I am looking for a way to visualize activated GABAergic neurons after administration of a drug. I am in search of a promising protocol.
We published a method for c-fos/GAD double-immunolabelling a while back: Illig KR and Haberly LB (2003) J. Comp. Neurol. 457:361-373.
We did Fos and NeuN double labeling in recent JNC publication and you can check the protocol. http://www.ncbi.nlm.nih.gov/pubmed/23895375
The specific solution will depend very much on the particular primary antibodies that you have, so any published protocol that uses any other antibodies will have to be modified.
Check with Andy Fischer in Neurobiology at Ohio State. He is a master at this sort of immunolabeling.
We published an article about activation of GABA neurons in a mesopontine region in response to various drugs administration (antidepressants, psychostimulants). We used FosB/DeltaFosB staining to see activated GABA neurons which is allo a good way for what you want to do. Article: http://www.ncbi.nlm.nih.gov/pubmed/21087442
Good luck !
I'd also add that cFos (and related transcription factors, particularly Fra-1 and -2) are not the only way to go. Egr-1 is another candidate; see Zhong et al., Invest Ophthal Vis Sci 2004;45:2065–2074 -- visual induction of Egr-1 in GABA neurons of macaque retina.
Hi Sara, I would recommend combining in situ hybridisation for GAD67 with IHC detection of c-fos. These two studies from our lab have recently been published using the technique http://link.springer.com/article/10.1007/s00429-013-0642-3 and http://onlinelibrary.wiley.com/doi/10.1002/cne.23310/full and if you're keen for a good ISH probe sequence for GAD67, check out http://onlinelibrary.wiley.com/doi/10.1002/cne.23175/full
I can send you the papers if you can't access the full text. All the best!
Which brain region are you targeting? If it is big enough to perform western blot I'd recommend you use this procedure.
Wow, thank you guys very much! These are all very helpful papers and links. Dayan- I intend on looking at the mPFC. I have considered western blots but I think I prefer IHC/IFC.
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