Our lab is interested in doing single-unit recordings in area CA2 of the hippocampus. We have a lot of experience with single-unit recording, but not in area CA2. Does anyone have experience trying to identify this area through histology? We are interested in both dorsal and ventral hippocampus.
You can use a PCP4 antibody to label CA2 after your recordings.
Article RGS14 Restricts Plasticity in Hippocampal CA2 by Limiting Po...
Thanks for your response Paula. I've seen this article before, but I noticed they used transgenic mice expressing Amigo2-EGFP. Do you know if PCP-4 needs to be used with these transgenic mice in order to work? Our lab typically uses rats, so I'm unfamiliar with this area.
Yes you can use PCP4 in rats, and also you can use RGS14 AB both will specifically label the CA2 region. For the calcium experiments we used organotypic slices of regular WT and Rgs14 KO animals (not the amigo2 line) and patch cells with a solution containing alexa, then fixed the slices, and do a post immuno's to verify that the cell was really in CA2. Good luck!
Perfect! I didn't even notice I was speaking to one of the authors! Really interesting paper and great work. Thanks for the help!
For anyone using this thread for their own purposes... Our lab used anti-rabbit PCP4 antibody (1:250 dilution) and donkey anti-rabbit Alexa Fluor 488 (1:200 dilution) and achieved some nice stains.
Hi,
I doing my PhD in a lab of animal behavior. We work with mouse and study social behaviors related to autism. We are interested in identify CA2 area in the hippocampus to, possibly, observed differences between treated and no treated mice. Literature showed as in situ hybridization, but it is a complex technic. Do you think that your methods can helps as to identified this area? And, moreover, do you cut your rat brains with a cryostat?
Thanks
Hi Judit Biosca Brull ,
Sure, happy to help. We were able to identify CA2 pretty well with our method and I think this method should work for your purposes. See my answer on this thread above from September 27th, 2018. Note that since then, we have changed the concentration of the antibodies to 1:500 for the PCP4 antibody and 1:1000 for the Alexa Fluor 488. We used a vibratome to section our brains, although I think a cryostat would be fine as well. I'm happy to share our protocol with you if you would like.
Best,
Ryan
Thanks for your help! We are looking for different techniques to see which is the most optimal and which we can perform in our laboratory. If you don’t mind I would appreciate having the protocol, in order to see the process in more details.
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