Effects of CaMKII inhibitor tatCN21 on activity-dependent redistribution of CaMKII in hippocampal neurons.

Effects of CaMKII inhibitor tatCN21 on activity-dependent redistribution of CaMKII in hippocampal neurons.
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DOI:
10.1016/j.neuroscience.2013.03.063
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发表时间:
2013-08-06
期刊:
影响因子:
3.3
通讯作者:
Dosemeci A
Dosemeci A
中科院分区:
医学3区
文献类型:
--
作者:
Tao-Cheng JH;Yang Y;Bayer KU;Reese TS;Dosemeci A

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TatCN21 是一种膜渗透性 CaMKII 抑制剂,源自抑制剂蛋白 CaMKIIN。 TatCN21已被用来证明CaMKII参与多种生理和病理现象,并且它还限制神经元的兴奋性毒性损伤。在这里,我们使用预嵌入免疫金电子显微镜检查 tatCN21 对培养的海马神经元中 CaMKII 重新分布的影响。在暴露于 NMDA(50 μM,2 分钟)之前,用 tatCN21(20 μM,20 分钟)孵育培养物,抑制了突触后密度(PSD)中 CaMKII 的积累和树突中 CaMKII 的聚集。在这些条件下,CaMKII 还与 PSD 附近和树突中的多核糖体形成形态不同的聚集体。这些 CaMKII-多核糖体聚集体的形成需要 tatCN21 和钙的存在,并且在暴露于高 K+ 或 NMDA 时会增强。 CaMKII-多核糖体聚集体与 20 μM tatCN21 一致形成,但与 5 μM 形成最少或根本不形成。然而,这些聚集体不是由另一种 CaMKII 抑制剂 KN93 诱导的。在洗掉 tatCN21 后 1 小时内,CaMKII-多核糖体聚集体的形成是完全可逆的。 tatCN21 的影响主要限于树突,对体细胞的影响最小。 tatCN21 对 CaMKII 分布的影响可用于剖析 CaMKII 参与细胞事件的机制。
TatCN21 is a membrane permeable CaMKII inhibitor derived from the inhibitor protein CaMKIIN. TatCN21 has been used to demonstrate the involvement of CaMKII in a variety of physiological and pathological phenomena, and it also limits excitotoxic damage in neurons. Here we use preembedding immunogold electron microscopy to examine the effect of tatCN21 on the redistribution of CaMKII in cultured hippocampal neurons. Incubation of cultures with tatCN21 (20 μM for 20 min) prior to exposure to NMDA (50 μM for 2 min) inhibited both the accumulation of CaMKII at postsynaptic densities (PSDs) and CaMKII clustering in the dendrites. Under these conditions, CaMKII also formed morphologically distinct aggregates with polyribosomes near the PSD and in dendrites. Formation of these CaMKII-polyribosome aggregates requires the presence of both tatCN21 and calcium, and was augmented upon exposure to high K+ or NMDA. CaMKII-polyribosome aggregates formed consistently with 20 μM tatCN21, but minimally or not at all with 5 μM. However, these aggregates are not induced by another CaMKII inhibitor, KN93. Formation of CaMKII-polyribosome aggregates was completely reversible within 1 h after washout of tatCN21. Effects of tatCN21 were largely restricted to dendrites, with minimal effect in the soma. The effects of tatCN21 on CaMKII distribution can be used to dissect the mechanism of CaMKII involvement in cellular events.
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