Activity-dependent release of phosphorylated human tau from Drosophila neurons in primary culture.
Activity-dependent release of phosphorylated human tau from Drosophila neurons in primary culture.
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DOI：
10.1016/j.jbc.2021.101108
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发表时间：
2021-10
期刊：
影响因子：
--
通讯作者：
Lee D
中科院分区：
文献类型：
--
作者：
Ismael S;Sindi G;Colvin RA;Lee D
神经元活动可以增强tau蛋白的释放，从而加速tau蛋白病。这种活性依赖性tau释放可用于研究阿尔茨海默病（AD）中tau病理学的进展，因为过度磷酸化的tau涉及AD发病机制和相关的tau病变。然而，我们对调节活性依赖性tau蛋白从神经元释放的机制以及tau蛋白磷酸化在调节活性依赖性tau蛋白释放中所起的作用的理解仍然是初步的。在这项研究中，原代培养的表达人tau蛋白（hTau）的果蝇神经元被用来研究活性依赖性tau蛋白的释放。我们发现50 mM KCl处理1 h显著增加了hTau的释放。使用光遗传学技术观察到类似水平的释放，其中使用蓝光（470 nm）刺激遗传靶向的神经元30分钟。我们的结果表明，与野生型hTau相比，磷酸抗性hTauS 11 A的活性依赖性释放减少。相反，磷酸化模拟hTauE 14的释放在活化后增加。我们发现释放的hTau在其富含脯氨酸的和C-末端结构域中被磷酸化，使用磷酸化位点特异性tau抗体（例如，AT8）。细胞裂解物中或条件培养基免疫纯化后总hTau或磷酸化hTau可检测水平的倍数变化与光刺激后磷酸化hTau的优先释放一致。这项研究建立了一个很好的模型来研究活性依赖性hTau释放的机制，并更好地理解磷酸化tau释放在AD发病机制中的作用，因为它涉及与神经元活性增加相关的神经变性早期阶段的改变。
Neuronal activity can enhance tau release and thus accelerate tauopathies. This activity-dependent tau release can be used to study the progression of tau pathology in Alzheimer's disease (AD), as hyperphosphorylated tau is implicated in AD pathogenesis and related tauopathies. However, our understanding of the mechanisms that regulate activity-dependent tau release from neurons and the role that tau phosphorylation plays in modulating activity-dependent tau release is still rudimentary. In this study, Drosophila neurons in primary culture expressing human tau (hTau) were used to study activity-dependent tau release. We found that hTau release was markedly increased by 50 mM KCl treatment for 1 h. A similar level of release was observed using optogenetic techniques, where genetically targeted neurons were stimulated for 30 min using blue light (470 nm). Our results showed that activity-dependent release of phosphoresistant hTauS11A was reduced when compared with wildtype hTau. In contrast, release of phosphomimetic hTauE14 was increased upon activation. We found that released hTau was phosphorylated in its proline-rich and C-terminal domains using phosphorylation site-specific tau antibodies (e.g., AT8). Fold changes in detectable levels of total or phosphorylated hTau in cell lysates or following immunopurification from conditioned media were consistent with preferential release of phosphorylated hTau after light stimulation. This study establishes an excellent model to investigate the mechanism of activity-dependent hTau release and to better understand the role of phosphorylated tau release in the pathogenesis of AD since it relates to alterations in the early stage of neurodegeneration associated with increased neuronal activity.
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影响因子：
4.8
作者：
Merezhko M;Uronen RL;Huttunen HJ
通讯作者：
Huttunen HJ
影响因子：
4.3
作者：
Kimura T;Sharma G;Ishiguro K;Hisanaga SI
通讯作者：
Hisanaga SI
影响因子：
4.6
作者：
Mohamed NV;Plouffe V;Rémillard-Labrosse G;Planel E;Leclerc N
通讯作者：
Leclerc N
影响因子：
5.3
作者：
Kosmidis, Stylianos;Grammenoudi, Sofia;Skoulakis, Efthimios M. C.
通讯作者：
Skoulakis, Efthimios M. C.
磷酸钙钙调神经蛋白会影响tau代谢。
影响因子：
4.2
作者：
Karch CM;Jeng AT;Goate AM
通讯作者：
Goate AM