In vivo aggregation of presynaptic alpha-synuclein is not influenced by its phosphorylation at serine-129.
In vivo aggregation of presynaptic alpha-synuclein is not influenced by its phosphorylation at serine-129.
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DOI：
10.1016/j.nbd.2021.105291
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发表时间：
2021-05
影响因子：
6.1
通讯作者：
Unni, Vivek K.
中科院分区：
文献类型：
--
作者：
Weston, Leah J.;Cook, Zoe T.;Stackhouse, Teresa L.;Sal, Mehtab K.;Schultz, Baergen I.;Tobias, Zachary J. C.;Osterberg, Valerie R.;Brockway, Nicole L.;Pizano, Saheli;Glover, Greta;Weissman, Tamily A.;Unni, Vivek K.
关键词：
α-突触核蛋白的异常聚集是帕金森病和其他神经退行性疾病的关键分子特征。触发α-突触核蛋白聚集的确切机制尚不清楚，也不知道聚集在疾病发病机制中所起的作用。在这里，我们使用活体斑马鱼模型来表达几种不同形式的人α-突触核蛋白，并测量其在突触前终末的聚集。我们发现，绿色荧光蛋白标记的人α突触核蛋白可以在斑马鱼神经元中表达，正常定位于突触前终末，并在丝氨酸-129处发生磷酸化，就像在哺乳动物神经元中一样。斑马鱼系统的视觉优势使动态体内成像能够研究α-突触核蛋白，包括使用光漂白后荧光恢复技术来探测蛋白质的迁移率。这些实验揭示了三个不同的α-突触核蛋白末端池，它们的迁移率不同，可能代表了聚集和非聚集蛋白质的不同亚群。人的α-突触核蛋白被内源性的斑马鱼Polo样激酶活性所磷酸化，在整个轴突中有一个异质性的神经元群体，其磷酸化程度很低或很广泛。对丝氨酸-129的药理和遗传操作都表明，该位点的α-突触核蛋白的磷酸化不会显著影响其流动性。这表明丝氨酸-129单独的磷酸化并不促进α-突触核蛋白的聚集。总之，我们的结果表明，人类α-突触核蛋白可以在斑马鱼中表达和定量测量，并且与疾病相关的翻译后修饰发生在神经元中。斑马鱼模型为测量和操纵α-突触核蛋白的功能和聚集，以及为神经退行性疾病开发新的治疗方法提供了一个强大的体内系统。
Abnormal aggregation of the α-synuclein protein is a key molecular feature of Parkinson’s disease and other neurodegenerative diseases. The precise mechanisms that trigger α-synuclein aggregation are unclear, and it is not known what role aggregation plays in disease pathogenesis. Here we use an in vivo zebrafish model to express several different forms of human α-synuclein and measure its aggregation in presynaptic terminals. We show that human α-synuclein tagged with GFP can be expressed in zebrafish neurons, localizing normally to presynaptic terminals and undergoing phosphorylation at serine-129, as in mammalian neurons. The visual advantages of the zebrafish system allow for dynamic in vivo imaging to study α-synuclein, including the use of fluorescence recovery after photobleaching (FRAP) techniques to probe protein mobility. These experiments reveal three distinct terminal pools of α-synuclein with varying mobility, likely representing different subpopulations of aggregated and non-aggregated protein. Human α-synuclein is phosphorylated by an endogenous zebrafish Polo-like kinase activity, and there is a heterogeneous population of neurons containing either very little or extensive phosphorylation throughout the axonal arbor. Both pharmacological and genetic manipulations of serine-129 show that phosphorylation of α-synuclein at this site does not significantly affect its mobility. This suggests that serine-129 phosphorylation alone does not promote α-synuclein aggregation. Together our results show that human α-synuclein can be expressed and measured quantitatively in zebrafish, and that disease-relevant post-translational modifications occur within neurons. The zebrafish model provides a powerful in vivo system for measuring and manipulating α-synuclein function and aggregation, and for developing new treatments for neurodegenerative disease.
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影响因子：
168.9
作者：
Chartier-Harlin, MC;Kachergus, J;Destée, A
通讯作者：
Destée, A
DOI：
10.1073/pnas.0711053105
发表时间：
2008-01-15
影响因子：
11.1
作者：
Gorbatyuk, Oleg S.;Li, Shoudong;Muzyczka, Nicholas
通讯作者：
Muzyczka, Nicholas
影响因子：
168.9
作者：
Ibáñez, P;Bonnet, AM;Brice, A
通讯作者：
Brice, A
影响因子：
6.1
作者：
Febbraro, Fabia;Sahin, Gurdal;Romero-Ramos, Marina
通讯作者：
Romero-Ramos, Marina
DOI：
10.1107/s1744309108019623
发表时间：
2008-08-01
影响因子：
0.9
作者：
Elling, Robert A.;Fucini, Raymond V.;Romanowski, Michael J.
通讯作者：
Romanowski, Michael J.