Journal: Frontiers in Cellular Neuroscience

Article Title: The Basolateral Amygdala to Ventral Hippocampus Circuit Controls Anxiety-Like Behaviors Induced by Morphine Withdrawal

doi: 10.3389/fncel.2022.894886

Figure Lengend Snippet: BLA to vHip projections regulate anxiety-like behavior in morphine-withdrawn mice. (A–E) Chemogenetic experiment. (A) Workflow for the chemogenetic experiment. (B) Schematic (left) and representative image (right) of chemogenetic virus injection. Scale bar: 200 μm. (C) The Mor-A×CNO mice exhibited significantly increased central time and central distances in the OFT. (D) The Mor-A × CNO mice exhibited significantly increased open-arm times and entries in the EPM. (E) The hM4Di inhibition accelerated the decreased rate of morphine-paired preference in the Mor-A mice. (F–H) Optogenetic experiment. (F) Schematic of the virus injection site in the BLA and optical fiber implantation site in the vHip. (Right) Image of a coronal brain slice showing the expression of eNpHR-mCherry in the BLA. Scale bar: 200 μm. (G) Increased open-arm entries and time spent during the light-on epoch. (H) Increased time spent and distances traveled in the central area of the OFT during the eNpHR illumination epoch. The EPM (I) and OFT (J) tests of the saline + eNpHR group and the saline + mCherry group. Data are expressed as the mean ± SEM. In the chemogenetic experiment, morphine CNO-treated mice were compared with morphine DMSO-treated mice. In the CPP results, *: comparison between the extinction group and the post-test group, #: comparison between the 2 indicated groups. In the optogenetic results, #: comparison between the Mor+mcherry and the Mor+eNPHR group during the light on epoch. n = 8–10/group.* P < 0.05, ** P < 0.01, *** P < 0.001, # P < 0.05, ## P < 0.01, #### P < 0.0001.

Article Snippet: For optogenetic inhibition of BLA to vHip inputs, we used a constant illumination of yellow light stimulation (5 mw, NewDoon Aurora 220).

Techniques: Virus, Injection, Inhibition, Slice Preparation, Expressing, Saline, Comparison

Journal: Neurobiology of Stress

Article Title: Distinct populations of lateral preoptic nucleus neurons jointly contribute to depressive-like behaviors through divergent projections in male mice

doi: 10.1016/j.ynstr.2024.100667

Figure Lengend Snippet: Optogenetic activation of LHb-projecting LPO excitatory neurons produced aversion and anxiety-like behaviors in mice . (A) Experimental design to assess the effect of optogenetic activation of LHb-projecting LPO excitatory neurons on mice's behavioral performances. (B , C) Representative microphotographs showed the expression of Retro-hSyn-Cre-EGFP within LHb and AAV9-CaMKII-DIO-ChR2-mCherry in LPO. Scale bar: 100 μm and 250 μm, respectively. (D , E) Representative trajectories from the real-time place preference test. (F) Mice (n = 11) exhibited lower time in light-ON chamber (Two-way ANOVA with Sidak post-hoc test, test phase (Baseline, Real-time place test, Post) x chamber (light-ON vs. light-OFF) : F(2, 36) = 29.08, p < 0.0001; Test phase (Baseline, Real-time place test, Post) effect: F(1, 36) = 18.89, p = 0.0001; Chamber (light-ON vs. light-OFF) effect: F(2, 36) = 1.635, p = 0.2091; Ligh-OFF vs. light-ON chamber: Baseline, p = 0.9962; Real-time place test, p < 0.0001; Post: p = 0.6955). (G) Light stimulation had no effect on locomotor activity (Wilcoxon matched-pairs signed rank test, Baseline vs. Real-time place test: W = 27, p = 0.3757). (H) Representative trajectories from the elevated plus maze test. When optogenetically activating LHb-projecting LPO excitatory neurons (a 25-s off + 10-s on + 25-s off activation strategy), mice (n = 11) exhibited (I) decreased time (One-way ANOVA test, Treatment F(1.138, 11.38) = 12.58, p = 0.0035; OFF vs. ON: p = 0.01; ON vs. OFF, p = 0.0428), (J) decreased entry numbers in the open arms (One-way ANOVA test, Treatment F(1.604, 16.04) = 6.792, p = 0.0103; OFF vs. ON: p = 0.0317; ON vs. OFF, p = 0.4212), but no effect on (K) locomotor activity (One-way ANOVA test, Treatment F(1.778, 17.78) = 1.636, p = 0.2993; OFF vs. ON: p = 0.4521; ON vs. OFF, p = 0.2994). (L) Representative trajectories from the open field test. When the light was on, mice (n = 11) displayed (M) decreased time (One-way ANOVA test, Treatment F(1.64, 16.4) = 0.0339, p = 0.0339; OFF vs. ON: p = 0.0064; ON vs. OFF, p = 0.2238) and (N) decreased center entries in the center area (One-way ANOVA test, Treatment F(1.682, 16.82) = 22.69, p < 0.001; OFF vs. ON: p < 0.0001; ON vs. OFF, p = 0.012), but no effect on (O) locomotor activity (One-way ANOVA test, Treatment F(1.99, 19.9) = 0.01263, p = 0.9872; ON vs. OFF, p = 0.9973). *p < 0.05, ***p < 0.001, N.S. not significant. Values are expressed as mean ± sem.

Article Snippet: For elevated plus maze and open field tests, mice were recorded with first 3 min as the baseline, followed by second 3 min’ delivery of 473-nm light stimulation with 10 ms, 10 Hz (Newdoon Technology) via an optic cable (200-μm core, 0.37 NA, Doric Lens).

Techniques: Activation Assay, Produced, Expressing, Activity Assay

Journal: Neurobiology of Stress

Article Title: Distinct populations of lateral preoptic nucleus neurons jointly contribute to depressive-like behaviors through divergent projections in male mice

doi: 10.1016/j.ynstr.2024.100667

Figure Lengend Snippet: Optogenetic activation of VTA-projecting LPO excitatory neurons produced preference and risk-taking behaviors in mice . (A) Experimental design to assess the effect of optogenetic activation of VTA-projecting LPO excitatory neurons on mice's behavioral performances. (B , C) Representative microphotographs showed the expression of Retro-hSyn-Cre-EGFP within VTA and AAV9-CaMKII-DIO-ChR2-mCherry in LPO. Scale bar: 250 μm. (D , E) Representative trajectories from the real-time place preference test. (F) Mice (n = 12) exhibited more time in light-ON chamber (Two-way ANOVA with Sidak post-hoc test, test phase (Baseline, Real-time place test, Post) x chamber (light-ON vs. light-OFF) : F(2, 33) = 36.76, p < 0.0001; test phase (Baseline, Real-time place test, Post) effect: F(1, 33) = 22.72, p < 0.0001; chamber (light-ON vs. light-OFF) effect: F(2, 23) = 0.0004, p = 0.9996; Ligh-OFF vs. light-ON chamber: Baseline, p = 0.7191; Real-time place test, p < 0.0001; Post: p = 0.9343), and (G) light stimulation had no effect on locomotor activity (Wilcoxon matched-pairs signed rank test, Baseline vs. Real-time place test: W = 30, p = 0.2661). (H) Representative trajectories from the elevated plus maze test. When optogenetically activating VTA-projecting LPO excitatory neurons (a 25-s off + 10-s on + 25-s off activation strategy), mice (n = 12) exhibited (I) increased time (nonparametric followed by Dunnett's test, p = 0.0024; OFF vs. ON: p = 0.0416; ON vs. OFF, p = 0.0024), (J) increased entry numbers in the open arms (One-way ANOVA test, Treatment F(1.441, 15.85) = 30.32, p < 0.001; OFF vs. ON: p = 0.0004; ON vs. OFF, p = 0.0002), but no effect on (K) locomotor activity (One-way ANOVA test, Treatment F(1.221, 13.43) = 0.9246, p = 0.3735; OFF vs. ON: p = 0.4205; ON vs. OFF, p = 0.3516). (L) Representative trajectories from the open field test. When the light was on, mice (n = 12) displayed (M) increased time (nonparametric followed by Dunnett's test, p = 0.0204; OFF vs. ON: p = 0.0206; ON vs. OFF, p = 0.1542), (N) increased center entries in the center area (nonparametric followed by Dunnett's test, p = 0.0001; OFF vs. ON: p = 0.0008; ON vs. OFF, p = 0.0005), but no effect on (O) locomotor activity (nonparametric followed by Dunnett's test, p = 0.9733; OFF vs. ON: p > 0.9999; ON vs. OFF, p > 0.9999). *p < 0.05, **p < 0.01, ***p < 0.001, N.S. not significant. Values are expressed as mean ± sem.

Article Snippet: For elevated plus maze and open field tests, mice were recorded with first 3 min as the baseline, followed by second 3 min’ delivery of 473-nm light stimulation with 10 ms, 10 Hz (Newdoon Technology) via an optic cable (200-μm core, 0.37 NA, Doric Lens).

Techniques: Activation Assay, Produced, Expressing, Activity Assay

Journal: Neurobiology of Stress

Article Title: Distinct populations of lateral preoptic nucleus neurons jointly contribute to depressive-like behaviors through divergent projections in male mice

doi: 10.1016/j.ynstr.2024.100667

Figure Lengend Snippet: Optogenetic inhibition of LHb-projecting or activation of VTA-projecting LPO excitatory neurons during chronic restraint stress prevented depressive-like behaviors in mice . (A) Experimental design to assess the effect of optogenetic inhibition of LHb-projecting LPO excitatory neurons during chronic restraint stress on depressive-like behaviors in the CRS-induced depression mouse model. (B , C) Representative virus expression in the LHb and LPO. Using a 3-min off + 3-min on + 3-min off optogenetic stimulation paradigm, mice (n = 15 for control group, n = 13 for CRS group, and n = 8 for CRS + inhibition group) throughout optogenetic inhibition during chronic restraint stress displayed (D) increased time spent in open arms (One-way ANOVA test with Tukey post-hoc test, Treatment F(2, 23) = 11.22, p = 0.0002; Ctrl vs. CRS: p = 0.0011; CRS vs. CRS + inhibition, p = 0.0007) and (E) increased number of entries into open arms in the elevated plus maze test (One-way ANOVA test with Tukey post-hoc test, Treatment F(2, 33) = 6.542, p = 0.004; Ctrl vs. CRS: p < 0.0001; CRS vs. CRS + inhibition, p = 0.0056), (F) increased center time (One-way ANOVA test with Tukey post-hoc test, Treatment F(2, 33) = 14.49, p < 0.001; Ctrl vs. CRS: p < 0.001; CRS vs. CRS + inhibition, p = 0.0056) but no effect on (G) number of entries in the center in the open field test (One-way ANOVA test with Tukey post-hoc test, Treatment F(2, 33) = 17.98, p < 0.0001; Ctrl vs. CRS: p < 0.0001; CRS vs. CRS + inhibition, p = 0.466). And (H) increased percentage of sucrose intake in sucrose preference test (One-way ANOVA test with Tukey post-hoc test, Treatment F(2, 33) = 5.825, p = 0.0068; Ctrl vs. CRS: p = 0.0297; CRS vs. CRS + inhibition, p = 0.0109) while (I) decreased immobile time in forced swim test (One-way ANOVA test with Tukey post-hoc test, Treatment F(2, 33) = 4.406, p = 0.0201; Ctrl vs. CRS: p = 0.0459; CRS vs. CRS + inhibition, p = 0.039). (J) Experimental design to assess the effect of optogenetic activation of VTA-projecting LPO excitatory neurons during chronic restraint stress on depressive-like behaviors in the CRS-induced depression mouse model. (K , L) Representative virus expression in VTA and LPO. Mice (n = 15 for control group, n = 13 for CRS group, and n = 9 for CRS + activation group) after optogenetic activation during chronic restraint stress displayed (M) increased time spent in open arms (One-way ANOVA test with Tukey post-hoc test, Treatment F(2, 34) = 7.561, p = 0.0019; Ctrl vs. CRS: p = 0.0242; CRS vs. CRS + activation, p = 0.0021) and (N) increased number of entries into open arms in the elevated plus maze test (One-way ANOVA test with Tukey post-hoc test, Treatment F(2, 34) = 12.07, p = 0.0001; Ctrl vs. CRS: p = 0.5592; CRS vs. CRS + activation, p = 0.0001), (O) no effect on the time in the center (One-way ANOVA test with Tukey post-hoc test, Treatment F(2, 34) = 3.343, p = 0.0473; Ctrl vs. CRS: p = 0.0385; CRS vs. CRS + activation, p = 0.313) but (P) increased number of entries in the center in the open field test (One-way ANOVA test with Tukey post-hoc test, Treatment F(2, 34) = 4.52, p = 0.0182; Ctrl vs. CRS: p = 0.0453; CRS vs. CRS + activation, p = 0.0317). And (Q) increased percentage of sucrose intake in sucrose preference test (One-way ANOVA test with Tukey post-hoc test, Treatment F(2, 34) = 7.217, p = 0.0024; Ctrl vs. CRS: p = 0.0023; CRS vs. CRS + activation, p = 0.0363) while (R) decreased immobile time in forced swim test (One-way ANOVA test with Tukey post-hoc test, Treatment F(2, 34) = 14.01, p < 0.0001; Ctrl vs. CRS: p < 0.0001; CRS vs. CRS + activation, p = 0.0112). *p < 0.05, **p < 0.01, ***p < 0.001, N.S. not significant. Values are expressed as mean ± sem.

Article Snippet: For elevated plus maze and open field tests, mice were recorded with first 3 min as the baseline, followed by second 3 min’ delivery of 473-nm light stimulation with 10 ms, 10 Hz (Newdoon Technology) via an optic cable (200-μm core, 0.37 NA, Doric Lens).

Techniques: Inhibition, Activation Assay, Virus, Expressing, Control