Is there a way to test receptor-receptor coupling in acute brain slices (mouse)? I suspect physical interactions between an ionotropic receptor and a metabotropic receptor in the postsynaptic neuron because antogaonists of either receptor blocked the receptor-mediated responses from both kinds of receptors when the presynaptic cell is optogenetically stimulated. The presynaptic cell is known to co-release both kind of neurotransmitters that act on the aforementioned receptors on the postsynaptic cell. I hope this makes sense. I do not want to do co-IP at this stage. Thanks!
Have you herad about the proximity ligation assay?? You use the primary antobodies against the receptors you suspect physically interact. And then the secondary antibodies are linked to a DNA probe. If receptors are close enough the DNA is amplified. There are different ways to detect this amplifications.....
Have a look here:
Söderberg O, Leuchowius KJ, Gullberg M, Jarvius M, Weibrecht I, Larsson LG,
Landegren U. Characterizing proteins and their interactions in cells and tissues
using the in situ proximity ligation assay. Methods. 2008 Jul;45(3):227-32. doi:
10.1016/j.ymeth.2008.06.014. Epub 2008 Jul 11. PubMed PMID: 18620061.
Which receptors do you think are interacting? Some ligands can be fluorescently labeled, but it depends on the ligand class. Peptides are the easiest, but some others are possible, too.
GABA and dopamine receptors I suspect. How would labeling the ligands (in this case GABA and DA) work to look at the receptor interactions?
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