Research Center of the University of Montreal Hospital (CRCHUM), Montréal, Canada.
Research Center of the University of Montreal Hospital (CRCHUM), Montréal, Canada; Département de Neurosciences, Faculté de Médecine, Université de Montréal, Montréal, Canada.
J Biol Chem. 2020 Dec 18;295(51):17827-17841. doi: 10.1074/jbc.RA120.013553.
In Alzheimer's disease (AD), tau, a microtubule-associated protein (MAP), becomes hyperphosphorylated, aggregates, and accumulates in the somato-dendritic compartment of neurons. In parallel to its intracellular accumulation in AD, tau is also released in the extracellular space, as revealed by its increased presence in cerebrospinal fluid (CSF). Consistent with this, recent studies, including ours, have reported that neurons secrete tau, and several therapeutic strategies aim to prevent the intracellular tau accumulation. Previously, we reported that late endosomes were implicated in tau secretion. Here, we explore the possibility of preventing intracellular tau accumulation by increasing tau secretion. Using neuronal models, we investigated whether overexpression of the vesicle-associated membrane protein 8 (VAMP8), an R-SNARE found on late endosomes, could increase tau secretion. The overexpression of VAMP8 significantly increased tau secretion, decreasing its intracellular levels in the neuroblastoma (N2a) cell line. Increased tau secretion by VAMP8 was also observed in murine hippocampal slices. The intracellular reduction of tau by VAMP8 overexpression correlated to a decrease of acetylated tubulin induced by tau overexpression in N2a cells. VAMP8 staining was preferentially found on late endosomes in N2a cells. Using total internal reflection fluorescence (TIRF) microscopy, the fusion of VAMP8-positive vesicles with the plasma membrane was correlated to the depletion of tau in the cytoplasm. Finally, overexpression of VAMP8 reduced the intracellular accumulation of tau mutants linked to frontotemporal dementia with parkinsonism and α-synuclein by increasing their secretion. Collectively, the present data indicate that VAMP8 could be used to increase tau and α-synuclein clearance to prevent their intracellular accumulation.
在阿尔茨海默病(AD)中,微管相关蛋白(MAP)tau 过度磷酸化、聚集并在神经元的体树突区累积。与 AD 中其在细胞内的积累平行的是,tau 也在细胞外空间释放,这一点从其在脑脊液(CSF)中的含量增加中可以得到证实。与这一发现一致的是,最近的研究包括我们的研究报告称,神经元会分泌 tau,并且有几种治疗策略旨在防止细胞内 tau 积累。以前,我们报告说晚期内体参与 tau 的分泌。在这里,我们探索通过增加 tau 分泌来防止细胞内 tau 积累的可能性。使用神经元模型,我们研究了过度表达囊泡相关膜蛋白 8(VAMP8)是否可以增加 tau 分泌。VAMP8 的过度表达显著增加了 tau 的分泌,降低了神经母细胞瘤(N2a)细胞系中 tau 的细胞内水平。在小鼠海马切片中也观察到 VAMP8 增加 tau 的分泌。VAMP8 过表达导致 tau 在内质网中的减少与 tau 过表达在 N2a 细胞中诱导的乙酰化微管减少相关。VAMP8 染色在 N2a 细胞中主要存在于晚期内体上。使用全内反射荧光(TIRF)显微镜,VAMP8 阳性囊泡与质膜的融合与细胞质中 tau 的耗竭相关。最后,VAMP8 的过表达通过增加它们的分泌来减少与额颞叶痴呆伴帕金森病和α-突触核蛋白相关的 tau 突变体的细胞内积累。总之,这些数据表明 VAMP8 可用于增加 tau 和 α-突触核蛋白的清除,以防止其细胞内积累。