Dear all,
I'm looking for a cell line which expresses NMDA receptor transiently.
I screened HEK293 and SHSY-5Y (basal and stimulated with retinoic acid) but they don't express the receptor.
It would be useful to have an NMDAR expressing cell line cause tre transfection of NR1 and NR2A in HEK293 is difficult due to the citotoxicity mediated by the calcium.
Do you have any suggestion? Thanks.
With a quick google search, it looks like PC-12 (rat adrenal cells) express endogenous NMDAR. See this paper for reference: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1158678/pdf/jphysiol00301-0101.pdf
You could keep searching for more lines if you need human cells. I'm no expert but I didn't come across any human lines at first glance.
Alternatively, you could stably express NMDAR/NR1/NR2A in your cells with viral transduction. You could even use a doxycycline-inducible plasmid (i.e. pLIX402/403 - addgene) to minimize toxic effect from transient over-expression.
PC-12 cells express only NR1 and NR2C-D subunits (http://www.sciencedirect.com/science/article/pii/S0161813X07000745) and, unfortunately,I am interested in NR1 and NR2A-B subunits.
Moreover, in some paper researchers use different cell lines to study NMDA receptor, but they often demonstrate its presence with RT-PCR.. I want to do biochemical analysis (e.g. western blot), so this is a limit for me.
Thank you very much Stacey :)
Dear Lisa,
Adding APV to the culture media before transfection protects the HEK293 cells from NMDAR-mediated excitotoxicity.
Good luck !
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