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

Addgene #44361-AAV8

Data Submitted By
Ezequiel Marron and Kevin Wickman
Submission Date
July 17, 2019
Publication Date
July 17, 2019 (modified May 16, 2023)
Abstract
Not Provided
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.

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

Vector Used
pAAV-hSyn-DIO-hM3D(Gq)-mCherry
Addgene #44361-AAV8

Virus & Injection

Virus

Virus Name
pAAV-hSyn-DIO-hM3D(Gq)-mCherry
Serotype
AAV8
Source
Addgene
Catalog Number
44361-AAV8
Promoter
Synapsin
Cargo Type
Chemogenetics
Injection Titer / Dose
4.00E+12 GC/mL
Injection Volume
400 nL
Injection Rate
100 nL/min
Injector Material
Not Provided
Injection Site / Route
Amygdala, NeuroNames ID: 237
View in Addgene Catalog

Other Details

Species
Mouse
Strain or Cell Line
B6.Cg-Tg(Camk2a-cre)T29-1Stl/J,
Source
Jackson Labs
Age at Injection
50-55 for females and 45-50 for males
Sex
Mixed
Time After Injection
Between 3 and 5 weeks
Detection Method
Direct fluorescence
Assay & Results
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. We observed expression in all brain regions tested and the cell type was given by the expression of Cre.
Functional Testing
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. Patch-clamp electrophysiology in voltage clamp to detect changes in RMP and Rheobase.
Tips & Comments
Fluorescence for this virus tends to be patchy being a fusion protein. Interested investigators should consider IRES or 2A peptide versions.