pAAV-Ef1a-DIO hChR2(E123A)-EYFP in Rat (AAV9)
Addgene #35507-AAV9
-EYFP+Optic FIber_PrelimbicCortex-Thalamic Projections.jpg)
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
- 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
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