Journal: Cell reports
Article Title: LTP expression mediated by autonomous activity of GluN2B-bound CaMKII
doi: 10.1016/j.celrep.2024.114866
Figure Lengend Snippet: LTP was induced by 2× HFS and measured by the fEPSP slopes at the CA3 to CA synapses in hippocampal slices from wild-type (WT) mice. Drug (10 μM unless noted otherwise) was added as indicated. Data show mean ± SEM. (A) When added 1 min after LTP induction, the ATP-competitive inhibitors AS397 and ruxolitinib (Ruxo) both reduced fEPSP slopes back to baseline within 60 min ( n = 7, 6, and 6 slices for control without drug, Ruxo, and AS397, respectively). (B) Significant reduction of LTP during the last 5 min of recording was observed for both AS397 and Ruxo (** p < 0.005 and *** p < 0.001 by Bonferroni multiple comparison test following one-way ANOVA). (C) When added 1 min after LTP induction, the peptide inhibitor tatCN21 (5 μM), but not the CaM-competitive inhibitor KN93, reduced fEPSP slopes back to baseline within 60 min ( n = 6 for KN93 and tatCN21). (D) During the last 5 min of recordings, only tatCN21, but not KN93, caused a significant reduction of fEPSP slope compared to the no-drug control, as also shown in the other images, and tatCN21 also caused significant reduction compared to KN93 (*** p < 0.001 and * p < 0.05 by one-way ANOVA followed by Bonferroni multiple comparison test). (E) The effect of different inhibitors (10 μM) on the autonomous activity of GluN2B-bound CaMKII was tested in vitro as illustrated in the schematic on the right. CaMKII binding was induced by Ca 2+ /CaM, which was then washed out before measuring the Ca 2+ /CaM-independent autonomous activity of the bound kinase. Negative control (−) contained no kinase and positive control (+) no inhibitor. All ATP-competitive inhibitors (AS105, AS283, and AS397) and tatCN21 completely blocked activity. KN93 had no effect on activity, and AIP reduced activity but did not block it completely. (F) When added 15 min after LTP induction, the ATP-competitive inhibitor AS397 had no apparent effect on LTP maintenance ( n = 8 slices). (G) During the last 5 min of recording, addition of ATP at 15 min after LTP induction had no significant effect on fEPSP slope compared to the control without the drug, which is also shown in the other images.
Article Snippet: KN93 , Tocris , Catalog #: 5215.
Techniques: Control, Comparison, Activity Assay, In Vitro, Binding Assay, Negative Control, Positive Control, Blocking Assay