(A) Schematic of optogenetic manipulation. (B) Timeline of experiments. (C) Immunohistological image showing virus expression in the DR (c1) and amplified images in the left box showing the mCherry, …
Source data indicating behavioral performances of bidirectional optogenetic modulation of 5-HT neurons in the DR in the DR-ACC neural circuit.
The right panels show colocalization of TPH2+ neurons (green) and CTB (red) in the DR. Scale bars, 100 μm. M: male; F: female; ACC: anterior cingulate cortex; DR: dorsal raphe nucleus; TPH2: …
(a1 and b1): Merged image of DAPI and mCherry; (a2–a4, b2–b4): amplified images in the left box showing the mCherry, TPH2, and the colocalization of mCherry and TPH2; (a5, b5): quantification rates …
(A) Quantification of chasing time in the consolation test of the CHR2 and control animals (n = 8 in each group; independent samples t-test and Bayesian independent samples t-test, two tailed; male: …
Source data indicating bidirectional optogenetic modulation of DR 5-HT neuron activities in the DR-ACC neural circuit on some control behaviors.
(A) Time spent in allogrooming (male: t(7) = −11.707, p < 0.001, BF+0 = 142.924; female: t(7) = −7.427, p<0.001, BF+0 = 400.207). (B) Allogrooming frequency (male: t(7) = −8.104, p<0.001, BF+0 = …
Source data indicating the optogenetic inhibition of DR 5-HT neurons in the DR-ACC neural circuit does not elicit long-lasting effects.
(A) Schematic of optogenetic manipulation. (B) Timeline of experiments. (C) Quantification of time spent in allogrooming in the consolation test of the CHR2 and control animals (n = 8 in each group; …
Source data indicating behavioral performances of optogenetic bidirectional modulation of 5-HT terminals within the ACC in the DR-ACC neural circuit.
(A) Quantification of chasing time in the consolation test of the CHR2 and control animals (n = 8 in each group; independent samples t-test and Bayesian independent samples t-test, two-tailed; male: …
Source data indicating optogenetic bidirectional modulation of ACC 5-HT terminals in the DR-ACC neural circuit on some control behaviors.
(A) Time spent in allogrooming (male: t(6) = −19.792, p < 0.001, BF+0 = 21609.467; female: t(7) = −10.404, p < 0.001, BF+0 = 2475.291). (B) Allogrooming frequency (male: t(6) = −11.210, p < 0.001, …
Source data indicating the optogenetic inhibition of ACC 5-HT terminals in the DR-ACC neural circuit does not elicit long-lasting effects.
(A) Schematic of chemogenetical manipulations. (B) Timeline of experiments. (C) Immunohistological image showing virus expression in the DR (c1) and amplified images in the left white box showing …
Source data indicating chemogenetic modulation of DR 5-HT neuron activities in the DR-ACC neural circuit.
(a1 and b1) Merged image of DAPI and mCherry; (a2–a4, b2–b4) amplified images in the left box showing the mCherry, TPH2, and the colocalization of mCherry and TPH2; (a5, b5) quantification rates of …
(A) Quantification of chasing time in the consolation test (male: treatment: F(1, 19) = 2.118, p = 0.162, BF(incl) = 0.757; group: F(2, 19) = 11.238, p<0.001, BF(incl) = 7.968; treatment × group: F(1…
Source data indicating the effects of chemogenetic modulation of DR 5-HT neuron activities in the DR-ACC neural circuit on some control behaviors.
(A) Schematic diagrams depicting the virus injection and recording sites. (B) Histology showing the expression of GCaMP6 (left) and GFP control (right) in the DR. (C) Experimental timeline for …
Source data indicating GCaMP6 and GFP fluorescent signals align to some behaviors.
(a1) Immunohistological image showing GCaMP6 expression in the DR; (a2–a4) amplified images in the left box showing the GCaMP6 (green), TPH2 (red), and the colocalization (yellow) of the two; (a5) …
(A1, B1, C1) Representative peri-event plot of GCaMP6 fluorescence signals aligned to the end of allogrooming, social approaching, and sniffing (for all peri-event plots, the red line denotes the …
Source data indicating GCaMP6 fluorescent signals align to the end of some behaviors.
(A) Schematic diagrams depicting the virus injection and recording sites. (B) Histology showing the expression of 5-HT sensor (left) and GFP control (right) within the ACC. (C) Experimental timeline …
Source data indicating 5-HT sensor and GFP fluorescent signals align to some behaviors.
(A1, B1, C1) Representative peri-event plot of 5-HT sensor fluorescence signals aligned to the end of allogrooming, social approaching, and sniffing (for all peri-event plots, the blue line denotes …
Source data indicating 5-HT sensor fluorescent signals align to the end of some behaviors.
(A) Schematic diagrams. (B) Combined individual 5-HT sensor fluorescence traces during allogrooming in response to Met (blue) or Veh (red). (C) Quantification of 5-HT sensor fluorescence signals …
Source data indicating 5-HT sensor fluoresce changes during allogrooming could be blocked by Met.
(A) Schematic representation of ACC infusion sites and virus strategy. (B) Timeline of experimental design. (C) Effect of a 5-HT1AR agonist 8-OH-DPAT on allogrooming time in the consolation test (n =…
Source data indicating 8-OH-DPAT but not MDL 100907 reduced sociability deficits induced by chemogenetic inhibition of DR 5-HT neurons in the DR→ACC neural circuit.
(A) Male: N = 5 in each group. (Vehicle + Saline)-(Vehicle + CNO) vs (8-OH-DPAT+Saline)-(8-OH-DPAT+CNO): t(8) = 2.375, p = 0.045, BF+0 = 3.750 with median posterior δ = 0.999, 95% CI = [0.091 to …
Source data indicating pretreatment with 8-OH-DPAT but not MDL 100907 significantly reduced the impact of CNO.
Injection of 5-HT1AR agonist (8-OH-DPAT) or 5-HT2AR antagonist (MDL 100907) had no significant effect on control behaviors of selfgrooming (A, C) and distance traveled in the open-field test (B, D). …
Source data indicating the treatments had no significant effect on control behaviors of selfgrooming and the distance traveled in the open-field test.
Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
---|---|---|---|---|
Antibody | Anti-TPH2 (goat polyclonal antibody) | Abcam | ab121013; RRID:AB_10898794 | IF (1/500) |
Antibody | Alexa Fluor 488-AffiniPure Donkey Anti-Goat IgG (H + L) | Jackson | 705-545-147 RRID:AB_2336933 | IF (1/300) |
Antibody | Rhodamine (TRITC)-AffiniPure Donkey Anti-Goat IgG (H + L) | Jackson | 705-025-003 RRID:AB_2340388 | IF (1/300) |
Chemical compound, drug | 8-OH-DPAT | Sigma | H8520 | Injection site: ACC; volume: 200 nl/side; concentration:1.5 mg/ml |
Chemical compound, drug | MDL 100907 | Tocris Bioscience | Cat# 4173 | Injection site: ACC; volume:200 nl/side; concentration:1 mg/ml |
Chemical compound, drug | Metergoline | MedChemExpress | Cat# HY-B1033/CS-4551 | 4 mg/kg, i.p. |
Chemical compound, drug | CNO | BrainVTA | Cat# CNO-02 | 1 mg/kg, i.p. |
Chemical compound, drug | Cholera Toxin Subunit B (CTB)594 | Thermo Fisher | Cat# C34777 | Injection site: ACC; volume:400 nl/side; expression: 10 days |
Transfected construct (human) | rAAV(Retro)-TPH2-Cre-WPRE-pA | BrainVTA | R-396-K181127 | Serotype: AAV2; titer: 5.86 × 1012 vg/ml; injection site: ACC; volume: 300 nl/side |
Transfected construct (human) | rAAV-Ef1α-DIO-ChR2-mCherry-WPRE-pA | BrainVTA | 9–2 K190827 | Serotype: AAV2/9; titer: 2.01 × 1012 vg/ml; injection site: DR; volume: 500 nl |
Transfected construct (human) | rAAV-Ef1α-DIO-eNpHR3.0-mCherry-WPRE-pA | BrainVTA | rAAV9-7-5-1 | Serotype: AAV2/9; titer: 2.23 × 1012 vg/ml; injection site: DR; volume: 500 nl |
Transfected construct (human) | rAAV-Ef1α-DIO-mCherry-WPRE-pA | BrainVTA | 9–13 K190430 | Serotype: AAV2/9; titer: 2.02 × 1012 vg/ml; injection site: DR; volume: 500 nl |
Transfected construct (human) | rAAV-Ef1α-DIO-hM4Di(Gi)-mCherry-WPRE-pA | BrainVTA | 9–43 K190521 | Serotype: AAV2/9; titer: 2.09 × 1012 vg/ml; injection site: DR; volume: 500 nl |
Transfected construct (human) | rAAV-Ef1α-DIO-hM4Dq(Gq)-mCherry-WPRE-pA | BrainVTA | 9–42 K190530 | Serotype: AAV2/9; titer: 2.70 × 1012 vg/ml; injection site: DR; volume: 500 nl |
Transfected construct (human) | rAAV-hSyn-5HT2.1-WPRE-hGHpA (5-HT sensor) | BrainVTA | 9–1826 K190620 | Serotype: AAV2/9; titer: 2.53 × 1012 vg/ml; injection site: ACC; volume: 400 nl |
Transfected construct (human) | rAAV-hSyn-EGFP-WPRE-hGH-pA | BrainVTA | PT-1990 | Serotype: AAV2/9; titer: 2.00 × 1012 vg/ml; injection site: ACC; volume: 400 nl |
Transfected construct (human) | rAAV-EF1α-DIO-GCaMp6-WPRE-hGH pA | BrainVTA | PT-0071 | Serotype: AAV2/9; titer: 5.15 × 1012 vg/ml; injection site: DR; volume: 500 nl |
Transfected construct (human) | rAAV-hSyn-DIO-EGFP-WPRE-hGH-PA | BrainVTA | 9–1103 K200409 | Serotype: AAV2/9; titer: 2.00 × 1012 vg/ml; injection site: DR; volume: 500 nl |
Software, algorithm | JASP | JASP | RRID:SCR_015823 | |
Other | DAPI stain | Boster | AR1177 | 1 µg/ml |