aav dio hm3dq  (Addgene inc)

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

Journal: bioRxiv

Article Title: Phenotype-driven screening reveals a causal role for the cortex in pupil control

doi: 10.64898/2026.03.17.712501

Figure Lengend Snippet: A: Experimental design for cortex-restricted chemogenetic activation. An expression plasmid for hM3Dq-mCherry was in utero electroporated unilaterally to the dorsolateral cortex of ICR wild-type embryos at E12.5-15.5. After the resulting male mice grew up till postnatal 8 week, CNO or vehicle PBS was intraperitoneally administered to the mice, and the pupil sizes of both eyes were measured in a dark room. B: Boxplot showing the pupil diameters from eyes at 20-50 mins after administration of PBS (pale yellow) and CNO (blue) in ICR mice whose cortices were in utero electroporated for hM3Dq at E12.5, E13.5, E14.5 and E15.5. The black line shows the difference of pupil diameters in individual eyes between PBS and CNO treatments. X marks the mean. One-tailed paired t-tests were performed between PBS and CNO treatments. C, D: Distribution of hM3Dq-positive areas and FOS-positive neurons in cortical layers of somatosensory areas on the in utero electroporated (EP) and unelectroporated (non-EP) sides. The same mice in B were analyzed at postmortem.

Article Snippet: pAAV-PTRE-tight -hM3Dq-mCherry was obtained from Dr William Wisden (Addgene plasmid # 66795) and the coding sequences for hM3Dq fused with mCherry were subcloned into a CAGGS expression plasmid .

Techniques: Activation Assay, Expressing, Plasmid Preparation, In Utero, One-tailed Test