N-甲基-D-天冬氨酸受体(NMDAR)是生理和病理突触可塑性的关键参与者,因为它们参与神经元传递以及学习和记忆的许多方面。不同类型的GluN2A相互作用蛋白在这些事件中的作用尚不清楚。p140Cap支架蛋白充当与精神和神经系统疾病相关的突触后复合物的枢纽,并调节突触功能,如成熟树突棘的稳定,记忆巩固,长期增强,和抑郁症。在这里,我们证明p140Cap直接结合NMDAR的GluN2A亚基并调节GluN2A相关的分子网络。的确,在p140Cap敲除雄性小鼠中,GluN2A在离体突触体和培养的海马神经元中与PSD95的相关性较小,敲除神经元中的p140Cap表达可以挽救GluN2A和PSD95的共定位。p140Cap对于招募含GluN2A的NMDAR至关重要,因此,在调节NMDAR的内在属性方面。p140Cap与突触脂质移植物(LR)和可溶性突触后膜相关,GluN2A和PSD95较少募集到p140Cap敲除雄性小鼠的突触LR中。海马神经元的g-STED显微镜检查证实,p140Cap是以活动依赖性方式将GluN2A簇嵌入LR所必需的。在突触区室中,p140Cap影响GluN2A和PSD95之间的关联,并调节GluN2A富集到LR中。总的来说,这些富含信号分子的膜结构域的这种增加导致提高的信号转导效率。重要状态我们最初显示衔接蛋白p140Cap直接结合NMDAR的GluN2A亚基并调节GluN2A相关分子网络。此外,我们首次显示p140Cap还与突触脂质移植物相关,并控制GluN2A和PSD95选择性募集到该特定区室。最后,g-STED对海马神经元的显微镜检查证实,p140Cap是以活性依赖性方式将GluN2A簇嵌入脂筏中所必需的。总的来说,我们的发现提供了p140Cap的分子和功能解剖,作为参与记忆巩固的高度动态突触网络的新活性成员,LTP和LTD已知在神经和精神疾病中发生改变。
The NMDARs are key players in both physiological and pathologic synaptic plasticity because of their involvement in many aspects of neuronal transmission as well as learning and memory. The contribution in these events of different types of GluN2A-interacting proteins is still unclear. The
p140Cap scaffold protein acts as a hub for postsynaptic complexes relevant to psychiatric and neurologic disorders and regulates synaptic functions, such as the stabilization of mature dendritic spine, memory consolidation, LTP, and LTD. Here we demonstrate that p140Cap directly binds the GluN2A subunit of NMDAR and modulates GluN2A-associated molecular network. Indeed, in
p140Cap KO male mice, GluN2A is less associated with PSD95 both in ex vivo synaptosomes and in cultured hippocampal neurons, and p140Cap expression in KO neurons can rescue GluN2A and PSD95 colocalization.
p140Cap is crucial in the recruitment of GluN2A-containing NMDARs and, consequently, in regulating NMDARs\' intrinsic properties. p140Cap is associated to synaptic lipid-raft (LR) and to soluble postsynaptic membranes, and GluN2A and PSD95 are less recruited into synaptic LR of
p140Cap KO male mice. Gated-stimulated emission depletion microscopy on hippocampal neurons confirmed that p140Cap is required for embedding GluN2A clusters in LR in an activity-dependent fashion. In the synaptic compartment,
p140Cap influences the association between GluN2A and PSD95 and modulates GluN2A enrichment into LR. Overall, such increase in these membrane domains rich in signaling molecules results in improved signal transduction efficiency.SIGNIFICANCE STATEMENT Here we originally show that the adaptor protein p140Cap directly binds the GluN2A subunit of NMDAR and modulates the GluN2A-associated molecular network. Moreover, we show, for the first time, that p140Cap also associates to synaptic lipid rafts and controls the selective recruitment of GluN2A and PSD95 to this specific compartment. Finally, gated-stimulated emission depletion microscopy on hippocampal neurons confirmed that p140Cap is required for embedding GluN2A clusters in lipid rafts in an activity-dependent fashion. Overall, our findings provide the molecular and functional dissection of p140Cap as a new active member of a highly dynamic synaptic network involved in memory consolidation, LTP, and LTD, which are known to be altered in neurologic and psychiatric disorders.