Hello everyone!
In your experience, changes in the expression of which genes better represent synaptic growth and activity? I look for a marker of vesicular pool (quantity of synaptic vesicles, more precisely) and a marker of synaptic activity itself - whether the synapse is actively working or silent. I'm thinking about SV2, VAMP-associated proteins and synaptotagmin.
Do you have any suggestions?
P.S. Method - real-time PCR
Synapsin, synaptotagmin, vglut, any of these would be good to answer your question. However, if you are particularly interested size and/or number vesicles in a particular vesicular or total vesicular pool, I would suggest to go for electrophysiology. RRPs could be identified with the help of high frequency short protocols and for total vesicle pool you can take advantage of drugs like Bafilomycin A1 which blocks the refilling. For normal growth phenotype you can make use of membrane marker like HRP. Good luck
A convenient method for assessing synaptic activity is to incubate the tissue with the fluorescent molecule FM1-43, which is taken up into presynaptic terminals during vesicle recycling. This is a measure for actual neurotransmitter release.
See: Fluorescent Measurement of Synaptic Activity Using FM Dyes in Dissociated Hippocampal Cultured Neurons. Lazarenko RM, DelBove CE, Zhang Q. Bio Protoc. 2018 Jan 20;8(2)
Complementing what was said before, if FM1-43 is not working in your hands you can also try a Synaptotagmin1 uptake assay.
Hi everyone,
for synaptic activity i would suggest for c-Fos or ARK. They are the best characterized IEG for monitoring synaptic activity.
Cheers
Luca
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