FRET-FLIM investigation of PSD95-NMDA receptor interaction in dendritic spines; control by calpain, CaMKII and Src family kinase.
FRET-FLIM investigation of PSD95-NMDA receptor interaction in dendritic spines; control by calpain, CaMKII and Src family kinase.
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DOI：
10.1371/journal.pone.0112170
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发表时间：
2014
期刊：
影响因子：
3.7
通讯作者：
De Koninck P
中科院分区：
文献类型：
--
作者：
Doré K;Labrecque S;Tardif C;De Koninck P
突触重塑过程中突触内蛋白质相互作用的变化知之甚少，因为它们在棘中的实时监测有限。我们使用FRET FLIM方法在培养的大鼠海马神经元表达荧光标记的NMDA受体（NMDAR）和PSD 95，突触可塑性的两个重要蛋白质，检查它们之间的相互作用的调节。NMDAR刺激引起的NMDAR和PSD 95之间的年轻和成熟的神经元的脊FRET的短暂减少。CaMKII和钙蛋白酶的活性在这两个发育阶段的影响是必不可少的。同时，Src家族激酶（SFK）的抑制对年轻与成熟神经元中FRET的减少有相反的影响。我们的数据表明，协调作用的CaMKII，SFK和钙蛋白酶活性调节活动依赖性分离的PSD 95从GluN 2A或GluN 2B。最后，我们发现钙蛋白酶抑制减少了由NMDAR活性引起的棘生长，支持了PSD 95-NMDAR分离与突触重塑有关的假设。
Little is known about the changes in protein interactions inside synapses during synaptic remodeling, as their live monitoring in spines has been limited. We used a FRET-FLIM approach in developing cultured rat hippocampal neurons expressing fluorescently tagged NMDA receptor (NMDAR) and PSD95, two essential proteins in synaptic plasticity, to examine the regulation of their interaction. NMDAR stimulation caused a transient decrease in FRET between the NMDAR and PSD95 in spines of young and mature neurons. The activity of both CaMKII and calpain were essential for this effect in both developmental stages. Meanwhile, inhibition of Src family kinase (SFK) had opposing impacts on this decrease in FRET in young versus mature neurons. Our data suggest concerted roles for CaMKII, SFK and calpain activity in regulating activity-dependent separation of PSD95 from GluN2A or GluN2B. Finally, we found that calpain inhibition reduced spine growth that was caused by NMDAR activity, supporting the hypothesis that PSD95-NMDAR separation is implicated in synaptic remodeling.
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影响因子：
34.7
作者：
Lisman, John;Yasuda, Ryohei;Raghavachari, Sridhar
通讯作者：
Raghavachari, Sridhar
影响因子：
56.9
作者：
LYNCH, G;BAUDRY, M
通讯作者：
BAUDRY, M
影响因子：
4.2
作者：
Jourdi, H;Yanagihara, T;Baudry, M
通讯作者：
Baudry, M
DOI：
10.2741/s38
发表时间：
2009-06-01
期刊：
Frontiers in bioscience (Scholar edition)
影响因子：
--
作者：
Doshi S;Lynch DR
通讯作者：
Lynch DR
影响因子：
5.3
作者：
Fan, Jing;Cowan, Catherine M.;Raymond, Lynn A.
通讯作者：
Raymond, Lynn A.