Labeling Trigeminal Ganglion Afferents with PNS-Trophic AAV
Addgene #59462
- Data Submitted By
- JJ Kim and Dan O'Connor
- Submission Date
- November 29, 2022
- Publication Date
- January 12, 2023 (modified May 16, 2023)
- DOI
- https://doi.org/10.57733/addgene.abmgfd
- Funding
- NIH/NINDS R01NS084818
- Abstract
- We tested the labelling of trigeminal ganglion afferents in maxillary division (V2) with stereotactic injection of AAV.PHP.S-CAG-tdTomato. The PNS-trophic AAV labelled the sensory ganglion robustly with clear labelling of cell bodies and axons. Expression was localized to V1/V2 region of the ganglion.
- Citation
-
Labeling Trigeminal Ganglion Afferents with PNS-Trophic AAV. Kim J, O'Connor D 2023. Addgene Report, https://doi.org/10.57733/addgene.abmgfd
This report is made available under the Creative Commons Attribution 4.0 International License.
- Vector Used
-
pAAV-CAG-tdTomato
Addgene #59462
Virus & Injection
Virus
- Virus Name
- pAAV-CAG-tdTomato
- Serotype
- PHP.S
- Source
- Addgene
- Catalog Number
- 59462
- Virus Lot Number
- v47316
- Promoter
- CAG
- Cargo Type
- Control
- Injection Titer / Dose
- 2.1 x 10e13
- Injection Volume
- 300 nL
- Injection Rate
- 50nL/min
- Injector Material
- pulled glass pipette
- Injection Time
- 5 min
- Injection Site / Route
- Trigeminal ganglion, NeuroNames ID: 1402
- Additional Details
- Undiluted viral titer (1.1E13 GC/mL) was injected at the following stereotactic coordinates (bregma) based on tungsten electrode recordings of trigeminal ganglion V2: AP 0, ML 0.9, DV 6.3-6.5. The pipette was a pulled glass capillary micropipette (Wiretrol II, Drummond Scientific Company) with bevel of 30 deg and outer diameter of 50-70 um. Pipette was loaded by tip filling and advanced slowly to target. Injection occured at 50 nL/min rate with hydraulic micromanipulator (Narishige, MO-10) and at 2 depths DV 6.3 and 6.5 mm. The pipettte was kept in place for 5-10 minutes prior to retraction.
Other Details
- Species
- Mouse
- Strain or Cell Line
- C57/BL6
- Source
- Jackson Labs
- Stock Number
- 000664
- Age at Injection
- 8-12 Weeks
- Sex
- Male
- Time After Injection
- 21 Days
- Detection Method
- Native Fluorescence
- Assay & Results
- Native fluorescence was assayed with lightsheet imaging of cleared whole trigeminal ganglion. tdTomato expression with local stereotactic injection of AAV PHP.S was robust and centered on the maxillary division of trigeminal ganglion with clear labelling of axonal tracts. This result was consistent in N=2 animals tested with no expression in the uninjected contralateral trigeminal ganglion.
- Other Information
- This report is made available under the Creative Commons Attribution 4.0 International License.