Korea Institute of Science and Technology (KIST), Brain Science Institute, Center for neuro-medicine, Seoul, South Korea ; School of Life Sciences and Biotechnology, Korea University, Seoul, South Korea.
PLoS One. 2013 Dec 2;8(12):e81682. doi: 10.1371/journal.pone.0081682. eCollection 2013.
Abnormal tau aggregation is a pathological hallmark of many neurodegenerative disorders and it is becoming apparent that soluble tau aggregates play a key role in neurodegeneration and memory impairment. Despite this pathological importance, there is currently no single method that allows monitoring soluble tau species in living cells. In this regard, we developed a cell-based sensor that visualizes tau self-assembly. By introducing bimolecular fluorescence complementation (BiFC) technique to tau, we were able to achieve spatial and temporal resolution of tau-tau interactions in a range of states, from soluble dimers to large aggregates. Under basal conditions, tau-BiFC cells exhibited little fluorescence intensity, implying that the majority of tau molecules exist as monomers. Upon chemically induced tau hyperphosphorylation, BiFC fluorescence greatly increased, indicating an increased level of tau-tau interactions. As an indicator of tau assembly, our BiFC sensor would be a useful tool for investigating tau pathology.
异常的 tau 聚集是许多神经退行性疾病的病理标志，越来越明显的是，可溶性 tau 聚集在神经退行性变和记忆损伤中发挥关键作用。尽管具有这种病理重要性，但目前还没有一种单一的方法可以在活细胞中监测可溶性 tau 物种。在这方面，我们开发了一种基于细胞的传感器，可以可视化 tau 自组装。通过将双分子荧光互补（BiFC）技术引入 tau，我们能够实现 tau-tau 相互作用在一系列状态下的时空分辨率，从可溶性二聚体到大型聚集物。在基础条件下，tau-BiFC 细胞表现出低荧光强度，这意味着大多数 tau 分子以单体形式存在。在化学诱导的 tau 过度磷酸化后，BiFC 荧光大大增加，表明 tau-tau 相互作用水平增加。作为 tau 组装的指标，我们的 BiFC 传感器将是研究 tau 病理学的有用工具。
