Interneurons in the medial prefrontal cortex innervated by amygdalar projections
Addgene #105553
- Data Submitted By
- Eugene Dimitrov
- Submission Date
- November 30, 2022
- Publication Date
- January 12, 2023 (modified May 16, 2023)
- DOI
- https://doi.org/10.57733/addgene.diro97
- Funding
- Intradepartmental, E. Dimitrov PI
- Abstract
- Medial prefrontal cortex (mPFC) receives strong input from the basolateral amygdala (BLA). We used post-synaptic cre transfer to visualize the interneuronal population in the mPFC innervated by BLA efferents.
- Citation
-
Interneurons in the medial prefrontal cortex innervated by amygdalar projections. Dimitrov E 2023. Addgene Report, https://doi.org/10.57733/addgene.diro97
This report is made available under the Creative Commons Attribution 4.0 International License.
- Vectors Used
-
pEEN.AAV.hSyn.Cre.WPRE.hGH, serotype AAV1
Addgene #105553 -
pAAV-hDlx-Flex-dTomato-Fishell_7
Addgene #83894
Virus & Injection
Virus 1
- Virus Name
- pEEN.AAV.hSyn.Cre.WPRE.hGH, serotype AAV1
- Serotype
- AAV1
- Source
- Addgene
- Catalog Number
- 105553
- Virus Lot Number
- v61518
- Promoter
- Synapsin
- Cargo Type
- Recombinase
- Injection Titer / Dose
- 1.80 x 10e13 vg/mL
- Injection Volume
- 100 nL
- Injection Rate
- 25 nL/min
- Injector Material
- Hamilton syringe
- Tip Diameter
- 210
- Injection Time
- 4-5 min
- Injection Site / Route
- Basolateral Amygdala, NeuroNames ID: 246
- Additional Details
- 100 nL of undiluted viral prep was injected with a Hamilton air-tight syringe (33 Ga) attached to microinjection pump at a rate 25 microliters per minute. The stereotaxic coordinates for the injection were -1.8 mm, -3.0 mm and - 4.5 mm to bregma according to the Mouse Brain Atlas.
Virus 2
- Virus Name
- pAAV-hDlx-Flex-dTomato-Fishell_7
- Serotype
- AAV9
- Source
- Addgene
- Catalog Number
- 83894
- Virus Lot Number
- v94956
- Promoter
- hDLX
- Cargo Type
- Control
- Injection Titer / Dose
- 2.9 x 10e13 vg/mL
- Injection Volume
- 200 nL
- Injection Rate
- 25 nL/min
- Injector Material
- Hamilton syringe
- Tip Diameter
- 210
- Injection Time
- 5 min
- Injection Site / Route
- mPFC, prelimbic area, NeuroNames ID: 1078
- Additional Details
- 200 nL of undiluted viral prep was injected with 33 Ga needle and a Hamilton syringe attached to microinjection pump at rate 25 nL per minute. The stereotaxic coordinates for the injection into mPFC were +1.7 mm, -0.4 mm and - 2.0 mm to bregma according to the Mouse Brain Atlas. This virus was injected during the same surgery as the AAV1-cre virus.
Other Details
- Species
- Mouse
- Strain or Cell Line
- CD1
- Source
- Charles River
- Stock Number
- 1GS # 022
- Age at Injection
- 2 months
- Sex
- Female
- Time After Injection
- 21 Days
- Detection Method
- Native Fluorescence
- Assay & Results
- Mouse brains were fixed in 4% PFA, the mPFC region was cut into 40-micron thick sections, which were mounted with aqueous medium. Leica DM 5500 light microscope equipped with Hamamatsu CCD camera was used for imaging. The cre-dependent AAV9-hDlx-Flex-tdTomato-Fishell_7 was amply expressed by the neurons of the cingulate and prelimbic subdivisions of the mPFC. The cre was delivered via post-synaptic transfer from efferents originating in the BLA. The result indicates that the mPFC interneurons are directly targeted by BLA projections, therefore the activity of cortical interneurons is very likely under direct BLA control. Furthermore, the result shows that a floxed AAV with Dlx promoter can be activated by post-synaptic cre transfer, which provides a new approach for anatomical and functional studies of the brain interneuronal populations.
- Other Information
- This report is made available under the Creative Commons Attribution 4.0 International License.