pAAV-EF1a-double floxed-hChR2(H134R)-EYFP-WPRE-HGHpA in Mouse (AAV Retrograde)
Addgene #20298-AAVrg

Left panel: Immunohistochemical labeling of Cre-dependent GFP expression via retrograde transgene delivery. Channel: 488 nm.  Right panel: Immunohistochemical labeling of GnRH. Channel: 647 nm.

Caption: Left panel: Immunohistochemical labeling of Cre-dependent GFP expression via retrograde transgene delivery. Channel: 488 nm. Right panel: Immunohistochemical labeling of GnRH. Channel: 647 nm.

Image attribution: Gerry Coughlin

Virus & Injection

Primary Virus
Promoter
EF1a
Injected Titer
1.00E+13 GC/ml
Injected Volume
200 nl bilaterally
Injection Rate
0. 02 μl/min
Injection Site
Brain parenchyma
Brain Region
Forebrain: Hypothalamus
Injection Site Comments
GnRH neurons in the median eminence
View in Addgene Catalog

Subject

Species / Strain
Mouse Tg(Gnrh1-cre)1Dlc/J
Stock Number
JAX: 021207
Age at Injection
8 weeks

Data Collection

Time After Injection
3 weeks
Detection Method
Immunohistochemistry, Direct fluorescence
Expression Site
GnRH neurons in the median preoptic nucleus (MnPO)
Genetic Tool Outcome
Yes. We saw retrograde expression in expected regions after confirming endogenous fluorescence with immunohistochemistry. Optogenetic manipulations were not performed due to the sparseness of the labeled population
Other Information
We did not see any toxicity