(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 …
IPSC suppression, paired pulse ratios, and time courses for DAMGO and SNC162.
(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 …
IPSC suppression and time courses for WIN55,212, DAMGO, and SNC162 using opto and estim.
(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: …
Power-response curves and onset kinetics at presynaptic MOR and DOR.
(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 …
Ca2+ transient peaks with and without DAMGO and SNC162.
(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 …
Power-response curves and onset kinetics at somato-dendritic MOR and DOR and MOR currents after overexpression.
(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 …
Somato-dendritic currents in Ba2+ and ZD7288 and correlation between mCherry fluorescence and MOR currents.
(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 …
Somato-dendritic currents from DAMGO and Deltorphin II and power-response curves of uncaging-evoked currents.
(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 …
Power-response curves and onset kinetics of CNV-Y-DAMGO uncaging with and without TIPP-Psi.
(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 …
Optogenetically-evoked IPSC suppression by 300 nM DAMGO with and without TIPP-Psi.
(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 …
Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
---|---|---|---|---|
Strain, strain background (Mus musculus, male and female) | C57Bl/6 | The Jackson Laboratory | Cat # 000664RRID:IMSR_JAX:000664 | |
Strain, strain background (Mus musculus, male and female) | PvalbCre | The Jackson Laboratory | Cat # 012358RRID:IMSR_JAX:012358 | |
Strain, strain background (Mus musculus, male and female) | Rosa26-lsl-tdTomato (Ai14) | The Jackson Laboratory | Cat # 007914RRID:IMSR_JAX:007914 | |
Recombinant DNA reagent | AAV1-Syn-FLEX-Chronos-GFP | Addgene | Cat # 62722RRID:Addgene_62722 | |
Recombinant DNA reagent | AAVDJ-hSyn1-FLEX-mCh-T2A-FLAG-hMOR-WPRE | Banghart Lab | Addgene Plasmid #166970 | |
Commercial assay or kit | RNAscope Fluorescent Multiplex Kit | ACD bio/Bio-Techne | Cat # 320850 | |
Commercial assay or kit | Pvalb FISH probe | ACD bio/Bio-Techne | Cat # 421931-C3 | |
Commercial assay or kit | Oprd1 FISH probe | ACD bio/Bio-Techne | Cat # 427371-C2 | |
Commercial assay or kit | Oprm1 FISH probe | ACD bio/Bio-Techne | Cat # 315841 | |
Chemical compound, drug | N-MNVOC-LE | Banghart et al., 2018 | ||
Chemical compound, drug | CYLE | Banghart Lab and NIDA Drug Supply Program Banghart and Sabatini, 2012 | MPSP-117 (NDSP) | |
Chemical compound, drug | CNV-Y-DAMGO | Ma et al., 2021 | ||
Chemical compound, drug | NBQX | HelloBio | Cat # HB0443 | |
Chemical compound, drug | (R)-CPP | HelloBio | Cat # HB0021 | |
Chemical compound, drug | TIPP-Psi | NIDA Drug Supply Program | MPSP-056 | |
Chemical compound, drug | CTOP | Tocris | Cat # 1578 | |
Chemical compound, drug | DAMGO | Tocris | Cat # 1171 | |
Chemical compound, drug | SNC162 | Tocris | Cat # 1529 | |
Chemical compound, drug | AlexaFluor 547 | Thermo Fisher | Cat # 10438 | |
Chemical compound, drug | Fluo5F | Thermo Fisher | Cat # F14221 | |
Chemical compound, drug | Picrotoxin | Sigma | Cat # P1675 | |
Chemical compound, drug | TTX | HelloBio | Cat # HB1035 | |
Chemical compound, drug | WIN55,212 | Tocris | Cat # 1038 | |
Chemical compound, drug | Deltorphin II | NIDA Drug Supply Program | MPSP-036 | |
Chemical compound, drug | ZD7288 | Tocris | Cat # 1000 | |
Software, algorithm | MATLAB | Mathworks Inc | RRID:SCR_001622 | |
Software, algorithm | ScanImage | Pologruto et al., 2003 | RRID:SCR_014307 | |
Software, algorithm | Igor Pro | WaveMetrics | RRID:SCR_000325 | |
Software, algorithm | ImageJ | NIH | RRID:SCR_003070 | |
Software, algorithm | Illustrator CC | Adobe Systems Inc | RRID:SCR_010279 | |
Software, algorithm | Prism 7 | GraphPad Inc | RRID:SCR_002798 | |
Software, algorithm | Excel | Microsoft | RRID:SCR_016137 |
Figure | Normal? | Results changed? | |
---|---|---|---|
1D | Yes | One way ANOVA | No |
1E new | No for BL(DAM) | Wilcoxon test, both significant | No |
1I | Yes | One way ANOVA | No |
1J new | No for BL(DAM) | Wilcoxon test, DAMGO p = 0.0186, SNC p = 0.058 | DAMGO significant, SNC no longer significant |
S1E | Yes | One way ANOVA w/ Tukey (changed from Dunnett’s) | No |
S1G | Yes | Two way ANOVA | No |
S1H | Yes | Paired t-test | New data |
S1I | No | Skillings-Mack non-parametric test for grouped data | New data |
2G | No | Kruskal-Wallis (non-parametric): only significant difference is for 20hz, between ACSF and TIPP-Psi | No |
3C | No | Friedman test with Dunn’s multiple comparisons | No |
3D | No | Friedman test with Dunn’s multiple comparisons | No |
4C | No | Kruskal-Wallis (non-parametric) with Dunn’s multiple comparisons | No |
4G | No | Kruskal-Wallis (non-parametric) with Dunn’s multiple comparisons | No |
4J | Yes | Unpaired t-test | No |
4K | Yes | Unpaired t-test | No |
S2B | Yes | One way ANOVA, and t-test for CTOP condition only | No |
6C | No | Mann-Whitney test, p = 0.4252 | No |
6H | No | Mann-Whitney test, p = 0.2824 | No |
S3B | Yes | Unpaired t-test, p = 0.7518 | No |