Journal: Neural Regeneration Research

Article Title: How do lateral septum projections to the ventral CA1 influence sociability?

doi: 10.4103/1673-5374.389304

Figure Lengend Snippet: Acute or chronic social interaction activates LS GABA neurons. (A) Diagram showing the acute social interaction task in the home cage and the experimental timeline. Illustrations were created with BioRender.com. (B) Representative images and quantification of c-Fos-positive (green, Alexa Fluor® 488) cell number in the LS of mice without or with a target mouse during the acute social interaction task ( n = 20 slices from five mice/group). The c-Fos-positive cell number was increased significantly in the target group compared with that in the without target group (two-tailed unpaired t -test). (C) Double immunofluorescence images indicating the co-localization of c-Fos-positive cells (green, Alexa Fluor® 488) and GABAergic neurons (red, Alexa Fluor® 546), as well as the percentage of c-Fos-positive/GABAergic co-localized neurons in the total group of c-Fos-positive cells in the LS of mice without or with a target mouse during the acute social interaction task ( n = 24 slices from six mice/group). We found no significant difference between the two groups (two-tailed unpaired t -test). Scale bars: 100 μm at low magnification and 25 μm at high magnification. (D) Representative traces of AP activity, AP frequency (Mann-Whitney U test), AP amplitude (Mann-Whitney U test), and membrane potential (two-tailed unpaired t -test with Welch's correction) of GABAergic neurons in the LS of mice without or with a target during the acute social interaction task (without target group: n = 18 neurons from three mice; with target group: n = 17 neurons from three mice). (E) Diagram showing the chronic social interaction task in the home cage and the experimental timeline. (F) Representative traces of AP activity, AP frequency (two-tailed unpaired t -test with Welch's correction), AP amplitude (two-tailed unpaired t -test), and membrane potential (two-tailed unpaired t -test) of GABAergic neurons in the LS of mice without or with a target during the chronic social interaction task (without target group: n = 17 neurons from three mice; with target group: n = 16 neurons from three mice). Data are expressed as mean ± SEM. * P < 0.05, ** P < 0.01, *** P < 0.001, vs. AAV-mCherry group. AP: Action potential; Fos: FBJ murine osteosarcoma viral oncogene homolog; GABA: gamma-aminobutyric acid; LS: lateral septum; LV: lateral ventricle.

Article Snippet: All sections were kept at 4°C overnight in a blocking solution containing the following primary antibodies: rabbit anti-c-FBJ murine osteosarcoma viral oncogene homolog (Fos; 1:1000, CST, Danvers, MA, USA, Cat# 2250S, RRID: AB_2247211), rabbit anti-ΔFosB (1:1000, CST, Cat# 2251S, RRID: AB_2106903), rabbit anti-GABA (1:500, Sigma-Aldrich, Cat# A2052, RRID: AB_477652), and mouse anti-vesicular glutamate transporter 2 (Vglut2; 1:400, Abcam, Cambridge, UK, Cat# ab79157, RRID: AB_1603114).

Techniques: Two Tailed Test, Immunofluorescence, Activity Assay, MANN-WHITNEY, Membrane

Journal: Neural Regeneration Research

Article Title: How do lateral septum projections to the ventral CA1 influence sociability?

doi: 10.4103/1673-5374.389304

Figure Lengend Snippet: Inhibition of LS GABA neurons suppressed social behaviors. (A) Representative image showing the expression of AAV-DIO-hM4-mCherry in the LS of a Vgat-Cre mouse and the experimental timeline. (B) Upper panel: schematic diagram showing the setup of the social preference task and the representative activity tracking data from the AAV-mCherry and AAV-hM4 mice in the three-chamber test. Lower panel: the time spent in the chambers (two-way repeated-measures analysis of variance with Sidak's test), social interaction index (two-tailed unpaired t -test), duration of sniffing (two-way repeated-measures analysis of variance with Sidak's test), and the discrimination index (two-tailed unpaired t -test with Welch's correction) of AAV-mCherry and AAV-hM4 mice in the three-chamber test. (C) Upper panel: Schematic diagram showing the setting of the social novelty task and the representative activity tracking data from the three-chamber test. Lower panel: The time spent in the chambers (two-way repeated-measures analysis of variance with Sidak's test), social interaction index (two-tailed unpaired t-test with Welch's correction), duration of sniffing (two-way repeated-measures analysis of variance with Sidak's test), and the discrimination index (Mann Whitney U test) in the three-chamber test (AAV-mCherry group, n = 7; AAV-hM4 group, n = 8). (D) Left panel: representative images showing c-Fos-positive cells in the LS in AAV-mCherry and AAV-hM4 mice subjected to 30 minutes of restraint. Right panel: Quantitative analysis of the number of c-Fos-positive cells in the LS (two-tailed unpaired t -test; AAV-mCherry group, n = 8 slices from three mice; AAV-hM4 group, n = 10 slices from three mice). (E) Upper panel: Representative traces of AP activity from LS GABAergic neurons; lower panel: AP frequency (two-tailed unpaired t -test with Welch's correction) and AP amplitude (two-tailed unpaired t -test) in AAV-mcherry and AAV-hM4 mice (AAV-mCherry group, n = 11 slices from three mice; AAV-hM4 group, n = 10 slices from three mice). Scale bars: 100 μm at low magnification and 25 μm at high magnification. Data are expressed as mean ± SEM. * P < 0.05, ** P < 0.01, *** P < 0.001, vs. AAV-mCherry group. AAV: Adeno-associated viruses; AP: action potential; CNO: clozapine-N-oxide; DIO: double-floxed inverted open reading frame; Fos: FBJ murine osteosarcoma viral oncogene homolog; GABA: gamma-aminobutyric acid; hM4: hM4D(Gi); LS: lateral septum; LV: lateral ventricle; Vgat: vesicular GABA transporter.

Article Snippet: All sections were kept at 4°C overnight in a blocking solution containing the following primary antibodies: rabbit anti-c-FBJ murine osteosarcoma viral oncogene homolog (Fos; 1:1000, CST, Danvers, MA, USA, Cat# 2250S, RRID: AB_2247211), rabbit anti-ΔFosB (1:1000, CST, Cat# 2251S, RRID: AB_2106903), rabbit anti-GABA (1:500, Sigma-Aldrich, Cat# A2052, RRID: AB_477652), and mouse anti-vesicular glutamate transporter 2 (Vglut2; 1:400, Abcam, Cambridge, UK, Cat# ab79157, RRID: AB_1603114).

Techniques: Inhibition, Expressing, Activity Assay, Two Tailed Test, MANN-WHITNEY

Journal: Neural Regeneration Research

Article Title: How do lateral septum projections to the ventral CA1 influence sociability?

doi: 10.4103/1673-5374.389304

Figure Lengend Snippet: The activation of LS GABA neurons increases social behaviors. (A) Representative image showing the expression of AAV-DIO-hM3-mCherry in the LS of a Vgat-Cre mouse and the experimental timeline. (B) Upper panel: Schematic showing the social preference task and the representative activity tracking data of AAV-mCherry and AAV-hM3 mice in the three-chamber test. Lower panel: The time spent in the chambers (two-way repeated-measures analysis of variance with Sidak's test), the social interaction index (two-tailed unpaired t -test), duration of sniffing (two-way repeated-measures analysis of variance with Sidak's test), and the discrimination index (Mann-Whitney U test) of AAV-mCherry and AAV-hM3 mice in the three-chamber test. (C) Upper panel: Schematic diagram showing the social novelty task and the representative activity tracking data in the three-chamber test. Lower panel: The time spent in the chambers (two-way repeated-measures analysis of variance with Sidak's test), the social interaction index (two-tailed unpaired t -test), duration of sniffing (two-way repeated-measures analysis of variance with Sidak's test), and the discrimination index (two-tailed unpaired t -test) in the three-chamber test (AAV-mCherry group, n = 7; AAV-hM3 group, n = 9). (D) Left panel: Representative images showing c-Fos-positive cells in the LS of AAV-mCherry and AAV-hM3 mice. Right panel: Quantitative analysis of the number of c-Fos-positive cells in the LS (two-tailed unpaired t -test with Welch's correction; AAV-mCherry group, n = 7 slices from three mice; AAV-hM3 group, n = 9 slices from three mice). Upper panel: Representative traces of AP activity from LS GABAergic neurons; lower panel: AP frequency (two-tailed unpaired t -test) and AP amplitude (two-tailed unpaired t -test with Welch's correction) in AAV-mcherry and AAV-hM3 mice (AAV-mCherry group, n = 11 slices from three mice; AAV-hM3 group, n = 11 slices from three mice). Scale bars: 100 μm at low magnification and 25 μm at high magnification. Data are expressed as mean ± SEM. * P < 0.05, ** P < 0.01, *** P < 0.001, vs . AAV-mCherry group. AAV: Adeno-associated viruses; AP: action potential; CNO: clozapine-N-oxide; DIO: double-floxed inverted open reading frame; Fos: FBJ murine osteosarcoma viral oncogene homolog; GABA: gamma-aminobutyric acid; hM3: hM3D(Gq); LS: lateral septum; LV: lateral ventricle; Vgat: vesicular GABA transporter.

Article Snippet: All sections were kept at 4°C overnight in a blocking solution containing the following primary antibodies: rabbit anti-c-FBJ murine osteosarcoma viral oncogene homolog (Fos; 1:1000, CST, Danvers, MA, USA, Cat# 2250S, RRID: AB_2247211), rabbit anti-ΔFosB (1:1000, CST, Cat# 2251S, RRID: AB_2106903), rabbit anti-GABA (1:500, Sigma-Aldrich, Cat# A2052, RRID: AB_477652), and mouse anti-vesicular glutamate transporter 2 (Vglut2; 1:400, Abcam, Cambridge, UK, Cat# ab79157, RRID: AB_1603114).

Techniques: Activation Assay, Expressing, Activity Assay, Two Tailed Test, MANN-WHITNEY

Journal: Neural Regeneration Research

Article Title: How do lateral septum projections to the ventral CA1 influence sociability?

doi: 10.4103/1673-5374.389304

Figure Lengend Snippet: Re-inhibition of socially activated LS GABA neurons attenuates social behaviors. (A) Representative image showing the expression of AAV-DIO-hM4-mCherry in the LS and the experimental timeline. Illustrations were created with BioRender.com. (B) Upper panel: Schematic diagram showing the social preference task and the representative tracking data of AAV-mCherry and AAV-hM4 mice in the three-chamber test. Lower panel: The time spent in the chambers (two-way repeated-measures analysis of variance with Sidak's test), the social interaction index (two-tailed unpaired t -test), sniffing time (two-way repeated-measures analysis of variance with Sidak's test), and the discrimination index (two-tailed unpaired t -test) of AAV-mCherry and AAV-hM4 mice in the three-chamber test. (C) Upper panel: Schematic diagram showing the social novelty task and representative tracking data. Lower panel: The time spent in the chambers (two-way repeated-measures analysis of variance with Sidak's test), the social interaction index (two-tailed unpaired t-test), sniffing duration (two-way repeated-measures analysis of variance with Sidak's test), and the discrimination index (two-tailed unpaired t -test with Welch's correction) in the three-chamber test (AAV-mCherry group, n = 7; AAV-hM4 group, n = 8). (D) Representative images showing c-Fos-positive cells in the LS of AAV-mCherry and AAV-hM4 mice subjected to 30 minutes of restraint. Quantitative analysis of the number of c-Fos-positive cells in the LS (two-tailed unpaired t -test; AAV-mCherry group, n = 8 slices from three mice; AAV-hM4 group, n = 9 slices from three mice). Scale bars: 100 μm at low magnification and 25 μm at high magnification. Data are expressed as mean ± SEM. * P < 0.05, ** P < 0.01, *** P < 0.001, vs. AAV-mCherry group. AAV: Adeno-associated viruses; AP: action potential; CNO: clozapine-N-oxide; DIO: double-floxed inverted open reading frame; ER: estrogen receptor; Fos: FBJ murine osteosarcoma viral oncogene homolog; GABA: gamma-aminobutyric acid; hM4: hM4D(Gi); LS: lateral septum; LV: lateral ventricle.

Article Snippet: All sections were kept at 4°C overnight in a blocking solution containing the following primary antibodies: rabbit anti-c-FBJ murine osteosarcoma viral oncogene homolog (Fos; 1:1000, CST, Danvers, MA, USA, Cat# 2250S, RRID: AB_2247211), rabbit anti-ΔFosB (1:1000, CST, Cat# 2251S, RRID: AB_2106903), rabbit anti-GABA (1:500, Sigma-Aldrich, Cat# A2052, RRID: AB_477652), and mouse anti-vesicular glutamate transporter 2 (Vglut2; 1:400, Abcam, Cambridge, UK, Cat# ab79157, RRID: AB_1603114).

Techniques: Inhibition, Expressing, Two Tailed Test

Journal: Neural Regeneration Research

Article Title: How do lateral septum projections to the ventral CA1 influence sociability?

doi: 10.4103/1673-5374.389304

Figure Lengend Snippet: Social behaviors are enhanced by re-activating socially activated LS GABA neurons. (A) Representative image showing the expression of AAV-DIO-hM3-mCherry in the LS and the experimental timeline. Illustrations were created with BioRender.com. (B) Upper panel: Schematic diagram showing the social preference task and the representative tracking activity data. Lower panel: The time spent in the chambers (two-way repeated-measures analysis of variance with Sidak's test), the social interaction index (two-tailed unpaired t -test), sniffing duration (two-way repeated-measures analysis of variance with Sidak's test), and the discrimination index (two-tailed unpaired t -test) in the three-chamber test. (C) Upper panel: Schematic diagram showing the social novelty task and the representative tracking activity. Lower panel: The time spent in the chambers (two-way repeated-measures analysis of variance with Sidak's test), the social interaction index (Mann-Whitney U test), sniffing duration (two-way repeated-measures analysis of variance with Sidak's test), and the discrimination index (Mann-Whitney U test) in the three-chamber test (AAV-mCherry group, n = 7; AAV-hM3 group, n = 8). (D) Representative images showing c-Fos-positive cells, and quantitative analysis of the number of c-Fos-positive cells in the LS (two-tailed unpaired t -test; AAV-mCherry group, n = 8 slices from three mice; AAV-hM3 group, n = 9 slices from three mice). Scale bars: 100 μm at low magnification and 25 μm at high magnification. Data are expressed as mean ± SEM. * P < 0.05, ** P < 0.01, *** P < 0.001, vs . AAV-mCherry group. AAV: Adeno-associated viruses; AP: action potential; CNO: clozapine-N-oxide; DIO: double-floxed inverted open reading frame; ER: estrogen receptor; Fos: FBJ murine osteosarcoma viral oncogene homolog; GABA: gamma-aminobutyric acid; hM3: hM3D(Gq); LS: lateral septum; LV: lateral ventricle.

Article Snippet: All sections were kept at 4°C overnight in a blocking solution containing the following primary antibodies: rabbit anti-c-FBJ murine osteosarcoma viral oncogene homolog (Fos; 1:1000, CST, Danvers, MA, USA, Cat# 2250S, RRID: AB_2247211), rabbit anti-ΔFosB (1:1000, CST, Cat# 2251S, RRID: AB_2106903), rabbit anti-GABA (1:500, Sigma-Aldrich, Cat# A2052, RRID: AB_477652), and mouse anti-vesicular glutamate transporter 2 (Vglut2; 1:400, Abcam, Cambridge, UK, Cat# ab79157, RRID: AB_1603114).

Techniques: Expressing, Activity Assay, Two Tailed Test, MANN-WHITNEY