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AAV pmSyn1-EBFP-Cre in Mouse (AAV Retrograde)

Addgene #51507-AAVrg

Data Submitted By
Elizabeth Serafin and Mark Baccei
Submission Date
June 19, 2019
Publication Date
June 19, 2019 (modified May 16, 2023)
Abstract
Not Provided
This image shows a 20um fixed-frozen horizontal section obtained through lamina I of the spinal cord, contralateral to site of brain injection. Fluorescence in FITC channel indicates expression of nuclear Sun1-GFP fusion protein in the presence of virally expressed cre recombinase. White arrows indicate GFP+ nuclei; the location and distribution are consistent with that of lamina I spino-PB projection neurons. Image taken on Olympus BX63 upright fluorescent microscope using 10X objective.
Caption: This image shows a 20um fixed-frozen horizontal section obtained through lamina I of the spinal cord, contralateral to site of brain injection. Fluorescence in FITC channel indicates expression of nuclear Sun1-GFP fusion protein in the presence of virally expressed cre recombinase. White arrows indicate GFP+ nuclei; the location and distribution are consistent with that of lamina I spino-PB projection neurons. Image taken on Olympus BX63 upright fluorescent microscope using 10X objective.
Image attribution: Elizabeth Serafin

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

Vector Used
AAV pmSyn1-EBFP-Cre
Addgene #51507-AAVrg

Virus & Injection

Virus

Virus Name
AAV pmSyn1-EBFP-Cre
Serotype
AAV Retrograde
Source
Addgene
Catalog Number
51507-AAVrg
Promoter
Synapsin
Cargo Type
Recombinase
Injection Titer / Dose
5.00E+12 GC/mL
Injection Volume
two 400 nL injections
Injection Rate
75 nL/min
Injector Material
Not Provided
Injection Site / Route
Parabrachial nucleus, NeuroNames ID : 1927
Additional Details
Lateral Parabrachial Nucleus was targeted in an effort to retrogradely label spino-parabrachial projection neurons in lamina I of the spinal cord; Hindbrain: Pons.
View in Addgene Catalog

Other Details

Species
Mouse
Strain or Cell Line
B6;129-Gt(ROSA)26Sortm5(CAG-Sun1/sfGFP)Nat/J
Source
Jackson Labs
Stock Number
21039
Age at Injection
62 days
Time After Injection
3 weeks
Detection Method
Direct fluorescence
Assay & Results
Robust and widespread expression at injection site in parabrachial nucleus. A moderate number of target cells (projection neurons in lamina I of the spinal cord) also expressed genetic cargo.
Target Cell Transduction
Nuclear GFP fluorescence was visible as expected following injection of the AAVrg-driven cre. So yes, the tool operated as expected, although the number of retrogradely transduced neurons was somewhat lower than expected.
Other Information
I also checked 2 weeks post-injection and found a similar pattern of expression, though the number of retrogradely transduced neurons was less than the three week timepoint
Tips & Comments
This is one of the better retrograde viruses I have tried, though the number of transduced target cells was still lower than desired. To improve results with this virus I would probably try to increase injection volume or use multiple injection sites.