aav9 hsyn dio hm3d g q mcherry  (Addgene inc)

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 chemogenetic studies, either AAV-hSyn-DIO-hM3D(Gq)–mCherry (cat #44361; Addgene) or AAV-hSyn-fDIO-hM3D(Gq)-mCherry-WPREpA (cat#154868; Addgene) was used.

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

Journal: bioRxiv

Article Title: Vagal dopaminergic afferents link interoception to trigeminal pain modulation

doi: 10.64898/2026.03.27.714928

Figure Lengend Snippet: ( A ) Experimental timeline. AAV vectors were injected into the nodose ganglion (NG) 28 days before the forced mouth-opening (FMO) procedure. FMO was performed for 3 h per day across five consecutive days. Facial mechanical thresholds (von Frey), grimace scores (MGS), and open-field activity were assessed at baseline and multiple time points after FMO. C21 was administered intraperitoneally once daily for 2 days beginning on day 1 post-FMO, and conditioned place-preference (CPP) testing was conducted between days 9–13 post-FMO. ( B ) Representative confocal fluorescence images of NG showing mCherry reporter expression following viral infection, with nuclei counterstained with DAPI. ( C ) Facial withdrawal thresholds across different Cre lines and wild-type after FMO. C21 was administered intraperitoneally after von Frey testing once daily on day 1 and 2 after FMO. Cre-driver mice received nodose ganglion injection of AAV-DIO-hM3Dq and control mice received AAV-DIO-mCherry, both received intraperitoneal C21 after injection. ( D ) Facial withdrawal thresholds in DAT-Cre mice or wild type mice with or without intra-TMJ injection of C21. DAT-cre mice received nodose ganglion injection of AAV-DIO-hM3Dq, followed by intra-TMJ injection of C21 or vehicle. Data are presented as mean ± s.e.m.: *p < 0.05, **p < 0.01, ***p < 0.001 (two-way ANOVA followed by Tukey’s multiple-comparisons test). ( E ) Representative facial images of WT and DAT-Cre mice before and after FMO with or without C21 treatment. ( F ) Quantification of spontaneous pain using the Mouse Grimace Scale (MGS). DAT-Cre + C21 mice exhibited significantly reduced MGS scores on day 5 and day 7 post-FMO compared to control groups. Data are mean ± s.e.m.; **p < 0.01, ***p < 0.001, ###p < 0.001 (two-way ANOVA followed by Tukey’s multiple-comparisons test).

Article Snippet: For selective activation of dopaminergic vagal neurons, AAV-DIO-hM3Dq (#44361; Addgene) or AAV-DIO-mCherry (control) (#50459; Addgene) was injected unilaterally into the right nodose ganglion of Cre-driver mice.

Techniques: Injection, Activity Assay, Conditioned Place Preference, Fluorescence, Expressing, Infection, Control