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pAAV-hSyn-DIO-hM3D(Gq)-mCherry in Mouse (AAV Retrograde)

Addgene #44361-AAVrg

Data Submitted By
Michael Chiang and Sarah E. Ross
Submission Date
July 17, 2019
Publication Date
July 17, 2019 (modified May 16, 2023)
Abstract
Not Provided
blue - dapi red - mCherry Green - NeuN Microscope is a Olympus BX53 fluorescent microscope with UPlanSApo 4x primary antibody: Rabbit RFP at 1:1000; Mouse NeuN at 1:1000 secondary alexafluor 488; alexafluor 555 Dapi 1:10,000
Caption: blue - dapi red - mCherry Green - NeuN Microscope is a Olympus BX53 fluorescent microscope with UPlanSApo 4x primary antibody: Rabbit RFP at 1:1000; Mouse NeuN at 1:1000 secondary alexafluor 488; alexafluor 555 Dapi 1:10,000

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-AAVrg

Virus & Injection

Virus

Virus Name
pAAV-hSyn-DIO-hM3D(Gq)-mCherry
Serotype
AAV Retrograde
Source
Addgene
Catalog Number
44361-AAVrg
Promoter
Synapsin
Cargo Type
Chemogenetics
Injection Titer / Dose
1.00E+13 GC/ml
Injection Volume
300 nL
Injection Rate
100 nL/min
Injector Material
Not Provided
Injection Site / Route
Spinal cord, NeuroNames ID: 855
Additional Details
0.5 ul, 100 ul/min, KOR-cre neurons in spinal cord. Injections were in L4 level of spinal cord. Visualized retrogradely labeled cells from RVM in the KOR-cre mouse line generated within our lab.
View in Addgene Catalog

Other Details

Species
Mouse
Strain or Cell Line
KOR-cre
Source
N/A
Other Information
Mouse line generated within the Ross lab. Generation of a KOR-Cre knockin mouse strain to study cells involved in kappa opioid signaling. PMCID: PMC4747253 DOI: 10.1002/dvg.22910.
Age at Injection
16 weeks
Time After Injection
10 weeks
Detection Method
Immunohistochemistry
Assay & Results
Visualized retrogradely labeled cells from RVM in the KOR-cre mouse line generated within our lab.
Target Cell Transduction
The retro virus back-labeled to regions that are known to project to spinal cord, such as the RVM, so the assumption is that the tool has worked at least to some extent.
Other Information
We did not look for neuroinflammation or toxicity. However, it is difficult to rule out whether any cellular debris was due to this vs. tissue damage from the needle injection.
Tips & Comments
Would not change anything other than time for virus to express, which would need to be empirically tested (e.g. waiting 2, 4, 6, and 8 weeks).