Hello! I am looking for good antibodies that help in clearly distinguish between D1- and D2-type medium spiny neurons (MSNs) in the striatum, by immunohistochemistry and immunofluorescent stainings. Which markers (also in combination) are the best to discriminate between these two MSN populations? Does anybody have any recommendations? Thank you!
back i my postdoctoral ears i used DARPP-32 antibody to stain for medium spiny neurons (see list of antibodies in this paper (PMID: 19609935), also parvalbumine+, calretinin+ and ChAT +neurons (3 types of interneurons), which are the additional of neurons in the striatum. Consider using neurotransmitter antibody as a 2nd marker. Both types D1 & D2 should be positive for GABA, and you should also be able to stain for dopamine. The 2 types clearly should have 2 different receptors D1 and D2 (not sure if you can find the antibodies differentiating between the 2) and adenylyl cyclase pathway should be active in one (D1) but not the other (D2), this may be an additioal way to differentiate
In addition to what Igor Nasonkin said; TO-PRO-3 staining could be used to identify Medium spiny neurons in striatum by their typical nuclear morphology
Thank you for all your suggestions. I used to use CTIP2 and DARPP32 marker co-localization to identify MSNs. Now I am trying to use two antibodies against D1 and D2 receptors but the staining is not the best. I am looking for good Abs against D1 and D2.
Mujittapha U. Sirajo, I didn't know about TO-PRO-3 staining. How do you discriminate MSN nuclei compared to all the other nuclei? Thank you.
- Badges
- Science method