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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
Caption: Figure 1: Interneurons in the mPFC innervated by BLA projections. Panel A shows the distribution of the tdTomato labeled neurons in the cingulate and prelimbic regions of the mouse mPFC. Panel B shows the morphology of the labeled neurons; small round soma with elaborate dendritic trees. Panel C shows a high magnification image of tdTomato labeled neuron, demonstrating the ability of the AAV-hDlx construct to completely fill up different compartments of the neuron.

Any further use of this image requires permission from the lab who submitted the data. This scientist responded to a specific request for user data and was offered compensation for time and input.

Vectors Used
pEEN.AAV.hSyn.Cre.WPRE.hGH, serotype AAV1
#105553
pAAV-hDlx-Flex-dTomato-Fishell_7
#83894

Virus & Injection

Virus 1

Virus Name
pEEN.AAV.hSyn.Cre.WPRE.hGH, serotype AAV1
Serotype
AAV1
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.
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Virus 2

Virus Name
pAAV-hDlx-Flex-dTomato-Fishell_7
Serotype
AAV9
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.
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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.