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pAAV.Syn.NES-jRGECO1a.WPRE.SV40 in Rat (AAV Retrograde)

Addgene #100854-AAVrg

Data Submitted By
Elizabeth Crummy and Susan Ferguson
Submission Date
June 21, 2019
Publication Date
June 21, 2019 (modified May 16, 2023)
Abstract
Not Provided
Caption: 488 channel for GCaMP expression imaging on a Zeiss LSM 710 confocal 10x.
Caption: Imaged in 568 channel on a Zeiss LSM 710 confocal at 10x.

Any further use of this image requires permission from the lab who submitted the data.

Vectors Used
pAAV.Syn.NES-jRGECO1a.WPRE.SV40
Addgene #100854-AAVrg
AAV-hSyn1-GCaMP6s-P2A-nls-dTomato
Addgene #51084-AAVrg

Virus & Injection

Virus 1

Virus Name
pAAV.Syn.NES-jRGECO1a.WPRE.SV40
Serotype
AAV Retrograde
Source
Addgene
Catalog Number
100854-AAVrg
Promoter
Synapsin
Cargo Type
Biosensor
Injection Titer / Dose
1.00E+13 GC/ml
Injection Volume
1000 nL
Injection Rate
200 nL/min
Injector Material
Not Provided
Injection Site / Route
Nucleus accumbens, NeuroNames ID: 277
View in Addgene Catalog

Virus 2

Virus Name
AAV-hSyn1-GCaMP6s-P2A-nls-dTomato
Serotype
AAV Retrograde
Source
Addgene
Catalog Number
51084-AAVrg
Promoter
Synapsin
Injection Titer / Dose
9.00E+12 GC/ml
Injection Volume
1000 nl
Injection Rate
200 nl/min
Injector Material
Not Provided
Injection Site / Route
Pons, NeuroNames ID: 547
Additional Details
Pyramidal tract (PT) cortical projection neurons projecting to the pons. Injected at same session as jRGECO.
View in Addgene Catalog

Other Details

Species
Rat
Strain or Cell Line
Sprague Dawley
Source
Not provided
Age at Injection
~2-3 months old
Time After Injection
3 weeks
Detection Method
Immunohistochemistry, fiber photometry recordings
Assay & Results
GCaMP PT expression, jRGECO IT expression.
Target Cell Transduction
Targeting intratelencephalic (IT) cortical neurons projecting to the nucleus accumbens. GCaMP PT expression, jRGECO IT expression.
Functional Testing
We have had issues with obtaining functional recordings during fiber photometry. Have had issues with jRGECO not expressing in some cohorts.
Tips & Comments
Maybe try to dilute the virus (particuarly GCaMP) and inject a lower volume.