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

Journal: Nature Communications

Article Title: Optimized design and in vivo application of optogenetically functionalized Drosophila dopamine receptors

doi: 10.1038/s41467-023-43970-0

Figure Lengend Snippet: a OptoPAD setup allowing light stimulation upon feeding action. Flies expressing optoXRs in a subset of MBONs (MBONγ5β’2a,β2mp and β2mp-bilateral) related to behavioral valence receive a light stimulus (1 s 525 nm 400 μW/cm 2 ) every time they feed on the sucrose drop. b Cumulative sips over time for flies expressing optoDop1R2 V2 using MB011B-Gal4 without or with light stimulation (mean ± SEM, n = 58, 63 animals). c Total sips at 60 min for flies expressing optoDop1R2 V2 using MB011B-Gal4 without or with light stimulation ( n = 58, 63 animals, two-tailed Mann-Whitney test). d Total sips at 60 min for flies expressing optoDop1R1 V2 using MB011B-Gal4 without or with light stimulation ( n = 65, 65 animals, two-tailed Mann-Whitney test). e Total sips at 60 min for flies expressing Dop1R2 RNAi control or with MB011B-Gal4 ( n = 54, 50 animals, two-tailed Mann-Whitney test). f Total sips at 60 min for flies expressing Dop1R1 RNAi control or with MB011B-Gal4 ( n = 41, 47 animals, two-tailed Mann-Whitney test). All boxplots depict 75th (top), median (central line) and 25th (bottom) percentile, whiskers depict 99th (top) and 1st (bottom) percentile. All violin plots with single data points depict data distribution, dotted lines depict 75th (top) and 25th (bottom) percentile, solid central line the median. Source data and statistical details are provided as a Source Data file.

Article Snippet: For activation of the optoDop1R2 V2 , larvae were subjected to 10s blue light stimulation (470 nm, 720 μW/cm2, CoolLED) twice with an interval of 30s between each pulse.

Techniques: Expressing, Two Tailed Test, MANN-WHITNEY, Control

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