aav hsynflex mgfp 2a synaptophysin mruby  (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 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

Journal: bioRxiv

Article Title: Connectivity and phenotype of vasopressin 1a receptor cells in the lateral septum

doi: 10.1101/2025.10.26.683946

Figure Lengend Snippet: Anterograde tracing of LS V1aR cells. A. Representative image of unilateral injection of Synaptophysin AAV into the LS (10x, Scale bar = 50μm). B. Example of injection spread in the LS. C. Example images of fiber densities for each semiqualitative ranking: +, very few fibers; ++, few dispersed fibers; +++, part of region densely covered by fibers; ++++, most of region densely covered by fibers (40x, Scale bars = 50μm). D. Example images of puncta densities for each semiqualitiative ranking: +, very few to no puncta; ++, few dispersed puncta; +++ some puncta; ++++ dense puncta across region (40x, Scale bars = 50μm). E. Examples of fibers and puncta in regions of interest in one female brain: medial septum, horizontal diagonal band, lateral preoptic area, anterior hypothalamic area, ventral pallidum, penducular lateral hypothalamus, and supramammillary nucleus (40x, Scale bars = 50μm). F . Representative diagram of fiber density and injection spread throughout one female brain.

Article Snippet: We identified the neural targets of LS Avpr1a cells by placing unilateral stereotaxic injections of an anterograde adeno-associated virus (AAV) tracer (hSyn-Flex-mGFP-2A-Synaptophysin-mRuby; Addgene #71760) into the LS of Avpr1a Cre+ (N9 generation) males (n=1/sex) and females (n=1/sex) as well as Avpr1a Cre- subjects (n=2/sex) as controls.

Techniques: Anterograde Tracing, Injection