pAAV-hSyn-DIO-hM3D(Gq)-mCherry in Mouse (AAV Retrograde)
Addgene #44361-AAVrg

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

Virus & Injection

Primary Virus
Promoter
Syn
Injected Titer
1.00E+13 GC/ml
Injected Volume
300 nl
Injection Rate
100 nl/min
Injection Site
Brain parenchyma
Brain Region
Spinal cord
Injection Site Comments
0. 5 ul, 100 ul/min, KOR-cre neurons in spinal cord
View in Addgene Catalog

Subject

Species / Strain
Mouse KOR-cre 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

Data Collection

Time After Injection
10 weeks
Detection Method
Immunohistochemistry
Expression Site
Injections were in L4 level of spinal cord, visualized retrogradely labeled cells from RVM in the KOR-cre mouse line generated within our lab
Genetic Tool Outcome
The retro virus backlabeled 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