Hello,
I've been trying to measure NMDA currents in the BLA. I've been using magnesium free acsf (122 NaCl, 2 KCl, 1.25 KH2PO4, 26 NaHCO3, 2.5 CaCl2, 10 D-glucose with nbqx (5uM) and bicuculine (5uM) and stimulating external capsule while principal cell was held at -70 mV, but I can't get any baseline and they are very unstable. I was wondering about changing it to normal acsf with 1.3 MgSO4 and measure it at +40 mV or maybe try to stimulate locally or in central amygdala? Do you have any good practice to measure NMDA currents?
I would be very grateful for answers.
I would test to make sure you're getting stable EPSCs before trying to measure the NMDA component of the EPSC. If you're not reliably evoking stable EPSCs, playing with the conditions to isolate the NMDA currents will just be frustrating. Using a Cs based internal solution with normal aCSF and measuring at +40 mV would be one way, and would probably be the easiest. That way, you can ensure you have a stable EPSC at -60mV and then block AMPA-R's with NBQX and shift to +40mV to measure the NMDA currents. Alternatively, reducing the Mg in your bath and measuring at -60 mV is another method people have used. You might also consider adding a low concentration of glycine to the bath to help facilitate the NMDA currents, even though it shouldn't be limiting.
- Badges
- Science topic
- Similar topics
- Methodology
- Research Methodology
- Research Papers