AAV-hSyn1-GCaMP6s-P2A-nls-dTomato in Rat (AAV Retrograde)
Addgene #51084-AAVrg

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.

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.

Virus & Injection

Primary Virus
Promoter
Syn
Injected Titer
9.00E+12 GC/ml
Injected Volume
750 nl
Injection Rate
150 nl/min
Injection Site
Brain parenchyma
Brain Region
Midbrain: Tegmentum: Substantia nigra
Injection Site Comments
Targeting ventral tegmental area
View in Addgene Catalog

Subject

Species / Strain
Rat Sprague-Dawley from Envigo
Stock Number
Envigo rat stock no: 002
Age at Injection
6 weeks

Data Collection

Time After Injection
3 weeks
Detection Method
Two-photon calcium imaging
Expression Site
Direct pathway medium spiny neurons of the nucleus accumbens
Genetic Tool Outcome
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