Membrane association and release of wild-type and pathological tau from organotypic brain slice cultures.
Membrane association and release of wild-type and pathological tau from organotypic brain slice cultures.
复制标题
DOI：
10.1038/cddis.2017.97
复制
发表时间：
2017-03-16
影响因子：
9
通讯作者：
Noble W
中科院分区：
文献类型：
--
作者：
Croft CL;Wade MA;Kurbatskaya K;Mastrandreas P;Hughes MM;Phillips EC;Pooler AM;Perkinton MS;Hanger DP;Noble W
病理性tau蛋白在脑中的时空传递是阿尔茨海默病的特征。可溶性和异常tau种类从健康神经元的释放在刺激神经元活性时增加。目前还不清楚控制健康神经元释放可溶性tau蛋白的机制是否与病理性tau蛋白种类扩散中所涉及的机制相同。为了开始理解这些事件，我们使用器官型脑切片培养物研究了tau蛋白的分布和释放。将来自出生后野生型和3xTg-AD小鼠的切片培养长达1个月。通过蛋白质印迹法检查亚细胞区室中的Tau分布，并使用灵敏的夹心ELISA测定释放到培养基中的Tau。我们在这里表明，3xTg-AD文化有一个加速发展的病理性tau异常，包括重新分配的tau突触和膜室。当与野生型培养物相比时，3xTg-AD切片培养物显示相对于总tau升高的基础tau释放。然而，当用氯化钾增加神经元活性时，不能进一步刺激来自3xTg-AD切片的tau释放。此外，我们报告说，在tau释放增加的条件下，脱磷酸化的膜相关tau的池增加。这些数据表明，当使用野生型和转基因小鼠脑的整合切片培养模型时，可能存在tau释放的差异模式，尽管确定tau过表达对这些发现的影响将是重要的。这些结果进一步增加了我们对神经退行性tau蛋白病中tau蛋白释放和传播的分子机制的了解。
The spatiotemporal transmission of pathological tau in the brain is characteristic of Alzheimer's disease. Release of both soluble and abnormal tau species from healthy neurons is increased upon stimulation of neuronal activity. It is not yet understood whether the mechanisms controlling soluble tau release from healthy neurons is the same as those involved in the spread of pathological tau species. To begin to understand these events, we have studied tau distribution and release using organotypic brain slice cultures. The slices were cultured from postnatal wild-type and 3xTg-AD mice for up to 1 month. Tau distribution in subcellular compartments was examined by western blotting, and tau release into culture medium was determined using a sensitive sandwich ELISA. We show here that 3xTg-AD cultures have an accelerated development of pathological tau abnormalities including the redistribution of tau to synaptic and membrane compartments. The 3xTg-AD slice cultures show elevated basal tau release relative to total tau when compared with wild-type cultures. However, tau release from 3xTg-AD slices cannot be further stimulated when neuronal activity is increased with potassium chloride. Moreover, we report that there is an increased pool of dephosphorylated membrane-associated tau in conditions where tau release is increased. These data suggest that there may be differential patterns of tau release when using integrated slice culture models of wild-type and transgenic mouse brain, although it will be important to determine the effect of tau overexpression for these findings. These results further increase our knowledge of the molecular mechanisms underlying tau release and propagation in neurodegenerative tauopathies.