Journal: The Journal of Clinical Investigation

Article Title: An endocannabinoid-regulated basolateral amygdala–nucleus accumbens circuit modulates sociability

doi: 10.1172/JCI131752

Figure Lengend Snippet: (A) Three-chamber SI task and optogenetic stimulation protocol. Animals were continuously stimulated during the 5-minute test using a 20-Hz pattern (10 mW, 5 seconds on and 5 seconds off). (B) Representative heatmaps of chamber time in a ChR2-expressing and YFP-expressing control mouse. (C) Optogenetic stimulation of the BLA-NAc circuit did not affect distance traveled (NS, P = 0.3224). (D) Animals that express ChR2 showed reduced social preference (**P = 0.0010, ****P < 0.0001, and NS, P = 0.5565), (E) decreased close interaction time (**P = 0.0011, ****P < 0.0001, and NS, P = 0.9353), and (F) reduced time investigating, or sniffing, the target mouse compared with animals that express YFP (M-M **P = 0.0026, E-M ****P < 0.001, and ##P = 0.0071). YFP n = 17, and ChR2 n = 13 (C–F). Data analyzed via 2-way mixed-effects ANOVA followed by Holm-Šídák multiple-comparisons test (D–F) or unpaired, 2-tailed t test (C). P and F values for chamber × virus interaction shown in D–F.

Article Snippet: In blue light stimulation studies, animals received 20-Hz blue light stimulation (Plexon) in a 5 seconds on, 5 seconds off, pattern at 10–13 mW.

Techniques: Expressing, Control, Virus

Journal: The Journal of Clinical Investigation

Article Title: An endocannabinoid-regulated basolateral amygdala–nucleus accumbens circuit modulates sociability

doi: 10.1172/JCI131752

Figure Lengend Snippet: (A) Optogenetic stimulation of the BLA-NAc circuit increased fleeing and withdrawal behaviors (***P = 0.0007) and (B) decreased social sniffing behaviors (**P = 0.0041) compared with YFP-expressing controls. There was no effect of BLA-NAc activation on (C) following (P = 0.1573) or (D) passive social behavior (P = 0.2604). Animals expressing ChR2 were significantly more (E) immobile (***P = 0.0007) and (F) explored less (*P = 0.0049) than YFP controls. (G) Animals that expressed ChR2 self-groomed significantly less than YFP-expressing controls (*P = 0.0184). YFP n = 15, and ChR2 n = 19 (A–G). (H) Social CPP paradigm. (I) Bilateral activation of the BLA-NAc circuit significantly decreased time in the social-paired chamber during the posttest (**P = 0.0013) (J) and reduced CPP score relative to YFP controls (**P = 0.0041). (K) Representative heatmaps for the social CPP experiment. (L) No pretest preference to either chamber was detected (YFP dots vs. stripes, P = 0.1777; ChR2 dots vs. stripes, P = 0.4619). YFP n = 9, and ChR2 n = 10 (I, J, L). Data analyzed via unpaired, 2-tailed t test (A–G, I, and J) or 2-way mixed-effects ANOVA with Holm-Šídák multiple-comparisons post hoc test (L), with P and F values for chamber × virus interaction shown in L.

Article Snippet: In blue light stimulation studies, animals received 20-Hz blue light stimulation (Plexon) in a 5 seconds on, 5 seconds off, pattern at 10–13 mW.

Techniques: Expressing, Activation Assay, Virus

Journal: The Journal of Clinical Investigation

Article Title: An endocannabinoid-regulated basolateral amygdala–nucleus accumbens circuit modulates sociability

doi: 10.1172/JCI131752

Figure Lengend Snippet: (A–C) Effects of BLA-NAc stimulation in the RtPP assay. (A) Representative heatmaps of RtPP results. (B and C) Animals expressing ChR2 spent significantly more time in the stimulation-paired (On) compared with nonpaired (Off) side in the RtPP assay (NS, P = 0.9083, and ****P < 0.0001) without any effect on total distance traveled (unpaired, 2-tailed t test, P = 0.0568). YFP n = 5, and ChR2 n = 9 (B and C). (D–G) Effects of BLA-NAc stimulation on Ensure-seeking behavior. (D) In a modified 3-chamber task a sipper bottle of Ensure was added to 1 chamber while stimulation was delivered to animals. (E) Activation of the BLA-NAc circuit did not alter time spent in the chamber with (***P = 0.0003, and ****P < 0.0001), (F) close to (*P = 0.0102, and **P = 0.0037), or (G) drinking (****P < 0.0001, and *P = 0.0128) the nutrition shake in ChR2-expressing animals compared to YFP-expressing animals. YFP n = 12, and ChR2 n = 9 (E–G). Data analyzed via unpaired, 2-tailed t test (C) or 2-way mixed-effects ANOVA with Holm-Šídák multiple-comparison post hoc tests (B, E–G), with P and F values for chamber × virus interaction shown in B, and E–G.

Article Snippet: In blue light stimulation studies, animals received 20-Hz blue light stimulation (Plexon) in a 5 seconds on, 5 seconds off, pattern at 10–13 mW.

Techniques: Expressing, Modification, Activation Assay, Comparison, Virus

Journal: The Journal of Clinical Investigation

Article Title: An endocannabinoid-regulated basolateral amygdala–nucleus accumbens circuit modulates sociability

doi: 10.1172/JCI131752

Figure Lengend Snippet: (A) The cannabinoid receptor agonist CP55940 decreased oEPSC amplitude in the NAc; N = 3. (B) JZL184 reduced aEPSC frequency in D1 receptor+ (D1R+; *P = 0.0171) and D1 receptor– (D1R–) cells (#P = 0.0197) (C) but did not affect aEPSC amplitude after BLA-NAc optogenetic stimulation; D1+ n = 3, and D1– n = 3 (B, C). aEPSC, asynchronous excitatory postsynaptic current; BL, baseline. (D) WIN55212-2 (Win55), a cannabinoid receptor agonist, uniformly decreased oEPSC amplitude in D1+ and D1– NAc cell types; D1+ n = 4, and D1– n = 4. (E) DSE was present in both D1+ and D1– NAc cells, (F) although DSE magnitude was increased in D1+ compared with D1– cells (*P = 0.0450); D1+ n = 4, and D1– n = 5 (E, F). (G) sEPSC frequency in NAc recordings were unaffected by JZL184, (H) but there was a significant effect of JZL184 on sIPSC frequency (*P = 0.0400). (I) There were no effects of JZL184 on sIPSC amplitude; D1+ n = 4, D1– n = 4. Data analyzed via 2-way mixed-effects ANOVA followed by Holm-Šídák multiple-comparisons test (B, C, G–I) or unpaired, 2-tailed t test (F). P and F values for drug effect shown in B, C, and G–I.

Article Snippet: In blue light stimulation studies, animals received 20-Hz blue light stimulation (Plexon) in a 5 seconds on, 5 seconds off, pattern at 10–13 mW.

Techniques:

Journal: The Journal of Clinical Investigation

Article Title: An endocannabinoid-regulated basolateral amygdala–nucleus accumbens circuit modulates sociability

doi: 10.1172/JCI131752

Figure Lengend Snippet: (A) Shank3B–/– mice expressing NpHR in the BLA with bilateral orange light stimulation delivered to the NAc spent significantly more time in the mouse chamber during orange light delivery (On) compared with light Off conditions (*P = 0.0230). YFP animals did not show a preference for the mouse chamber (NS, P = 0.2529). (B) Shank3B–/– mice expressing YFP under light On conditions did not exhibit social preference (NS, P = 0.2831), while animals that express NpHR had a preference for the mouse chamber (****P < 0.0001). (C) No effect on distance traveled was observed (NS, P = 0.9057) in the light On paradigm. (D) Mice that express NpHR had a significant increase in time investigating the mouse cup under light On conditions (****P < 0.0001). (E) Representative heatmaps under light On conditions. (F) Neither YFP (NS, P = 0.7166) nor NpHR (NS, P = 0.8731) Shank3B–/– mice showed mouse chamber preference (H) or preference for time investigating the mouse target (YFP NS, P = 0.1617; NpHR NS, P = 0.6193) under light Off conditions. (G) There were no effects on distance traveled under light Off conditions (P = 0.7959). (I) Representative heatmaps under light Off conditions. YFP n = 9, and NpHR n = 9 (B–D, F–H). Data analyzed via 2-way mixed-effects ANOVA with Holm-Šídák multiple-comparisons test (A, B, D, F, H) or unpaired, 2-tailed t test (C, G). P and F values for light × virus interaction shown in A and chamber × virus interaction shown in B, D, F, and H.

Article Snippet: In blue light stimulation studies, animals received 20-Hz blue light stimulation (Plexon) in a 5 seconds on, 5 seconds off, pattern at 10–13 mW.

Techniques: Expressing, Virus

Journal: The Journal of Clinical Investigation

Article Title: An endocannabinoid-regulated basolateral amygdala–nucleus accumbens circuit modulates sociability

doi: 10.1172/JCI131752

Figure Lengend Snippet: (A) sIPSC frequency on the NAc MSNs was significantly increased in Shank3B–/– compared with WT mice (**P = 0.0060) and was restored by JZL184 (##P = 0.0033). (B) There was no effect of Shank3B–/– on sIPSC amplitude. (C) sEPSC frequency was significantly increased in Shank3B–/– mice relative to WT animals (**P = 0.0084) and was restored by JZL184 (##P = 0.0060). (D) There was no effect of genotype or drug on sEPSC amplitude; WT + Veh n = 3, WT + JZL184 n = 3, KO + Veh n = 5, and KO + JZL184 n = 4 (A–D). (E) There was no difference between WT and Shank3B–/– BLA FF oIPSCs onto NAc cells. (F) However, at maximal stimulation (1.85 mW), JZL184 significantly reduced BLA FF oIPSCs onto NAc MSNs (*P = 0.0101). (G and H) There was no effect of JZL184 or genotype on BLA oEPSCs in the NAc across intensities; WT + Veh n = 3, WT + JZL184 n = 3, KO + Veh n = 4, and KO + JZL184 n = 4 (E–H). Data analyzed via 2-way ANOVA with Holm-Šídák multiple-comparisons test (A–H). P and F values for drug effect shown in A–H. M, males; F, females.

Article Snippet: In blue light stimulation studies, animals received 20-Hz blue light stimulation (Plexon) in a 5 seconds on, 5 seconds off, pattern at 10–13 mW.

Techniques: