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aav hsynflex mgfp 2a synaptophysin mruby  (Addgene inc)


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    Addgene inc aav hsynflex mgfp 2a synaptophysin mruby
    Aav Hsynflex Mgfp 2a Synaptophysin Mruby, supplied by Addgene inc, used in various techniques. Bioz Stars score: 94/100, based on 63 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/aav+flex/pAAV+hSyn+FLEx+mGFP-2A-Synaptophysin-mRuby+(Plasmid+%2371760)/pm41746362-225-9-12
    Average 94 stars, based on 63 article reviews
    aav hsynflex mgfp 2a synaptophysin mruby - by Bioz Stars, 2026-09
    94/100 stars

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    Related Articles

    Bioprocessing:

    Article Title: Peptidergic and functional delineation of the Edinger-Westphal nucleus
    Article Snippet: AAV1-EF1α-DIO-LCK-APEX2-P2A-EGFP was packaged from AAV-EF1a-DIO-LckAPEX-P2A-EGFP plasmid, which we have deposited at Addgene (plasmid # 182826). .. AAV1-CBA-FLEX-ChR2-mCherry was packaged from AAV-FLEX-rev-ChR2(H134R)-mCherry plasmid, which was a gift from Scott Sternson (Addgene plasmid #18916). .. AAVrg-CAG-FLEX-EGFP was packaged from AAV pCAG-FLEX-EGFP-WPRE, which was a gift from Hongkui Zeng (Addgene viral prep # 51502-AAVrg).

    Article Title: Molecularly Defined Circuitry Reveals Input-Output Segregation in Deep Layers of the Medial Entorhinal Cortex
    Article Snippet: .. For viral transduction of entorhinal neurons, ∼200 nl of one of the following viruses was injected through a glass pipette: AAV-FLEX-rev-ChR2mCherry, which expresses ChR2-mCherry from a CAG promoter ( , AddGene 18916) (titer: 2.2 × 10 14 cp/ml, measured by qPCR. cp stands for capsid particle); AAV-FLEX-GFP, which expresses GFP from a CBA promoter ( ) (titer: 1.5 × 10 12 cp/ml); or AAV-FLEX-synaptophysin-eGFP, which expresses synaptophysin-eGFP from a CBA promoter (modified from ) (titer: 1.2 × 10 12 cp/ml). ..

    Article Title: Peptidergic modulation of fear responses by the Edinger-Westphal nucleus
    Article Snippet: For fiber photometry, 200-250 nl of AAV1-CAG-FLEX-GCaMP6s-WPRE ( ) (University of Pennsylvania Vector Core, 4 x 10 12 to 1 x 10 13 gc/ml) was injected at - 3.5 and -3.1 D/V at a rate of 100 nl/min. .. AAV1-CBA-FLEX-ChR2-mCherry was packaged from AAV-FLEX-rev-ChR2(H134R)-mCherry plasmid, which was a gift from Scott Sternson (Addgene plasmid #18916). .. AAV1-CBA-DIO-rM3Ds-mCherry-WPRE was packaged from plasmid pAAV-hSyn-DIO-rM3D(Gs)-mCherry, which was a gift from Bryan Roth (Addgene plasmid #50458).

    Article Title: Cell-type-specific role for nucleus accumbens neuroligin-2 in depression and stress susceptibility
    Article Snippet: Short hairpin RNA sequences were then used to create microRNA oligos using the BLOCK-iTTM Pol II miR RNAi Expression Vector Kit (K4936-00; Invitrogen). miR constructs were next subcloned into a bicistronic AAV-IRES-GFP vector (VPK-418; Cell Biolabs, Inc.). .. To construct a Cre-dependent version, we modified an AAV-FLEX-rev-ChR2-Tdtomato vector (#18917; Addgene), using the Gibson assembly method (Cat# 2611; New England Biolabs). .. AAV constructs were validated in Neuro2A cells (ATCC), using common procedures before being packaged into high-titer viral particles by the University of North Carolina at Chapel Hill Gene Therapy Center (UNC Vector Core).

    Article Title: A Novel Next-Generation Sequencing and Analysis Platform to Assess the Identity of Recombinant Adeno-Associated Viral Preparations from Viral DNA Extracts
    Article Snippet: .. The following plasmids were used to prepare the rAAV described in this study; pAAV-EF1a-double floxed-hChR2(H134R)-EYFP-WPRE-HGHpA (Addgene 20298, a gift of Karl Deisseroth, unpublished) AAV-Cre-GFP (Addgene 68544, a gift of Eric Nestler), AAV-EF1a-DIO-GCaMP6s-P2A-nls-dTomato (Addgene 51082, a gift of Jonathan Ting), AAV-FLEX-rev-ChR2(H134R)-mCherry (Addgene 18916, a gift from Scott Sternson), rAAV2-retro helper (Addgene 81070, a gift of Alla Karpova), pAAV-hSyn-DIO-hM3D(Gq)-mCherry (Addgene 44361, a gift of Bryan Roth), pAAV-hSyn-DIO-hM4D(Gi)-mCherry (Addgene 44362, a gift of Bryan Roth), pAAV-hSyn-hM3D(Gq)-mCherry (Addgene 50474, a gift of Bryan Roth, unpublished), pAAV-hSyn-EGFP(Addgene 50465, a gift of Bryan Roth, unpublished), pAAV-CAG-GFP (Addgene 37825, a gift from Edward Boyden, unpublished), pAAV-CAG-tdTomato (Addgene 59462, a gift of Edward Boyden, unpublished), and pUCmini-iCAP-PHP.eB (Addgene 103005, a gift of Viviana Gradinaru). ..

    Article Title: Stellate Cells in the Medial Entorhinal Cortex Are Required for Spatial Learning
    Article Snippet: .. Viruses used where AAV-TeLC-GFP and AAV-GFP (obtained from Peer Wulf, Kiel University)(Murray et al., 2011), and AAV-FLEX-rev-ChR2(H134R)-mCherry (Addgene plasmid #18916). ..

    Article Title: Stellate Cells in the Medial Entorhinal Cortex Are Required for Spatial Learning
    Article Snippet: .. Viruses used where AAV-TeLC-GFP and AAV-GFP (obtained from Peer Wulf, Kiel University)(Murray et al., 2011), and AAV-FLEX-rev-ChR2(H134R)-mCherry (Addgene plasmid #18916). ..

    Plasmid Preparation:

    Article Title: Peptidergic and functional delineation of the Edinger-Westphal nucleus
    Article Snippet: AAV1-EF1α-DIO-LCK-APEX2-P2A-EGFP was packaged from AAV-EF1a-DIO-LckAPEX-P2A-EGFP plasmid, which we have deposited at Addgene (plasmid # 182826). .. AAV1-CBA-FLEX-ChR2-mCherry was packaged from AAV-FLEX-rev-ChR2(H134R)-mCherry plasmid, which was a gift from Scott Sternson (Addgene plasmid #18916). .. AAVrg-CAG-FLEX-EGFP was packaged from AAV pCAG-FLEX-EGFP-WPRE, which was a gift from Hongkui Zeng (Addgene viral prep # 51502-AAVrg).

    Article Title: Peptidergic modulation of fear responses by the Edinger-Westphal nucleus
    Article Snippet: For fiber photometry, 200-250 nl of AAV1-CAG-FLEX-GCaMP6s-WPRE ( ) (University of Pennsylvania Vector Core, 4 x 10 12 to 1 x 10 13 gc/ml) was injected at - 3.5 and -3.1 D/V at a rate of 100 nl/min. .. AAV1-CBA-FLEX-ChR2-mCherry was packaged from AAV-FLEX-rev-ChR2(H134R)-mCherry plasmid, which was a gift from Scott Sternson (Addgene plasmid #18916). .. AAV1-CBA-DIO-rM3Ds-mCherry-WPRE was packaged from plasmid pAAV-hSyn-DIO-rM3D(Gs)-mCherry, which was a gift from Bryan Roth (Addgene plasmid #50458).

    Article Title: Cell-type-specific role for nucleus accumbens neuroligin-2 in depression and stress susceptibility
    Article Snippet: Short hairpin RNA sequences were then used to create microRNA oligos using the BLOCK-iTTM Pol II miR RNAi Expression Vector Kit (K4936-00; Invitrogen). miR constructs were next subcloned into a bicistronic AAV-IRES-GFP vector (VPK-418; Cell Biolabs, Inc.). .. To construct a Cre-dependent version, we modified an AAV-FLEX-rev-ChR2-Tdtomato vector (#18917; Addgene), using the Gibson assembly method (Cat# 2611; New England Biolabs). .. AAV constructs were validated in Neuro2A cells (ATCC), using common procedures before being packaged into high-titer viral particles by the University of North Carolina at Chapel Hill Gene Therapy Center (UNC Vector Core).

    Article Title: Stellate Cells in the Medial Entorhinal Cortex Are Required for Spatial Learning
    Article Snippet: .. Viruses used where AAV-TeLC-GFP and AAV-GFP (obtained from Peer Wulf, Kiel University)(Murray et al., 2011), and AAV-FLEX-rev-ChR2(H134R)-mCherry (Addgene plasmid #18916). ..

    Article Title: Stellate Cells in the Medial Entorhinal Cortex Are Required for Spatial Learning
    Article Snippet: .. Viruses used where AAV-TeLC-GFP and AAV-GFP (obtained from Peer Wulf, Kiel University)(Murray et al., 2011), and AAV-FLEX-rev-ChR2(H134R)-mCherry (Addgene plasmid #18916). ..

    Transduction:

    Article Title: Molecularly Defined Circuitry Reveals Input-Output Segregation in Deep Layers of the Medial Entorhinal Cortex
    Article Snippet: .. For viral transduction of entorhinal neurons, ∼200 nl of one of the following viruses was injected through a glass pipette: AAV-FLEX-rev-ChR2mCherry, which expresses ChR2-mCherry from a CAG promoter ( , AddGene 18916) (titer: 2.2 × 10 14 cp/ml, measured by qPCR. cp stands for capsid particle); AAV-FLEX-GFP, which expresses GFP from a CBA promoter ( ) (titer: 1.5 × 10 12 cp/ml); or AAV-FLEX-synaptophysin-eGFP, which expresses synaptophysin-eGFP from a CBA promoter (modified from ) (titer: 1.2 × 10 12 cp/ml). ..

    Injection:

    Article Title: Molecularly Defined Circuitry Reveals Input-Output Segregation in Deep Layers of the Medial Entorhinal Cortex
    Article Snippet: .. For viral transduction of entorhinal neurons, ∼200 nl of one of the following viruses was injected through a glass pipette: AAV-FLEX-rev-ChR2mCherry, which expresses ChR2-mCherry from a CAG promoter ( , AddGene 18916) (titer: 2.2 × 10 14 cp/ml, measured by qPCR. cp stands for capsid particle); AAV-FLEX-GFP, which expresses GFP from a CBA promoter ( ) (titer: 1.5 × 10 12 cp/ml); or AAV-FLEX-synaptophysin-eGFP, which expresses synaptophysin-eGFP from a CBA promoter (modified from ) (titer: 1.2 × 10 12 cp/ml). ..

    Transferring:

    Article Title: Molecularly Defined Circuitry Reveals Input-Output Segregation in Deep Layers of the Medial Entorhinal Cortex
    Article Snippet: .. For viral transduction of entorhinal neurons, ∼200 nl of one of the following viruses was injected through a glass pipette: AAV-FLEX-rev-ChR2mCherry, which expresses ChR2-mCherry from a CAG promoter ( , AddGene 18916) (titer: 2.2 × 10 14 cp/ml, measured by qPCR. cp stands for capsid particle); AAV-FLEX-GFP, which expresses GFP from a CBA promoter ( ) (titer: 1.5 × 10 12 cp/ml); or AAV-FLEX-synaptophysin-eGFP, which expresses synaptophysin-eGFP from a CBA promoter (modified from ) (titer: 1.2 × 10 12 cp/ml). ..

    Real-time Polymerase Chain Reaction:

    Article Title: Molecularly Defined Circuitry Reveals Input-Output Segregation in Deep Layers of the Medial Entorhinal Cortex
    Article Snippet: .. For viral transduction of entorhinal neurons, ∼200 nl of one of the following viruses was injected through a glass pipette: AAV-FLEX-rev-ChR2mCherry, which expresses ChR2-mCherry from a CAG promoter ( , AddGene 18916) (titer: 2.2 × 10 14 cp/ml, measured by qPCR. cp stands for capsid particle); AAV-FLEX-GFP, which expresses GFP from a CBA promoter ( ) (titer: 1.5 × 10 12 cp/ml); or AAV-FLEX-synaptophysin-eGFP, which expresses synaptophysin-eGFP from a CBA promoter (modified from ) (titer: 1.2 × 10 12 cp/ml). ..

    Crocin Bleaching Assay:

    Article Title: Molecularly Defined Circuitry Reveals Input-Output Segregation in Deep Layers of the Medial Entorhinal Cortex
    Article Snippet: .. For viral transduction of entorhinal neurons, ∼200 nl of one of the following viruses was injected through a glass pipette: AAV-FLEX-rev-ChR2mCherry, which expresses ChR2-mCherry from a CAG promoter ( , AddGene 18916) (titer: 2.2 × 10 14 cp/ml, measured by qPCR. cp stands for capsid particle); AAV-FLEX-GFP, which expresses GFP from a CBA promoter ( ) (titer: 1.5 × 10 12 cp/ml); or AAV-FLEX-synaptophysin-eGFP, which expresses synaptophysin-eGFP from a CBA promoter (modified from ) (titer: 1.2 × 10 12 cp/ml). ..

    Modification:

    Article Title: Molecularly Defined Circuitry Reveals Input-Output Segregation in Deep Layers of the Medial Entorhinal Cortex
    Article Snippet: .. For viral transduction of entorhinal neurons, ∼200 nl of one of the following viruses was injected through a glass pipette: AAV-FLEX-rev-ChR2mCherry, which expresses ChR2-mCherry from a CAG promoter ( , AddGene 18916) (titer: 2.2 × 10 14 cp/ml, measured by qPCR. cp stands for capsid particle); AAV-FLEX-GFP, which expresses GFP from a CBA promoter ( ) (titer: 1.5 × 10 12 cp/ml); or AAV-FLEX-synaptophysin-eGFP, which expresses synaptophysin-eGFP from a CBA promoter (modified from ) (titer: 1.2 × 10 12 cp/ml). ..

    Article Title: Cell-type-specific role for nucleus accumbens neuroligin-2 in depression and stress susceptibility
    Article Snippet: Short hairpin RNA sequences were then used to create microRNA oligos using the BLOCK-iTTM Pol II miR RNAi Expression Vector Kit (K4936-00; Invitrogen). miR constructs were next subcloned into a bicistronic AAV-IRES-GFP vector (VPK-418; Cell Biolabs, Inc.). .. To construct a Cre-dependent version, we modified an AAV-FLEX-rev-ChR2-Tdtomato vector (#18917; Addgene), using the Gibson assembly method (Cat# 2611; New England Biolabs). .. AAV constructs were validated in Neuro2A cells (ATCC), using common procedures before being packaged into high-titer viral particles by the University of North Carolina at Chapel Hill Gene Therapy Center (UNC Vector Core).

    Construct:

    Article Title: Cell-type-specific role for nucleus accumbens neuroligin-2 in depression and stress susceptibility
    Article Snippet: Short hairpin RNA sequences were then used to create microRNA oligos using the BLOCK-iTTM Pol II miR RNAi Expression Vector Kit (K4936-00; Invitrogen). miR constructs were next subcloned into a bicistronic AAV-IRES-GFP vector (VPK-418; Cell Biolabs, Inc.). .. To construct a Cre-dependent version, we modified an AAV-FLEX-rev-ChR2-Tdtomato vector (#18917; Addgene), using the Gibson assembly method (Cat# 2611; New England Biolabs). .. AAV constructs were validated in Neuro2A cells (ATCC), using common procedures before being packaged into high-titer viral particles by the University of North Carolina at Chapel Hill Gene Therapy Center (UNC Vector Core).



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    BNST–PVN-RVLM neural signaling mediates IL-1β–induced changes in heart rate and IL-6. (A) Schematic for anterograde tracing of axonal projections and terminals from PBS-TRAPed or IL-1β–TRAPed BNST <t>neurons.</t> <t>The</t> <t>AAV-hSyn-FLEx-mGFP-2A-synaptophysin-mRuby</t> was injected into the BNST of TRAP2 mice. (B) Representative image for GFP + axons and mRuby + terminals in the PVN of PBS-TRAPed or IL-1β–TRAPed mice. The rightmost panel shows a higher-magnification view of the PVN in IL-1β–TRAPed mice. Arrowheads show regions with co-localization of mRuby and EYFP. Scale bars, 100 μm. (C) Schematic for anterograde tracing of IL-1β–TRAPed BNST neurons connected with PVN neurons. The AAV-pEF1a-DIO-FLPo-WPRE-hGHpA was injected into the BNST, and the AAV-Ef1a-fDIO-EYFP was injected into the PVN of TRAP2 mice. (D) Representative image for EYFP expression in the PVN, RVLM, and NTS. Arrowheads show neurons, and arrows show axonal projections with expression of EYFP. Scale bar, 100 μm for the PVN and RVLM. Scale bar, 200 μm for the NTS. (E) c-Fos expression in the RVLM after reactivation with saline as a control or CNO of IL-1β–responsive BNST neurons. Scale bar, 100 μm. (F) Schematic for activating the BNST–PVN neural pathway. The AAV-pEF1a-DIO-FLPo-WPRE-hGHpA was injected into the BNST, and the AAV-hSyn-fDIO-hM3D(Gq)-mCherry-WPREpA was injected into the PVN of TRAP2 mice. (G) Representative image of PVN showing Gq-DREADD-mCherry–expressing cells (red). Scale bar, 100 μm. (H) Serum IL-6 levels at 2 h after reactivation with saline as a control or CNO of the BNST–PVN neuronal pathway. Data are represented as individual mouse data points pooled from two independent experiments. Unpaired t test. (I) ΔHR for 60 min after reactivation of the BNST–PVN neuronal pathway: saline (black) or CNO (red) (saline, n = 7 mice; CNO, n = 10 mice, mixed-effects analysis with Šidák correction). (J) AUC of ΔHR after reactivation. Data are represented as individual mouse data points pooled from two independent experiments. Unpaired t test. (K) Serum corticosterone levels at 2 h after reactivation of the BNST–PVN neuronal pathway. Data are represented as individual mouse data points pooled from two independent experiments. Unpaired t test. *P < 0.05; **P < 0.01.
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    BNST–PVN-RVLM neural signaling mediates IL-1β–induced changes in heart rate and IL-6. (A) Schematic for anterograde tracing of axonal projections and terminals from PBS-TRAPed or IL-1β–TRAPed BNST <t>neurons.</t> <t>The</t> <t>AAV-hSyn-FLEx-mGFP-2A-synaptophysin-mRuby</t> was injected into the BNST of TRAP2 mice. (B) Representative image for GFP + axons and mRuby + terminals in the PVN of PBS-TRAPed or IL-1β–TRAPed mice. The rightmost panel shows a higher-magnification view of the PVN in IL-1β–TRAPed mice. Arrowheads show regions with co-localization of mRuby and EYFP. Scale bars, 100 μm. (C) Schematic for anterograde tracing of IL-1β–TRAPed BNST neurons connected with PVN neurons. The AAV-pEF1a-DIO-FLPo-WPRE-hGHpA was injected into the BNST, and the AAV-Ef1a-fDIO-EYFP was injected into the PVN of TRAP2 mice. (D) Representative image for EYFP expression in the PVN, RVLM, and NTS. Arrowheads show neurons, and arrows show axonal projections with expression of EYFP. Scale bar, 100 μm for the PVN and RVLM. Scale bar, 200 μm for the NTS. (E) c-Fos expression in the RVLM after reactivation with saline as a control or CNO of IL-1β–responsive BNST neurons. Scale bar, 100 μm. (F) Schematic for activating the BNST–PVN neural pathway. The AAV-pEF1a-DIO-FLPo-WPRE-hGHpA was injected into the BNST, and the AAV-hSyn-fDIO-hM3D(Gq)-mCherry-WPREpA was injected into the PVN of TRAP2 mice. (G) Representative image of PVN showing Gq-DREADD-mCherry–expressing cells (red). Scale bar, 100 μm. (H) Serum IL-6 levels at 2 h after reactivation with saline as a control or CNO of the BNST–PVN neuronal pathway. Data are represented as individual mouse data points pooled from two independent experiments. Unpaired t test. (I) ΔHR for 60 min after reactivation of the BNST–PVN neuronal pathway: saline (black) or CNO (red) (saline, n = 7 mice; CNO, n = 10 mice, mixed-effects analysis with Šidák correction). (J) AUC of ΔHR after reactivation. Data are represented as individual mouse data points pooled from two independent experiments. Unpaired t test. (K) Serum corticosterone levels at 2 h after reactivation of the BNST–PVN neuronal pathway. Data are represented as individual mouse data points pooled from two independent experiments. Unpaired t test. *P < 0.05; **P < 0.01.
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    BNST–PVN-RVLM neural signaling mediates IL-1β–induced changes in heart rate and IL-6. (A) Schematic for anterograde tracing of axonal projections and terminals from PBS-TRAPed or IL-1β–TRAPed BNST <t>neurons.</t> <t>The</t> <t>AAV-hSyn-FLEx-mGFP-2A-synaptophysin-mRuby</t> was injected into the BNST of TRAP2 mice. (B) Representative image for GFP + axons and mRuby + terminals in the PVN of PBS-TRAPed or IL-1β–TRAPed mice. The rightmost panel shows a higher-magnification view of the PVN in IL-1β–TRAPed mice. Arrowheads show regions with co-localization of mRuby and EYFP. Scale bars, 100 μm. (C) Schematic for anterograde tracing of IL-1β–TRAPed BNST neurons connected with PVN neurons. The AAV-pEF1a-DIO-FLPo-WPRE-hGHpA was injected into the BNST, and the AAV-Ef1a-fDIO-EYFP was injected into the PVN of TRAP2 mice. (D) Representative image for EYFP expression in the PVN, RVLM, and NTS. Arrowheads show neurons, and arrows show axonal projections with expression of EYFP. Scale bar, 100 μm for the PVN and RVLM. Scale bar, 200 μm for the NTS. (E) c-Fos expression in the RVLM after reactivation with saline as a control or CNO of IL-1β–responsive BNST neurons. Scale bar, 100 μm. (F) Schematic for activating the BNST–PVN neural pathway. The AAV-pEF1a-DIO-FLPo-WPRE-hGHpA was injected into the BNST, and the AAV-hSyn-fDIO-hM3D(Gq)-mCherry-WPREpA was injected into the PVN of TRAP2 mice. (G) Representative image of PVN showing Gq-DREADD-mCherry–expressing cells (red). Scale bar, 100 μm. (H) Serum IL-6 levels at 2 h after reactivation with saline as a control or CNO of the BNST–PVN neuronal pathway. Data are represented as individual mouse data points pooled from two independent experiments. Unpaired t test. (I) ΔHR for 60 min after reactivation of the BNST–PVN neuronal pathway: saline (black) or CNO (red) (saline, n = 7 mice; CNO, n = 10 mice, mixed-effects analysis with Šidák correction). (J) AUC of ΔHR after reactivation. Data are represented as individual mouse data points pooled from two independent experiments. Unpaired t test. (K) Serum corticosterone levels at 2 h after reactivation of the BNST–PVN neuronal pathway. Data are represented as individual mouse data points pooled from two independent experiments. Unpaired t test. *P < 0.05; **P < 0.01.
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    BNST–PVN-RVLM neural signaling mediates IL-1β–induced changes in heart rate and IL-6. (A) Schematic for anterograde tracing of axonal projections and terminals from PBS-TRAPed or IL-1β–TRAPed BNST <t>neurons.</t> <t>The</t> <t>AAV-hSyn-FLEx-mGFP-2A-synaptophysin-mRuby</t> was injected into the BNST of TRAP2 mice. (B) Representative image for GFP + axons and mRuby + terminals in the PVN of PBS-TRAPed or IL-1β–TRAPed mice. The rightmost panel shows a higher-magnification view of the PVN in IL-1β–TRAPed mice. Arrowheads show regions with co-localization of mRuby and EYFP. Scale bars, 100 μm. (C) Schematic for anterograde tracing of IL-1β–TRAPed BNST neurons connected with PVN neurons. The AAV-pEF1a-DIO-FLPo-WPRE-hGHpA was injected into the BNST, and the AAV-Ef1a-fDIO-EYFP was injected into the PVN of TRAP2 mice. (D) Representative image for EYFP expression in the PVN, RVLM, and NTS. Arrowheads show neurons, and arrows show axonal projections with expression of EYFP. Scale bar, 100 μm for the PVN and RVLM. Scale bar, 200 μm for the NTS. (E) c-Fos expression in the RVLM after reactivation with saline as a control or CNO of IL-1β–responsive BNST neurons. Scale bar, 100 μm. (F) Schematic for activating the BNST–PVN neural pathway. The AAV-pEF1a-DIO-FLPo-WPRE-hGHpA was injected into the BNST, and the AAV-hSyn-fDIO-hM3D(Gq)-mCherry-WPREpA was injected into the PVN of TRAP2 mice. (G) Representative image of PVN showing Gq-DREADD-mCherry–expressing cells (red). Scale bar, 100 μm. (H) Serum IL-6 levels at 2 h after reactivation with saline as a control or CNO of the BNST–PVN neuronal pathway. Data are represented as individual mouse data points pooled from two independent experiments. Unpaired t test. (I) ΔHR for 60 min after reactivation of the BNST–PVN neuronal pathway: saline (black) or CNO (red) (saline, n = 7 mice; CNO, n = 10 mice, mixed-effects analysis with Šidák correction). (J) AUC of ΔHR after reactivation. Data are represented as individual mouse data points pooled from two independent experiments. Unpaired t test. (K) Serum corticosterone levels at 2 h after reactivation of the BNST–PVN neuronal pathway. Data are represented as individual mouse data points pooled from two independent experiments. Unpaired t test. *P < 0.05; **P < 0.01.
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    BNST–PVN-RVLM neural signaling mediates IL-1β–induced changes in heart rate and IL-6. (A) Schematic for anterograde tracing of axonal projections and terminals from PBS-TRAPed or IL-1β–TRAPed BNST <t>neurons.</t> <t>The</t> <t>AAV-hSyn-FLEx-mGFP-2A-synaptophysin-mRuby</t> was injected into the BNST of TRAP2 mice. (B) Representative image for GFP + axons and mRuby + terminals in the PVN of PBS-TRAPed or IL-1β–TRAPed mice. The rightmost panel shows a higher-magnification view of the PVN in IL-1β–TRAPed mice. Arrowheads show regions with co-localization of mRuby and EYFP. Scale bars, 100 μm. (C) Schematic for anterograde tracing of IL-1β–TRAPed BNST neurons connected with PVN neurons. The AAV-pEF1a-DIO-FLPo-WPRE-hGHpA was injected into the BNST, and the AAV-Ef1a-fDIO-EYFP was injected into the PVN of TRAP2 mice. (D) Representative image for EYFP expression in the PVN, RVLM, and NTS. Arrowheads show neurons, and arrows show axonal projections with expression of EYFP. Scale bar, 100 μm for the PVN and RVLM. Scale bar, 200 μm for the NTS. (E) c-Fos expression in the RVLM after reactivation with saline as a control or CNO of IL-1β–responsive BNST neurons. Scale bar, 100 μm. (F) Schematic for activating the BNST–PVN neural pathway. The AAV-pEF1a-DIO-FLPo-WPRE-hGHpA was injected into the BNST, and the AAV-hSyn-fDIO-hM3D(Gq)-mCherry-WPREpA was injected into the PVN of TRAP2 mice. (G) Representative image of PVN showing Gq-DREADD-mCherry–expressing cells (red). Scale bar, 100 μm. (H) Serum IL-6 levels at 2 h after reactivation with saline as a control or CNO of the BNST–PVN neuronal pathway. Data are represented as individual mouse data points pooled from two independent experiments. Unpaired t test. (I) ΔHR for 60 min after reactivation of the BNST–PVN neuronal pathway: saline (black) or CNO (red) (saline, n = 7 mice; CNO, n = 10 mice, mixed-effects analysis with Šidák correction). (J) AUC of ΔHR after reactivation. Data are represented as individual mouse data points pooled from two independent experiments. Unpaired t test. (K) Serum corticosterone levels at 2 h after reactivation of the BNST–PVN neuronal pathway. Data are represented as individual mouse data points pooled from two independent experiments. Unpaired t test. *P < 0.05; **P < 0.01.
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    Image Search Results


    BNST–PVN-RVLM neural signaling mediates IL-1β–induced changes in heart rate and IL-6. (A) Schematic for anterograde tracing of axonal projections and terminals from PBS-TRAPed or IL-1β–TRAPed BNST neurons. The AAV-hSyn-FLEx-mGFP-2A-synaptophysin-mRuby was injected into the BNST of TRAP2 mice. (B) Representative image for GFP + axons and mRuby + terminals in the PVN of PBS-TRAPed or IL-1β–TRAPed mice. The rightmost panel shows a higher-magnification view of the PVN in IL-1β–TRAPed mice. Arrowheads show regions with co-localization of mRuby and EYFP. Scale bars, 100 μm. (C) Schematic for anterograde tracing of IL-1β–TRAPed BNST neurons connected with PVN neurons. The AAV-pEF1a-DIO-FLPo-WPRE-hGHpA was injected into the BNST, and the AAV-Ef1a-fDIO-EYFP was injected into the PVN of TRAP2 mice. (D) Representative image for EYFP expression in the PVN, RVLM, and NTS. Arrowheads show neurons, and arrows show axonal projections with expression of EYFP. Scale bar, 100 μm for the PVN and RVLM. Scale bar, 200 μm for the NTS. (E) c-Fos expression in the RVLM after reactivation with saline as a control or CNO of IL-1β–responsive BNST neurons. Scale bar, 100 μm. (F) Schematic for activating the BNST–PVN neural pathway. The AAV-pEF1a-DIO-FLPo-WPRE-hGHpA was injected into the BNST, and the AAV-hSyn-fDIO-hM3D(Gq)-mCherry-WPREpA was injected into the PVN of TRAP2 mice. (G) Representative image of PVN showing Gq-DREADD-mCherry–expressing cells (red). Scale bar, 100 μm. (H) Serum IL-6 levels at 2 h after reactivation with saline as a control or CNO of the BNST–PVN neuronal pathway. Data are represented as individual mouse data points pooled from two independent experiments. Unpaired t test. (I) ΔHR for 60 min after reactivation of the BNST–PVN neuronal pathway: saline (black) or CNO (red) (saline, n = 7 mice; CNO, n = 10 mice, mixed-effects analysis with Šidák correction). (J) AUC of ΔHR after reactivation. Data are represented as individual mouse data points pooled from two independent experiments. Unpaired t test. (K) Serum corticosterone levels at 2 h after reactivation of the BNST–PVN neuronal pathway. Data are represented as individual mouse data points pooled from two independent experiments. Unpaired t test. *P < 0.05; **P < 0.01.

    Journal: The Journal of Experimental Medicine

    Article Title: Central neurons encode interleukin-1β signals and mediate stress-induced inflammation

    doi: 10.1084/jem.20252000

    Figure Lengend Snippet: BNST–PVN-RVLM neural signaling mediates IL-1β–induced changes in heart rate and IL-6. (A) Schematic for anterograde tracing of axonal projections and terminals from PBS-TRAPed or IL-1β–TRAPed BNST neurons. The AAV-hSyn-FLEx-mGFP-2A-synaptophysin-mRuby was injected into the BNST of TRAP2 mice. (B) Representative image for GFP + axons and mRuby + terminals in the PVN of PBS-TRAPed or IL-1β–TRAPed mice. The rightmost panel shows a higher-magnification view of the PVN in IL-1β–TRAPed mice. Arrowheads show regions with co-localization of mRuby and EYFP. Scale bars, 100 μm. (C) Schematic for anterograde tracing of IL-1β–TRAPed BNST neurons connected with PVN neurons. The AAV-pEF1a-DIO-FLPo-WPRE-hGHpA was injected into the BNST, and the AAV-Ef1a-fDIO-EYFP was injected into the PVN of TRAP2 mice. (D) Representative image for EYFP expression in the PVN, RVLM, and NTS. Arrowheads show neurons, and arrows show axonal projections with expression of EYFP. Scale bar, 100 μm for the PVN and RVLM. Scale bar, 200 μm for the NTS. (E) c-Fos expression in the RVLM after reactivation with saline as a control or CNO of IL-1β–responsive BNST neurons. Scale bar, 100 μm. (F) Schematic for activating the BNST–PVN neural pathway. The AAV-pEF1a-DIO-FLPo-WPRE-hGHpA was injected into the BNST, and the AAV-hSyn-fDIO-hM3D(Gq)-mCherry-WPREpA was injected into the PVN of TRAP2 mice. (G) Representative image of PVN showing Gq-DREADD-mCherry–expressing cells (red). Scale bar, 100 μm. (H) Serum IL-6 levels at 2 h after reactivation with saline as a control or CNO of the BNST–PVN neuronal pathway. Data are represented as individual mouse data points pooled from two independent experiments. Unpaired t test. (I) ΔHR for 60 min after reactivation of the BNST–PVN neuronal pathway: saline (black) or CNO (red) (saline, n = 7 mice; CNO, n = 10 mice, mixed-effects analysis with Šidák correction). (J) AUC of ΔHR after reactivation. Data are represented as individual mouse data points pooled from two independent experiments. Unpaired t test. (K) Serum corticosterone levels at 2 h after reactivation of the BNST–PVN neuronal pathway. Data are represented as individual mouse data points pooled from two independent experiments. Unpaired t test. *P < 0.05; **P < 0.01.

    Article Snippet: For the tracing studies, either AAV-hSyn-DIO-EGFP (cat #50457; Addgene), AAV-hSyn-FLEx-mGFP-2A-Synaptophysin-mRuby (cat# 71760; Addgene), AAV-Ef1a-fDIO-EYFP (cat# 55641; Addgene), or AAV pEF1a-DIO-FLPo-WPRE-hGHpA (cat# 87306; Addgene) was utilized.

    Techniques: Anterograde Tracing, Injection, Expressing, Saline, Control