AAV pmSyn1-EBFP-Cre in Mouse (AAV Retrograde)
Addgene #51507-AAVrg

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

Virus & Injection

Primary Virus
Promoter
Syn
Injected Titer
5.00E+12 GC/ml
Injected Volume
two 400 nl injections
Injection Rate
75 nl/min
Injection Site
Brain parenchyma
Brain Region
Hindbrain: Pons
Injection Site Comments
Lateral Parabrachial Nucleus was targeted in an effort to retrogradely label spino-parabrachial projection neurons in lamina I of the spinal cord
View in Addgene Catalog

Subject

Species / Strain
Mouse B6;129-Gt(ROSA)26Sortm5(CAG-Sun1/sfGFP)Nat/J
Stock Number
21039
Age at Injection
62 days

Data Collection

Time After Injection
3 weeks
Detection Method
Direct fluorescence
Expression Site
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
Genetic Tool Outcome
The mouse line expresses a Sun1-GFP fusion protein in the presence of cre recombinase; 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