aav2 dio vector Search Results


aav2-ef1a-dio-oxr2-sv40pa  (Virovek Inc)
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Virovek Inc aav2-ef1a-dio-oxr2-sv40pa
a , ChR2-mediated stimulation of GAD2 LHb neurons in NONs did not affect attack latency during RI (two-tailed paired t-test, n=7 biologically independent mice, t(6)=1.0, p=0.3559). b , ChR2-mediated stimulation of GAD2 LHb neurons in NONs did not affect attack duration during RI (two-tailed paired t-test, n=7 biologically independent mice, t(6)=1.0, p=0.3559). c , ChR2-mediated stimulation of orexin terminals in the LHb did not affect attack latency during RI (two-tailed paired t-test, n=6 biologically independent mice, t(5)=1.0, p=0.3632). d , ChR2-mediated stimulation of orexin terminals in the LHb did not affect attack duration during RI (two-tailed paired t-test, n=5 biologically independent mice, t(5)=1.0, p=0.3632). e , Over-expression of <t>OxR2</t> in GAD2 LHb neurons in NONs did not affect attack latency during RI (two-tailed student’s t-test, n=8 biologically independent GFP mice and n=11 biologically independent OxR2-OE mice, t(17)=0.8338, p=0.4160). f , Over-expression of OxR2 in GAD2 LHb neurons in NONs did not affect attack duration during RI (two-tailed student’s t-test, n=8 biologically independent GFP mice and n=11 biologically independent OxR2-OE mice, t(17)=0.9951. All data are expressed as mean ± SEM.
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a , ChR2-mediated stimulation of GAD2 LHb neurons in NONs did not affect attack latency during RI (two-tailed paired t-test, n=7 biologically independent mice, t(6)=1.0, p=0.3559). b , ChR2-mediated stimulation of GAD2 LHb neurons in NONs did not affect attack duration during RI (two-tailed paired t-test, n=7 biologically independent mice, t(6)=1.0, p=0.3559). c , ChR2-mediated stimulation of orexin terminals in the LHb did not affect attack latency during RI (two-tailed paired t-test, n=6 biologically independent mice, t(5)=1.0, p=0.3632). d , ChR2-mediated stimulation of orexin terminals in the LHb did not affect attack duration during RI (two-tailed paired t-test, n=5 biologically independent mice, t(5)=1.0, p=0.3632). e , Over-expression of OxR2 in GAD2 LHb neurons in NONs did not affect attack latency during RI (two-tailed student’s t-test, n=8 biologically independent GFP mice and n=11 biologically independent OxR2-OE mice, t(17)=0.8338, p=0.4160). f , Over-expression of OxR2 in GAD2 LHb neurons in NONs did not affect attack duration during RI (two-tailed student’s t-test, n=8 biologically independent GFP mice and n=11 biologically independent OxR2-OE mice, t(17)=0.9951. All data are expressed as mean ± SEM.

Journal: Nature neuroscience

Article Title: Orexin signaling in GABAergic lateral habenula neurons modulates aggressive behavior in male mice

doi: 10.1038/s41593-020-0617-7

Figure Lengend Snippet: a , ChR2-mediated stimulation of GAD2 LHb neurons in NONs did not affect attack latency during RI (two-tailed paired t-test, n=7 biologically independent mice, t(6)=1.0, p=0.3559). b , ChR2-mediated stimulation of GAD2 LHb neurons in NONs did not affect attack duration during RI (two-tailed paired t-test, n=7 biologically independent mice, t(6)=1.0, p=0.3559). c , ChR2-mediated stimulation of orexin terminals in the LHb did not affect attack latency during RI (two-tailed paired t-test, n=6 biologically independent mice, t(5)=1.0, p=0.3632). d , ChR2-mediated stimulation of orexin terminals in the LHb did not affect attack duration during RI (two-tailed paired t-test, n=5 biologically independent mice, t(5)=1.0, p=0.3632). e , Over-expression of OxR2 in GAD2 LHb neurons in NONs did not affect attack latency during RI (two-tailed student’s t-test, n=8 biologically independent GFP mice and n=11 biologically independent OxR2-OE mice, t(17)=0.8338, p=0.4160). f , Over-expression of OxR2 in GAD2 LHb neurons in NONs did not affect attack duration during RI (two-tailed student’s t-test, n=8 biologically independent GFP mice and n=11 biologically independent OxR2-OE mice, t(17)=0.9951. All data are expressed as mean ± SEM.

Article Snippet: To create an effective Cre-dependent over-expression virus for OxR2, the sequence for OxR2 was inserted into a Cre-dependent AAV2.Ef1a.DIO vector and packaged to produce AAV2-Ef1a-DIO-OxR2-SV40pA (Virovek,Inc.).

Techniques: Two Tailed Test, Over Expression

a , Representative in-situ hybridization (ISH) images showing OxR2 expression in GAD2 and vGlut2 LHb neurons, scale bar=20 μm. b , OxR2 is expressed primarily in GAD2 neurons compared to vGlut2 neurons (two-tailed student’s t-test, n=3 biologically independent mice, 1–2 slices per mouse, t(4)=15.67, p<0.0001). c , GAD2 neurons express more OxR2 mRNA than vGlut2 neurons (two-tailed student’s t-test, n=3 biologically independent mice, 1–2 slices per mouse, t(4)=15.67, p<0.0001). d , Surgical manipulations and experimental schematic for orexin bath application experiments. e , Representative traces from a GFP-positive neuron (GAD2 neuron) during baseline (top), orexin-A (middle), and washout (bottom) conditions. f , Orexin bath application increased the firing rate of GFP-positive neurons (Kruskal-Wallis one-way ANOVA with repeated measures, n=6 biologically independent mice, n=9 cells, Kruskall-Wallis statistic=7.115, p=0.0285, Dunn’s multiple comparisons baseline vs. orexin-A, adjusted p=0.0456). g , Experimental timeline for LHb qPCR experiments. h , Following 3 days of RI, AGGs expressed significantly more LHb OxR2 mRNA than NONs (3 days RI: two-tailed student’s t-test, n=7 biologically independent NON mice, n=8 biologically independent AGG mice, t(13)=2.496, p=0.0268; 1 day RI: two-tailed student’s t-test, n=9 biologically independent NON mice, n=7 biologically independent AGG mice, t(14)=0.1433, p=0.881). i , Experimental timeline for OxR2 ISH experiments. j , Following 3 days of RI, AGGs displayed increased GAD2 neuron OxR2 mRNA compared to NONs (two-tailed student’s t-test, n=5 biologically independent NON mice, n=6 biologically independent AGG mice, GAD2 neurons: t(9)6.039, p=0.0002; vGlut2 neurons: t(9)=0.6735, p=0.5175). k , Representative images of ISH for OxR2, GAD2, in AGGs and NONs following 3 days of RI, scale bar=20 μm. Experimental images were obtained from 11 independent mice with two slices imaged per mouse, with similar results obtained. *p<0.05, ***p<0.001 All data are expressed as mean ± SEM.

Journal: Nature neuroscience

Article Title: Orexin signaling in GABAergic lateral habenula neurons modulates aggressive behavior in male mice

doi: 10.1038/s41593-020-0617-7

Figure Lengend Snippet: a , Representative in-situ hybridization (ISH) images showing OxR2 expression in GAD2 and vGlut2 LHb neurons, scale bar=20 μm. b , OxR2 is expressed primarily in GAD2 neurons compared to vGlut2 neurons (two-tailed student’s t-test, n=3 biologically independent mice, 1–2 slices per mouse, t(4)=15.67, p<0.0001). c , GAD2 neurons express more OxR2 mRNA than vGlut2 neurons (two-tailed student’s t-test, n=3 biologically independent mice, 1–2 slices per mouse, t(4)=15.67, p<0.0001). d , Surgical manipulations and experimental schematic for orexin bath application experiments. e , Representative traces from a GFP-positive neuron (GAD2 neuron) during baseline (top), orexin-A (middle), and washout (bottom) conditions. f , Orexin bath application increased the firing rate of GFP-positive neurons (Kruskal-Wallis one-way ANOVA with repeated measures, n=6 biologically independent mice, n=9 cells, Kruskall-Wallis statistic=7.115, p=0.0285, Dunn’s multiple comparisons baseline vs. orexin-A, adjusted p=0.0456). g , Experimental timeline for LHb qPCR experiments. h , Following 3 days of RI, AGGs expressed significantly more LHb OxR2 mRNA than NONs (3 days RI: two-tailed student’s t-test, n=7 biologically independent NON mice, n=8 biologically independent AGG mice, t(13)=2.496, p=0.0268; 1 day RI: two-tailed student’s t-test, n=9 biologically independent NON mice, n=7 biologically independent AGG mice, t(14)=0.1433, p=0.881). i , Experimental timeline for OxR2 ISH experiments. j , Following 3 days of RI, AGGs displayed increased GAD2 neuron OxR2 mRNA compared to NONs (two-tailed student’s t-test, n=5 biologically independent NON mice, n=6 biologically independent AGG mice, GAD2 neurons: t(9)6.039, p=0.0002; vGlut2 neurons: t(9)=0.6735, p=0.5175). k , Representative images of ISH for OxR2, GAD2, in AGGs and NONs following 3 days of RI, scale bar=20 μm. Experimental images were obtained from 11 independent mice with two slices imaged per mouse, with similar results obtained. *p<0.05, ***p<0.001 All data are expressed as mean ± SEM.

Article Snippet: To create an effective Cre-dependent over-expression virus for OxR2, the sequence for OxR2 was inserted into a Cre-dependent AAV2.Ef1a.DIO vector and packaged to produce AAV2-Ef1a-DIO-OxR2-SV40pA (Virovek,Inc.).

Techniques: In Situ Hybridization, Expressing, Two Tailed Test

a , Attack latency for one day of RI in mice used for LHb qPCR, n=9 biologically independent NON mice, n=7 biologically independent AGG mice. b , Average attack latency for three days of RI in mice used for LHb qPCR, n=7 biologically independent NON mice, n=8 biologially independent AGG mice. , Average attack latency for three days of RI in mice used for LHb OxR2 ISH, n=6 biologically independent NON mice, n=6 biologically independent AGG mice. d , Representative images from OxR2 ISH in AGG and NON LHb vGlut2 neurons following RI, accompanies , scale bar = 20 μm. Notably, OxR2 expression was barely detectable in these neurons in AGGs or NONs, which is in line with our findings showing low OxR2 expression in vGlut2 neurons in – . Experimental images were taken from 12 biologically independent mice, 2 slices per mouse, with similar results obtained. All data are expressed as mean ± SEM.

Journal: Nature neuroscience

Article Title: Orexin signaling in GABAergic lateral habenula neurons modulates aggressive behavior in male mice

doi: 10.1038/s41593-020-0617-7

Figure Lengend Snippet: a , Attack latency for one day of RI in mice used for LHb qPCR, n=9 biologically independent NON mice, n=7 biologically independent AGG mice. b , Average attack latency for three days of RI in mice used for LHb qPCR, n=7 biologically independent NON mice, n=8 biologially independent AGG mice. , Average attack latency for three days of RI in mice used for LHb OxR2 ISH, n=6 biologically independent NON mice, n=6 biologically independent AGG mice. d , Representative images from OxR2 ISH in AGG and NON LHb vGlut2 neurons following RI, accompanies , scale bar = 20 μm. Notably, OxR2 expression was barely detectable in these neurons in AGGs or NONs, which is in line with our findings showing low OxR2 expression in vGlut2 neurons in – . Experimental images were taken from 12 biologically independent mice, 2 slices per mouse, with similar results obtained. All data are expressed as mean ± SEM.

Article Snippet: To create an effective Cre-dependent over-expression virus for OxR2, the sequence for OxR2 was inserted into a Cre-dependent AAV2.Ef1a.DIO vector and packaged to produce AAV2-Ef1a-DIO-OxR2-SV40pA (Virovek,Inc.).

Techniques: Expressing

a , Experimental scheme for OxR2 systemic antagonism RI experiment. b , RI test attack latency in animals treated with EMPA and vehicle (two-tailed paired t-test, n=11 biologically independent mice per group, t(10)=0.3215, p=0.758). c , RI test attack duration in animals treated with EMPA and vehicle (two-tailed paired t-test, n=11 biologically independent mice per group, t(10)=2.888, p=0.016). d , Experimental scheme for OxR2 systemic antagonism aggression CPP and locomotion experiments. e , Aggression CPP for animals treated with EMPA and vehicle (two-tailed student’s t-test, n=12 biologically independent vehicle mice and n=11 biologically independent EMPA mice, t(21)=2.885, p=0.0086). f , Locomotor activity in the open field for animals treated with EMPA and vehicle (two-tailed student’s t-test, n=12 biologically independent vehicle mice and n=11 biologically independent EMPA mice, t(21)=0.1301, p=0.8991). g , Anxiety-related behavior in the open field for animals treated with EMPA or vehicle (two-tailed student’s t-test, n=11 biologically independent mice per group, t(21)=1.134, p=0.2695) h , Representative fiber photometry traces in an animal treated with vehicle and EMPA. i , LHb GCaMP peaks during RI during vehicle and EMPA treatment (two-tailed paired t-test, n=5 biologically independent mice, t(4)=2.946, p=0.0421). *p<0.05, **p<0.01. All data are expressed as mean ± SEM.

Journal: Nature neuroscience

Article Title: Orexin signaling in GABAergic lateral habenula neurons modulates aggressive behavior in male mice

doi: 10.1038/s41593-020-0617-7

Figure Lengend Snippet: a , Experimental scheme for OxR2 systemic antagonism RI experiment. b , RI test attack latency in animals treated with EMPA and vehicle (two-tailed paired t-test, n=11 biologically independent mice per group, t(10)=0.3215, p=0.758). c , RI test attack duration in animals treated with EMPA and vehicle (two-tailed paired t-test, n=11 biologically independent mice per group, t(10)=2.888, p=0.016). d , Experimental scheme for OxR2 systemic antagonism aggression CPP and locomotion experiments. e , Aggression CPP for animals treated with EMPA and vehicle (two-tailed student’s t-test, n=12 biologically independent vehicle mice and n=11 biologically independent EMPA mice, t(21)=2.885, p=0.0086). f , Locomotor activity in the open field for animals treated with EMPA and vehicle (two-tailed student’s t-test, n=12 biologically independent vehicle mice and n=11 biologically independent EMPA mice, t(21)=0.1301, p=0.8991). g , Anxiety-related behavior in the open field for animals treated with EMPA or vehicle (two-tailed student’s t-test, n=11 biologically independent mice per group, t(21)=1.134, p=0.2695) h , Representative fiber photometry traces in an animal treated with vehicle and EMPA. i , LHb GCaMP peaks during RI during vehicle and EMPA treatment (two-tailed paired t-test, n=5 biologically independent mice, t(4)=2.946, p=0.0421). *p<0.05, **p<0.01. All data are expressed as mean ± SEM.

Article Snippet: To create an effective Cre-dependent over-expression virus for OxR2, the sequence for OxR2 was inserted into a Cre-dependent AAV2.Ef1a.DIO vector and packaged to produce AAV2-Ef1a-DIO-OxR2-SV40pA (Virovek,Inc.).

Techniques: Two Tailed Test, Activity Assay

a , >90% of neurons infected with AAV1-DIO-YFP were positive for orexin-A protein as determined by immunohistochemistry, n=3 biologically independent mice, 3 slices per mouse. b , Surgical manipulations for ChR2-mediated activation of orexin terminals in the LHb with concurrent knockdown of LHb OxR2. c , Optogenetic stimulation of orexin terminals in the LHb reduced attack latency in mice treated with the miR-scrambed virus, but not the miR-OxR2 virus (two-tailed paired t-test, miR-scrambled: n=11 biologially independent mice, t(10)=2.424, p=0.0358; miR-OxR2: n=9 biologically independent mice, t(8)=0.5281, p=0.6117). d , Optogenetic stimulation of orexin terminals in the LHb increased attack duration in mice treated with the miR-scrambled virus, but not the miR-OxR2 virus (two-tailed paired t-test, miR-scrambled: n=11 biologically independent mice, t(10)=2.260, p=0.0474; miR-OxR2: n=9 biologically independent mice, t(8)=0.8493, p=0.4204). *p<0.05. All data are expressed as mean ± SEM.

Journal: Nature neuroscience

Article Title: Orexin signaling in GABAergic lateral habenula neurons modulates aggressive behavior in male mice

doi: 10.1038/s41593-020-0617-7

Figure Lengend Snippet: a , >90% of neurons infected with AAV1-DIO-YFP were positive for orexin-A protein as determined by immunohistochemistry, n=3 biologically independent mice, 3 slices per mouse. b , Surgical manipulations for ChR2-mediated activation of orexin terminals in the LHb with concurrent knockdown of LHb OxR2. c , Optogenetic stimulation of orexin terminals in the LHb reduced attack latency in mice treated with the miR-scrambed virus, but not the miR-OxR2 virus (two-tailed paired t-test, miR-scrambled: n=11 biologially independent mice, t(10)=2.424, p=0.0358; miR-OxR2: n=9 biologically independent mice, t(8)=0.5281, p=0.6117). d , Optogenetic stimulation of orexin terminals in the LHb increased attack duration in mice treated with the miR-scrambled virus, but not the miR-OxR2 virus (two-tailed paired t-test, miR-scrambled: n=11 biologically independent mice, t(10)=2.260, p=0.0474; miR-OxR2: n=9 biologically independent mice, t(8)=0.8493, p=0.4204). *p<0.05. All data are expressed as mean ± SEM.

Article Snippet: To create an effective Cre-dependent over-expression virus for OxR2, the sequence for OxR2 was inserted into a Cre-dependent AAV2.Ef1a.DIO vector and packaged to produce AAV2-Ef1a-DIO-OxR2-SV40pA (Virovek,Inc.).

Techniques: Infection, Immunohistochemistry, Activation Assay, Two Tailed Test

a , Surgical manipulations for GAD2 neuron-specific knockdown of OxR2 in the LHb. b , Representative viral infection for GAD2 neuron-specific knockdown of OxR2 in the LHb, scale bar=300 μm. c , OxR2 knockdown in GAD2 LHb neurons increased attack latency in RI (two-tailed student’s t-test, n=12 biologically independent mice per group, t(22)=2.322, p=0.0299). d , OxR2 knockdown in GAD2 LHb neurons reduced attack duration in RI (two-tailed student’s t-test, n=12 biologically independent mice per group, t(22)=3.183, p=0.0043). e , OxR2 knockdown in GAD2 LHb neurons reduced aggression CPP (two-tailed student’s t-test, n=12 biologically independent mice per group, t(22)=2.155, p=0,0424). * p<0.05, **p<0.01. All data are expressed as mean ± SEM.

Journal: Nature neuroscience

Article Title: Orexin signaling in GABAergic lateral habenula neurons modulates aggressive behavior in male mice

doi: 10.1038/s41593-020-0617-7

Figure Lengend Snippet: a , Surgical manipulations for GAD2 neuron-specific knockdown of OxR2 in the LHb. b , Representative viral infection for GAD2 neuron-specific knockdown of OxR2 in the LHb, scale bar=300 μm. c , OxR2 knockdown in GAD2 LHb neurons increased attack latency in RI (two-tailed student’s t-test, n=12 biologically independent mice per group, t(22)=2.322, p=0.0299). d , OxR2 knockdown in GAD2 LHb neurons reduced attack duration in RI (two-tailed student’s t-test, n=12 biologically independent mice per group, t(22)=3.183, p=0.0043). e , OxR2 knockdown in GAD2 LHb neurons reduced aggression CPP (two-tailed student’s t-test, n=12 biologically independent mice per group, t(22)=2.155, p=0,0424). * p<0.05, **p<0.01. All data are expressed as mean ± SEM.

Article Snippet: To create an effective Cre-dependent over-expression virus for OxR2, the sequence for OxR2 was inserted into a Cre-dependent AAV2.Ef1a.DIO vector and packaged to produce AAV2-Ef1a-DIO-OxR2-SV40pA (Virovek,Inc.).

Techniques: Infection, Two Tailed Test

a , N2A cells treated with miR-OxR2 construct selectively reduced OxR2 expression compared to cells treated with miR-scrambled construct, but did not reduce expression of related transcripts (two-tailed student’s t-test, n=3 biologically independent plates per group, 3 replicates per plate; OxR2: t(4)=2.402, p=0.0482; OxR1: t(4)=0.2123, p=0.8423; Avpr2: t(4)=0.3686, p=0.7311; Htr3a: t(4)=1.309, p=0.2607; Drd4: t(4)=0.1925, p=0.8567; Nmur: t(4)=1.672, p=0.1699; Drd1: t(4)=0.9239, p=0.4078; Mchr1: t(4)=1.467, p=0.2163; Glpr1: t(4)=1.785, p=0.1488). b , GAD2-Cre mice injected with AAV-DIO-miR-OxR2 displayed reduced expression of OxR2 compared to mice injected with AAV-DIO-miR-scrambled as determined by ISH (two-tailed student’s t-test, n=3 mice, 2 slices per mouse, t(4)=18.44, p=0.0001). c , Representative image of GFP expression localized to GAD2 LHb neurons in GAD2-Cre mice injected with AAV-DIO-miR-OxR2, scale bar = 25 μm. Experimental images were obtained from 6 biologically independent mice, 2 slices per mouse, with similar results obtained. d , Representative images of OxR2 expression in GAD2 LHb neurons infected with AAV-DIO-miR-OxR2 or AAV-DIO-miR-scrambled, scale bar, 20 μm. *p<0.05, ***p<0.001. All data are expressed as mean ± SEM.

Journal: Nature neuroscience

Article Title: Orexin signaling in GABAergic lateral habenula neurons modulates aggressive behavior in male mice

doi: 10.1038/s41593-020-0617-7

Figure Lengend Snippet: a , N2A cells treated with miR-OxR2 construct selectively reduced OxR2 expression compared to cells treated with miR-scrambled construct, but did not reduce expression of related transcripts (two-tailed student’s t-test, n=3 biologically independent plates per group, 3 replicates per plate; OxR2: t(4)=2.402, p=0.0482; OxR1: t(4)=0.2123, p=0.8423; Avpr2: t(4)=0.3686, p=0.7311; Htr3a: t(4)=1.309, p=0.2607; Drd4: t(4)=0.1925, p=0.8567; Nmur: t(4)=1.672, p=0.1699; Drd1: t(4)=0.9239, p=0.4078; Mchr1: t(4)=1.467, p=0.2163; Glpr1: t(4)=1.785, p=0.1488). b , GAD2-Cre mice injected with AAV-DIO-miR-OxR2 displayed reduced expression of OxR2 compared to mice injected with AAV-DIO-miR-scrambled as determined by ISH (two-tailed student’s t-test, n=3 mice, 2 slices per mouse, t(4)=18.44, p=0.0001). c , Representative image of GFP expression localized to GAD2 LHb neurons in GAD2-Cre mice injected with AAV-DIO-miR-OxR2, scale bar = 25 μm. Experimental images were obtained from 6 biologically independent mice, 2 slices per mouse, with similar results obtained. d , Representative images of OxR2 expression in GAD2 LHb neurons infected with AAV-DIO-miR-OxR2 or AAV-DIO-miR-scrambled, scale bar, 20 μm. *p<0.05, ***p<0.001. All data are expressed as mean ± SEM.

Article Snippet: To create an effective Cre-dependent over-expression virus for OxR2, the sequence for OxR2 was inserted into a Cre-dependent AAV2.Ef1a.DIO vector and packaged to produce AAV2-Ef1a-DIO-OxR2-SV40pA (Virovek,Inc.).

Techniques: Construct, Expressing, Two Tailed Test, Injection, Infection

a , N2A cells treated with OxR2 over-expression construct selectively increased OxR2 expression compared to controls (two-tailed student’s t-test, n=3 biologically independent plates per group, 3 replicates per plate, OxR2: t(4)=3.939, p=0.0171; OxR1: t(4)=0.1238, p=0.9075). b , GAD2-Cre mice injected with AAV-DIO-OxR2 displayed increased expression of OxR2 compared to mice injected with AAV-DIO-GFP as determined by ISH (two-tailed student’s t-test, n=3 biologically independent mice, 2 slices per mouse, t(4)=4.417, p=0.0069) (left). Representative image of GFP expression localized to GAD2 LHb neurons in GAD2-Cre mice injected with AAV-DIO-OxR2, scale bar = 25 μm (right). Experimental images were obtained from 7 biologically independent mice, 2 slices per mouse, with similar results obtained. c , Representative images of OxR2 expression in GAD2 LHb neurons infected with AAV-DIO-OxR2 or AAV-DIO-GFP, scale bar = 25 μm. *p<0.05, **p<0.01. Experimental images were taken from 7 biologically independent mice, 2 slices per mouse, with similar results obtained. All data are expressed as mean ± SEM.

Journal: Nature neuroscience

Article Title: Orexin signaling in GABAergic lateral habenula neurons modulates aggressive behavior in male mice

doi: 10.1038/s41593-020-0617-7

Figure Lengend Snippet: a , N2A cells treated with OxR2 over-expression construct selectively increased OxR2 expression compared to controls (two-tailed student’s t-test, n=3 biologically independent plates per group, 3 replicates per plate, OxR2: t(4)=3.939, p=0.0171; OxR1: t(4)=0.1238, p=0.9075). b , GAD2-Cre mice injected with AAV-DIO-OxR2 displayed increased expression of OxR2 compared to mice injected with AAV-DIO-GFP as determined by ISH (two-tailed student’s t-test, n=3 biologically independent mice, 2 slices per mouse, t(4)=4.417, p=0.0069) (left). Representative image of GFP expression localized to GAD2 LHb neurons in GAD2-Cre mice injected with AAV-DIO-OxR2, scale bar = 25 μm (right). Experimental images were obtained from 7 biologically independent mice, 2 slices per mouse, with similar results obtained. c , Representative images of OxR2 expression in GAD2 LHb neurons infected with AAV-DIO-OxR2 or AAV-DIO-GFP, scale bar = 25 μm. *p<0.05, **p<0.01. Experimental images were taken from 7 biologically independent mice, 2 slices per mouse, with similar results obtained. All data are expressed as mean ± SEM.

Article Snippet: To create an effective Cre-dependent over-expression virus for OxR2, the sequence for OxR2 was inserted into a Cre-dependent AAV2.Ef1a.DIO vector and packaged to produce AAV2-Ef1a-DIO-OxR2-SV40pA (Virovek,Inc.).

Techniques: Over Expression, Construct, Expressing, Two Tailed Test, Injection, Infection