As shown Olaparib price in Figure 4A, P0 deletion of either GluN1 or both GluN2A and GluN2B results in a complete elimination of NMDAR-EPSCs in paired CA1 pyramidal neurons. Single-gene deletion of GluN2A had

no effect on NMDAR-EPSC amplitude (Figure 4B), while GluN2B deletion resulted in an approximately 40% reduction in peak EPSC amplitude (Figure 4B). Given the differences in decay kinetics between GluN2A and GluN2B diheteromeric receptors, these differences in peak amplitude would be expected to have large impacts on total charge transfer per EPSCs. Indeed, approximately 1.8-fold more charge was transferred per NMDAR-EPSC in ΔGluN2A cells than control cells (Figure 4C). Conversely, the total charge transfer per NMDAR-EPSCs from ΔGluN2B cells was only about 25% that of control cells (Figure 4C). Due to the significant differences in NMDAR-EPSCs between ΔGluN2A and ΔGluN2B cells, we examined the effects of GluN2 subunit deletion on AMPAR-EPSCs as a means of assessing synaptic strength and function. We have recently shown that late embryonic deletion of GluN1 in CA1 pyramidal neurons increases AMPAR-EPSCs and enhances the number of functional synapses (Adesnik et al., 2008) via a homeostatic-like

VX-770 chemical structure mechanism (Lu et al., 2011). Similarly, we show here that postnatal deletion of either GluN1 or simultaneous deletion of both GluN2A and GluN2B also results in a significant increase in AMPAR-EPSCs (Figure 5A). Surprisingly, deletion of either GluN2A or GluN2B individually also resulted in a similar increase in AMPAR-EPSCs (Figure 5B). As none of the genetic deletions

affected the paired-pulse ratio Ridaforolimus (Deforolimus, MK-8669) (Figure 5C), a measure of transmitter release probability, these effects are likely to be postsynaptic in origin. Furthermore, we recently demonstrated that the potentiation of AMPARs after deletion of GluN1 requires the GluA2 subunit (Lu et al., 2011). In agreement, there were no changes in AMPAR-EPSC rectification, a measure of the GluA2 content of AMPARs (Figure 5D), after deletion of GluN2A, GluN2B or both, suggesting that AMPARs trafficked to synapses contain the GluA2 subunit. Given the unexpected finding that deletion of either GluN2A or GluN2B results in the potentiation of AMPAR-EPSCs, we next asked whether these manipulations may be increasing AMPAR responses by different mechanisms. For instance, the increase in synaptic transmission could be due to enhanced synaptic strength at individual synapses or to a greater number of functional synaptic inputs. To test this, we measured AMPA receptor-mediated, action potential-independent, miniature excitatory postsynaptic currents (mEPSCs) in neighboring Cre-expressing and control cells.