I have seen papers that use neuronal cultures from P0 mice and see NMDA dependent currents. For eg. see this paper:

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3129396/

Perhaps it depends on where you are isolating the neurons from? (the paper above cultured neurons from P0 striatum). I'd say give it a try! You can just whole-cell patch-clamp the cultured neurons, apply glutamate and see how much of the response is blocked by AP-5. Or do immunostaining for NMDA in either P0 brain slices or neuronal cultured from P0 animals.. 

Good luck!

Hi Anusha,

Thanks a lot for the paper ans suggestions. The paper looks convincing to me. Yes, I agree with you. A guy from my department rise up this issue and I was quite shocked. I was working on DIV 14 neurons from P0 mice pups. 

Thanks a lot.

Have a nice day!

Dear Yu,

I agree with Anusha that you need to test your cultures for FUNCTIONAL receptors.  My experience is with embryonic cortical neurons (Choi lab method) and excitatory cell death, and we would start testing the cultures at 10 DIV because there would be some variability in the time of onset of the response to NMDA.  Usually by 14 DIV they were responsive to NMDA-induced cell death, so I would think that your cultures would be responsive (depending on neuronal type).  But it would be better to be sure.

Cheers!

Jill

One other thought.. Perhaps some of the confusion arises from the presence of 'silent' synapses? As I understand, new synapses contain NMDA receptors only and AMPA receptors are recruited later as the synapse matures. But because two conditions need to be met to activate NMDA receptors - glutamate binding and removal of extracellular Mg2+ block (by a slight depolarization to about -40 mV) - it may appear as though they aren't there under normal circumstances, hence they are 'silent'. But you can again easily test this functionally by using Mg2+ free aCSF or by holding the cell at ~ -40 mV when you apply glutamate..

Hi Anusha, 

A good idea. Thanks. For the functional NMDAR, let me test either patch or Ca2+ imaging with NMDA application. If there is no response, maybe I can add glutamate or use Mg2+ free ACSF. 

Thanks for that idea.@Anusha

Yu

Hi Jill,

Thanks for the experience. Yeah, I should have a testing whether they are functional or not. I am using P0 cortical neurons to measure Ca2+ influx through NMDAR. Hope NMDA will work. 

Thanks a lot.@Jill Miotke

Yu