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AAV-hSyn1-GCaMP6s-P2A-nls-dTomato in Rat (AAV Retrograde)

Addgene #51084-AAVrg

Data Submitted By
Tim O'neal and Susan Ferguson
Submission Date
July 05, 2019
Publication Date
July 05, 2019 (modified May 16, 2023)
Abstract
Not Provided
Caption: Two-photon calcium imaging, imaged at 20x, 470nm excitation. The referenced image shows expression of the GCaMP in the green channel. The sensor expressed normally, but there was no calcium-dependent changes in fluorescence that we were able to detect.

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

Vector Used
AAV-hSyn1-GCaMP6s-P2A-nls-dTomato
Addgene #51084-AAVrg

Virus & Injection

Virus

Virus Name
AAV-hSyn1-GCaMP6s-P2A-nls-dTomato
Serotype
AAV Retrograde
Source
Addgene
Catalog Number
51084-AAVrg
Promoter
Synapsin
Cargo Type
Biosensor
Injection Titer / Dose
9.00E+12 GC/ml
Injection Volume
750 nl
Injection Rate
150 nl/min
Injector Material
Not Provided
Injection Site / Route
Ventral tegmental area, NeuroNames ID: 521
Additional Details
Targeting ventral tegmental area. Direct pathway medium spiny neurons of the nucleus accumbens
View in Addgene Catalog

Other Details

Species
Rat
Strain or Cell Line
Sprague-Dawley
Source
Envigo
Stock Number
002
Age at Injection
6 weeks
Time After Injection
3 weeks
Detection Method
Two-photon calcium imaging
Assay & Results
The sensor expressed normally, but there was no calcium-dependent changes in fluorescence that we were able to detect.
Functional Testing
No - we tried virus undiluted and at three separate dilutions (1:5, 1:10, 1:15 in PBS), and though cells expressed GCaMP, there was never any detectable signal in vivo or ex vivo.
Other Information
No noticeable problems with cell morphology, but the virus did not work in our hands
Tips & Comments
We moved on to the AAVrg-GCaMP7s (104487-AAVrg), which has worked fantastically under the same conditions.