α-Synuclein (αSyn) Preformed Fibrils Induce Endogenous αSyn Aggregation, Compromise Synaptic Activity and Enhance Synapse Loss in Cultured Excitatory Hippocampal Neurons
α-Synuclein (αSyn) Preformed Fibrils Induce Endogenous αSyn Aggregation, Compromise Synaptic Activity and Enhance Synapse Loss in Cultured Excitatory Hippocampal Neurons
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DOI：
10.1523/jneurosci.0060-19.2019
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发表时间：
2019-06-26
影响因子：
5.3
通讯作者：
Lee, Virginia M-Y
中科院分区：
文献类型：
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作者：
Wu, Qihui;Takano, WHajime;Lee, Virginia M-Y
突触核病的特征是不溶的α-突触核蛋白(α-突触核蛋白)积聚。为了测试α-Syn聚集体是否调节突触活动，我们使用了最近在初级神经元中开发的一个模型来诱导α-Syn病理。我们证实，重组人α-Syn产生的预形成纤维(PFF)以时间和剂量依赖的方式损害突触活性，这些缺陷的大小与培养的两性小鼠兴奋性海马神经元α-Syn病理的形成有关。显著的是，从全细胞膜片钳吸管中急性被动输注α-Syn-pff可在10min内降低mEPSC频率，并在1d内诱发α-Syn病变。此外，通过直接向培养液中加入错误折叠的α-Syn包涵体，突触标志物的共定位和树突棘的动态变化显著地受到影响，但处理后7d，神经元的存活率不受影响。我们的数据表明，神经元内的α-突触纤维损害了突触发生的启动及其生理功能，从而提示靶向突触功能障碍在突触核病中可能提供一个有前途的治疗方向。
Synucleinopathies are characterized by the accumulation of insoluble alpha-synuclein (alpha Syn). To test whether alpha Syn aggregates modulate synaptic activity, we used a recently developed model in primary neurons for inducing alpha Syn pathology. Wedemonstrated that preformed fibrils (PFFs) generated with recombinant human alpha Syn compromised synaptic activity in a time-and dose-dependent manner and that the magnitude of these deficits correlated with the formation of alpha Syn pathology in cultured excitatory hippocampal neurons from both sexes of mice. Remarkably, acute passive infusion of alpha Syn PFFs from whole-cell patch-clamp pipette decreased mEPSC frequency within 10 min followed by induction of alpha Syn pathology within 1 d. Moreover, by direct addition of alpha Syn PFFs into culture medium, the formation of misfolded alpha Syn inclusions dramatically compromised the colocalization of synaptic markers and altered dynamic changes of dendritic spines, but the viability of neurons was not affected up to 7 d post-treatment with alpha Syn PFFs. Our data indicate that intraneuronal alpha Syn fibrils impaired the initiation of synaptogenesis and their physiological functions, thereby suggesting that targeting synaptic dysfunction in synucleinopathies may provide a promising therapeutic direction.