pAAV-hSyn-DIO-mCherry in Mouse (AAV2)
Addgene #50459-AAV2

Olympus BX53 fluorescent microscope with UPlanSApo  20x, blue is DAPI, red is rabbit-RFP enhanced mCherry with AlexaFluor 555. Primary antibody: rockland rabbit anti-RFP 1:1000
Secondary antibody: alexafluor 555

Caption: Olympus BX53 fluorescent microscope with UPlanSApo 20x, blue is DAPI, red is rabbit-RFP enhanced mCherry with AlexaFluor 555. Primary antibody: rockland rabbit anti-RFP 1:1000 Secondary antibody: alexafluor 555

Virus & Injection

Primary Virus
Promoter
Syn
Injected Titer
1.00E+13 GC/ml
Injected Volume
500 nl
Injection Rate
100 nl/min
Injection Site
Brain parenchyma
Brain Region
Midbrain - parabrachial nucleus
Injection Site Comments
Gad2-cre, inhibitory population
View in Addgene Catalog

Subject

Species / Strain
Mouse Gad2-IRES-cre on C57BL/6J background
Stock Number
10802 from Jackson Labs
Age at Injection
8 weeks

Data Collection

Time After Injection
4 weeks
Detection Method
Immunohistochemistry
Expression Site
Within lateral parabrachial nucleus
Genetic Tool Outcome
The fluorophore was expressed within a subpopulation of lateral parabrachial nucleus which need to be further characterized using FISH (experiments ongoing)
Other Information
AAV2 does not seem to cause neuronal blebbing as depicted here: doi: 10. 3389/fncir. 2013. 00008. AAV9 seems to be neurotoxic
Tips & Comments
EF1a, CMV, CAG promoters are very strong but seem to result in fewer number of neurons infected compared to AAV under hsyn promoter