pAAV-Ef1a-DIO hChR2(E123A)-EYFP in Rat (AAV9)
Addgene #35507-AAV9

Prelimbic cortex neurons projecting to thalamic/habenular areas are shown expressing Ef1a-DIO-hChR2(E123A)-EYFP. Immuno-amplification of EYFP was performed (Goat-anti GFP, AlexaFluor488) to allow visualization of virally delivered protein. Dorsal to the EYFP+ neurons is the track from an implanted optic fiber used for laser light delivery.

Caption: Prelimbic cortex neurons projecting to thalamic/habenular areas are shown expressing Ef1a-DIO-hChR2(E123A)-EYFP. Immuno-amplification of EYFP was performed (Goat-anti GFP, AlexaFluor488) to allow visualization of virally delivered protein. Dorsal to the EYFP+ neurons is the track from an implanted optic fiber used for laser light delivery.

Viruses & Injection

Primary Virus
Promoter
EF1a
Injected Titer
2.50E+13 GC/ml
Injected Volume
400 nl
Injection Rate
1 nl/sec
Injection Site
Brain parenchyma
Brain Region
Forebrain: Cerebral cortex (neocortex)
Injection Site Comments
Prelimbic Cortex neurons projecting to medial dorsal thalamic/habenular area
View in Addgene Catalog
Secondary Virus
pENN.AAV.hSyn.Cre.WPRE.hGH (AAV Retrograde)
Promoter
Syn
Injected Titer
1.20E+13 GC/ml
Injected Volume
200 nl
Injection Rate
1 nl/sec
Injection Site
Brain parenchyma
Brain Region
Forebrain: Epithalamus
View in Addgene Catalog

Subject

Species / Strain
Rat Long-Evans
Age at Injection
~3 months old

Data Collection

Time After Injection
8 weeks
Detection Method
Immunohistochemistry
Expression Site
Prelimbic cortex, corticothalamic projection neurons
Genetic Tool Outcome
Pilot testing virus expression and optic fiber placement. No light was used in these experiments to activate the opsin
Other Information
The only inflammation/glial scarring that was observed was a result of optic fiber implantation above the region of viral expression. Dexamethasone treatment prior to and for ~3 days after surgery may help to reduce inflammation around optic fiber site