Home > Internet > The Entire Study Behind AZD0156

The Entire Study Behind AZD0156

Added: (Tue Mar 13 2018)

Pressbox (Press Release) - No significant difference in the relative amount of GluK5 (or GluK2a) exported to the plasma membrane was observed when the phospho-null (GluK5AAA) or the phospho-mimetic (GluK5DDD) mutants of GluK5 were expressed with GluK2a (Figure 3C). When assembled with GluK2b, surface expression of GluK5wt and GluK5AAA was slightly lower than with GluK2a (Figure 3C). In contrast, the phospho-mimetic mutant GluK5DDD slightly enhanced surface expression of the GluK2b/GluK5 heteromer (both surface GluK2b and GluK5DDD increase) (P<0.05) (Figure 3C). These experiments are in general consistent with the electrophysiological Crenolanib analysis of recombinant GluK2/GluK5 receptors expressed in HEK 293 cells (Supplementary Figure S4). The impact of GluK5 mutations on the surface expression of myc�CGluK5wt was analysed in cultured hippocampal neurons derived from GluK2?/? x GluK5?/? mice (Figure 4A and B). Following co-transfection of myc�CGluK5 with either GluK2a or GluK2b, extracellular and total expression of GluK5 were detected CPI-0610 concentration with anti-myc antibodies (Figure 4A). Myc�CGluK5wt and myc�CGluK5AAA co-expressed with GluK2a or GluK2b were present at the plasma membrane in comparable amounts (Figure 4B) to that reported in WT hippocampal neurons (Nasu-Nishimura et al, 2006). Expression of myc�CGluK5DDD with either GluK2a or GluK2b subunits markedly increased surface expression of GluK2/GluK5 receptors (P<0.001). To confirm the role of CaMKII-dependent phosphorylation of GluK5wt, neurons from KO mice were co-transfected with GluK2a/myc�CGluK5wt or GluK2b/myc�CGluK5wt and either wt CaMKII or a truncated constitutively active CaMKII (tCaMKII). TCaMKII markedly increased surface expression of GluK5 when co-assembled with either GluK2a or ATR inhibitor GluK2b (P<0.001) (Figure 4C). As a control, expression of tCaMKII did not change the plasma membrane localization of GluK5AAA, which cannot be phosphorylated, when expressed with either GluK2a or GluK2b (Figure 4C). Interestingly, the levels reached for the amount of surface expressed GluK5wt with tCaMKII are comparable to those obtained with myc�CGluK5DDD, indicating that this phospho-mimetic mutant represents a good approximation of GluK5 subunit phosphorylated by CaMKII. The fact that CaMKII markedly promotes surface expression of GluK5-containing KARs may appear at odds with our initial observation that CaMKII is involved in the depression of synaptic KARs in a GluK5-dependent manner. We thus measured the amount of myc�CGluK5 co-localized with Homer1C�CGFP, a marker of postsynaptic densities, in dendritic spines of cultured hippocampal neurons (Figure 4D�CF). Whereas the levels of myc�CGluK5wt and myc�CGluK5AAA were comparable, the levels of phospho-mimetic myc�CGluK5DDD were markedly reduced at dendritic spines (P<0.0001) (Figure 4F).

Submitted by:
Disclaimer: Pressbox disclaims any inaccuracies in the content contained in these releases. If you would like a release removed please send an email to remove@pressbox.co.uk together with the url of the release.