Skip to main content
Return to AAV Results

Testing AAVrg-hSyn-mCherry in the Spinal Cord of Rhesus Macaques

Addgene #114472-AAVrg

Data Submitted By
Adriana Galvan and Johnson Agniswamy
Submission Date
August 04, 2023
Publication Date
August 24, 2023 (modified August 24, 2023)
DOI
https://doi.org/10.57733/addgene.s6f8sp
Funding
Aligning Science Across Parkinson's ASAP-020572 through the Michael J. Fox Foundation for Parkinson’s Research (MJFF) ( Wichmann, PI, Galvan Co-I); P50NS123103 (NIH/NINDS) (Wichmann, PI, Galvan Co-I); P51-OD011132 (Emory Primate Center base grant); ASAP-020572; P50NS123103; P51-OD011132
Abstract
Two rhesus macaque animals were tested for AAVrg-hSyn-mCherry in the spinal cord. We observed abundant mCherry-positive neurons in expected targets (i.e., motor cortical regions known to project to the spinal cord).
Citation
Testing AAVrg-hSyn-mCherry in the Spinal Cord of Rhesus Macaques. Galvan A, Agniswamy J 2023. Addgene Report, https://doi.org/10.57733/addgene.s6f8sp
Injection of AAVrg-hSyn-mCherry in the spinal cord of rhesus monkey. A) mCherry labeling in the left spinal cord. The putative injection track is indicated with an arrow. Note that the virus solution resulted in profuse labeling of neuropil (putative axonal terminals). B) mCherry-positive neurons in supraspinal regions (opposite hemisphere to the injection site). Abundant mCherry-positive cells were observed in the primary motor cortex (M1) and in the supplementary motor region (SMA). Note the labeled axons of the internal capsule (i.c.). C) Higher magnification of the area indicated in a rectangle in (B). The mCherry-positive cells are located in deep layers, as expected for cortico-spinal projection neurons.
Caption: Injection of AAVrg-hSyn-mCherry in the spinal cord of rhesus monkey. A) mCherry labeling in the left spinal cord. The putative injection track is indicated with an arrow. Note that the virus solution resulted in profuse labeling of neuropil (putative axonal terminals). B) mCherry-positive neurons in supraspinal regions (opposite hemisphere to the injection site). Abundant mCherry-positive cells were observed in the primary motor cortex (M1) and in the supplementary motor region (SMA). Note the labeled axons of the internal capsule (i.c.). C) Higher magnification of the area indicated in a rectangle in (B). The mCherry-positive cells are located in deep layers, as expected for cortico-spinal projection neurons.

This report is made available under the Creative Commons Attribution 4.0 International License.

Vector Used
pAAV-hSyn-mCherry
Addgene #114472-AAVrg

Virus & Injection

Virus

Virus Name
pAAV-hSyn-mCherry
Serotype
AAVrg
Source
Addgene
Catalog Number
114472-AAVrg
Virus Lot Number
v111416
Promoter
Synapsin
Cargo Type
Control
Injection Titer / Dose
2.00 E+13 vg/ml
Injection Volume
2 μl per track, 6 tracks (total of 12 µl)
Injection Rate
2.5 µl/min
Injector Material
Custom-made injection cannula
Tip Diameter
30
Injection Time
The needle was withdrawn immediately after the spinal cord injection.
Injection Site / Route
Spinal Cord, C6-T1. Neuronames ID: 855
Additional Details
Six tracks of 2 µl of the viral vector were injected, adding to a total of 12 μL with an infusion rate of 2.5 μL/min using a custom-made injection device connected to a Hamilton microsyringe (Reno, NE) coupled to a microinfusion pump. The needle was withdrawn immediately after the injection.
View in Addgene Catalog

Other Details

Species
Non-Human Primate
Strain or Cell Line
Rhesus Macaque
Source
Emory National Primate Research Center
Other Information
1 female, 1 male
Age at Injection
4 years, 10 months; 3 years 10 months
Sex
Mixed
Time After Injection
6 weeks
Detection Method
IHC
Assay & Results
Two animals were injected in the spinal cord. We used antibodies against mCherry to examine the FP expression. The brain sections were pretreated (10% normal goat serum, 1% bovine serum albumin, and 0.3% Triton X-100) and then incubated in the antibody solution for 24 h. The sections were then incubated in the biotinylated antibody for 90 min. Afterward, the sections were placed in 0.025% 3-3’-diaminobenzidine tetrachloride, 0.01 M Imidazole, and 0.006% H2O2 for 10 min. The sections were mounted on slides, covered, and digitized with an Aperio Scanscope CS system.
Target Cell Transduction
We observed abundant mCherry-positive neurons in expected targets (i.e., motor cortical regions known to project to the spinal cord)