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pAAV-CAG-GFP in Guinea pig (AAV Retrograde)

Addgene #37825-AAVrg

Data Submitted By
Nichole Beebe and Brett Schofield
Submission Date
August 14, 2019
Publication Date
August 14, 2019 (modified May 16, 2023)
Abstract
Not Provided
This is an image taken at 10X on a Zeiss AxioImager microscope. The tissue is paraformaldehyde-fixed, frozen-sectioned, then mounted onto a gelatin-coated slide and coverslipped with DPX mountant. This is direct EGFP fluorescence in pyramidal cells of layers V and VI of auditory cortex in a guinea pig. The fluorescence is the result of an injection of rgAAV-CAG-EGFP into the ipsilateral inferior colliculus of this animal.
Caption: This is an image taken at 10X on a Zeiss AxioImager microscope. The tissue is paraformaldehyde-fixed, frozen-sectioned, then mounted onto a gelatin-coated slide and coverslipped with DPX mountant. This is direct EGFP fluorescence in pyramidal cells of layers V and VI of auditory cortex in a guinea pig. The fluorescence is the result of an injection of rgAAV-CAG-EGFP into the ipsilateral inferior colliculus of this animal.
Image attribution: Taken by Brett Schofield.

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

Vector Used
pAAV-CAG-GFP
Addgene #37825-AAVrg

Virus & Injection

Virus

Virus Name
pAAV-CAG-GFP
Serotype
AAV Retrograde
Source
Addgene
Catalog Number
37825-AAVrg
Promoter
CAG
Cargo Type
Control
Injection Titer / Dose
5.00E+12 GC/ml
Injection Volume
two 0.05 μl injections (0.1 μl total)
Injection Rate
20 nL/min
Injector Material
Not Provided
Injection Site / Route
Inferior colliculus, NeuroNames ID : 476
Additional Details
The injection was made into the inferior colliculus in an attempt to label neurons in other brain regions (e. G. Contralateral inferior colliculus, lower brainstem auditory nuclei, auditory cortex) that make projections to the inferior colliculus.
View in Addgene Catalog

Other Details

Species
Other
Other Species
Guinea Pig
Strain or Cell Line
N/A
Source
Not provided
Other Information
The subject weighed 358 g (corresponding to an approximate age of 6-7 weeks).
Age at Injection
6-7 weeks
Time After Injection
5 weeks
Detection Method
Direct fluorescence
Assay & Results
Of all neuronal types that could potentially have expressed the virus, only pyramidal cells in the auditory cortex were detectable by observing direct fluorescence. This virus seems to work in a very pathway-specific manner (at least in guinea pigs). We expected that any neurons that make projections to the inferior colliculus might express EGFP in the tissue due to the general CAG promoter present in the virus, but only pyramidal cells in the auditory cortex were detectable by observing direct fluorescence
Target Cell Transduction
There were multiple brain areas where we should have seen fluorescent cells, and we only observed them in one area. The areas of input from these nuclei overlap in the injected nucleus, so the best explanation for this observation is cell-type specificity in either infection or expression.
Other Information
Very little toxicity or inflammation were present at the injection site