pAAV-hSyn-DIO-hM3D(Gq)-mCherry in Mouse (AAV8)
Addgene #44361-AAV8

Microscope: Olympus IX81; Objective: Olympus Plapon 2X; Filters: Excitation TRITC 575, Emission TRITC 624; Exposure 1000ms; Software: Metamorph Advanced. Image: Thick slice (250um) cut on a vibratome from a CaMKII-Cre mice injected with AAV8-hSyn-DIO-hM3Dq-mCherry. The image shows selective expression in dorsomedial prefrontal cortex in putative excitatory principal neurons. The signal comes from the mCherry fluorescence without any amplification.

Caption: Microscope: Olympus IX81; Objective: Olympus Plapon 2X; Filters: Excitation TRITC 575, Emission TRITC 624; Exposure 1000ms; Software: Metamorph Advanced. Image: Thick slice (250um) cut on a vibratome from a CaMKII-Cre mice injected with AAV8-hSyn-DIO-hM3Dq-mCherry. The image shows selective expression in dorsomedial prefrontal cortex in putative excitatory principal neurons. The signal comes from the mCherry fluorescence without any amplification.

Virus & Injection

Primary Virus
Promoter
Syn
Injected Titer
4.00E+12 GC/ml
Injected Volume
400nl
Injection Rate
0. 1nl/min
Injection Site
Brain parenchyma
Brain Region
Forebrain: Amygdala
Injection Site Comments
In addition to the amygdala we have also injected in prefrontal cortex (cingulate, prelimbic and infralimbic) and VTA. We used Cre lines for cellular selectivity
View in Addgene Catalog

Subject

Species / Strain
Mouse B6.Cg-Tg(Camk2a-cre)T29-1Stl/J, The Jackson Laboratory
Age at Injection
50-55 for females and 45-50 for males

Data Collection

Time After Injection
Between 3 and 5 weeks
Detection Method
Direct fluorescence, Patch-clamp electrophysiology in voltage clamp to detect changes in RMP and Rheobase
Expression Site
We observed expression in all brain regions tested and the cell type was given by the expression of Cre
Genetic Tool Outcome
CNO was used to activate hM3Dq and we observed changes in cellular excitability as expected (through slice electrophysiology recordings) as well as changes in behavior which depend on the brain region where the virus was infused
Tips & Comments
Fluorescence for this virus tends to be patchy being a fusion protein. Interested investigators should consider IRES or 2A peptide versions