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

Pontine nucleus in brainstem of mouse. The image was acquired by All-in-one fluorescent microscope (BZ-X800, Keyence). Green: GCaMP6s axons from cortex, blue: native EBFP, red: fluorescent Nissle (NeuroTrace)

Caption: Pontine nucleus in brainstem of mouse. The image was acquired by All-in-one fluorescent microscope (BZ-X800, Keyence). Green: GCaMP6s axons from cortex, blue: native EBFP, red: fluorescent Nissle (NeuroTrace)

Cortical neurons projecting to the pontine nuclei. The image was acquired by an all-in-one fluorescent microscope (BZ-X800, Keyence). Green: native GCaMP6s fluorescence.  Retrogradely labelled pontine-projecting cortical neurons.

Caption: Cortical neurons projecting to the pontine nuclei. The image was acquired by an all-in-one fluorescent microscope (BZ-X800, Keyence). Green: native GCaMP6s fluorescence. Retrogradely labelled pontine-projecting cortical neurons.

Image attribution: Retrogradely labelled pontine-projecting cortical neurons.

Viruses & Injection

Primary Virus
Promoter
Syn
Injected Titer
6.90E+12 GC/ml
Injected Volume
300 nl
Injection Rate
30 nl/min
Injection Site
Brain parenchyma
Brain Region
Hindbrain: Pons
Injection Site Comments
To label projection neurons to the target region
View in Addgene Catalog
Secondary Virus
AAV-hSyn-FLEX-GCaMP6s
Promoter
Syn
Injected Titer
1.00E+13 GC/ml
Injected Volume
200 nl
Injection Rate
20 nl/min
Injection Site
Brain parenchyma
Brain Region
Forebrain: Cerebral cortex (neocortex)
Injection Site Comments
To label projection neurons. Simultaneously injected

Subject

Species / Strain
Mouse C57/BL6J
Age at Injection
8-12 weeks

Data Collection

Time After Injection
2 weeks
Detection Method
Direct fluorescence
Expression Site
Injection site (FLEX virus) specific
Other Information
The tropism of AAV serotypes, especially about AAVrg
Tips & Comments
If you prevent the back-flow of viral vector, I recommend the injection speed is set as slower as possible. In our lab. , serotype 1 and rg (for antero- and retrograde labelling) generally were used. But I wonder rg virus is strongest expression-level at injection site (that is, anterograde-labelling manner), so I worry about the cytotoxicity. I think the lower titer (~1E11 gc/mL) might be suitable