0.2824No
S3BYesUnpaired t-test, p = 0.7518No

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