pAAV-EF1a-double floxed-hChR2(H134R)-mCherry-WPRE-HGHpA in Mouse (AAV9)
Addgene #20297-AAV9

Olympus BX53 fluorescent microscope with UPlanSApo 10X objective  - red indicates cell bodies expressing transgene  - blue indicates dapi  antibody was enhanced with a rockland rabbit anti-RFP primary antibody and alexafluor 555

Caption: Olympus BX53 fluorescent microscope with UPlanSApo 10X objective - red indicates cell bodies expressing transgene - blue indicates dapi antibody was enhanced with a rockland rabbit anti-RFP primary antibody and alexafluor 555

Virus & Injection

Primary Virus
Promoter
EF1a
Injected Titer
2.88E+13 GC/ml
Injected Volume
500 nl
Injection Rate
100 nl/min
Injection Site
Brain parenchyma
Brain Region
Midbrain - parabrachial nucleus
Injection Site Comments
Crh-cre population expresses the corticotropin releasing hormone intended to target lateral parabrachial nucleus
View in Addgene Catalog

Subject

Species / Strain
Mouse B6(Cg)-Crhtm1(cre)Zjh/J stock ID: 012704
Stock Number
12704
Age at Injection
8 weeks

Data Collection

Time After Injection
4 weeks
Detection Method
Immunohistochemistry
Expression Site
Although the injection targeted lateral parabrachial nucleus, we observed expression in the adjacent region Kolliker Fuse (KF). We have not confirmed whether these neurons are excitatory or inhibitory
Genetic Tool Outcome
We observed expression of virus in these the Crh-cre line. However, we have not confirmed whether these are indeed expressing Crh
Other Information
No obvious excitotoxicity was observed - the cell bodies did not exhibit any blebbing
Tips & Comments
Perhaps reduction in virus volume (but the expression appeared localized)