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Alpha-synuclein promotes early neurite outgrowth in cultured primary neurons

  • Neurology and Preclinical Neurological Studies - Original Article
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Abstract

We previously showed that alpha-synuclein (α-Syn), a protein implicated in the pathogenesis of several neurodegenerative diseases, is a microtubule-associated protein (MAP), facilitating the polymerization of tubulin into microtubules. Therefore, we hypothesized that α-Syn might promote neurite outgrowth, a process that requires microtubule assembly. To test this hypothesis, recombinant human wild type (WT) and mutant (A30P and A53T) α-Syn proteins were added to cultured primary rat cortical neurons, and their effects on early neurite outgrowth were observed. The WT and mutant α-Syn proteins entered the neurons after 1–4 h of incubation. However, a significant increase in neurite outgrowth was observed only in neurons treated with WT α-Syn. MES23.5 dopaminergic neuronal cells overexpressing WT α-Syn also exhibited enhanced neurite outgrowth, indicating that the ability of α-Syn to promote neurite outgrowth was not due to a direct action on the cell membrane or by the membrane translocation process. Co-immunoprecipitation demonstrated that the recombinant human α-Syn was bound to tubulin. In addition, the α-Syn-treated neurons displayed increased levels of polymerized tubulin. Because α-Syn’s MAP functionality is mediated by specific domains, we generated N-terminal (a.a. 1–65), non-amyloid-β (non-Aβ) component (NAC) (a.a. 61–95) and C-terminal (a.a. 96–140) fragments and added them to the primary neurons. After 1–4 h of incubation, the various α-Syn fragments had entered the neurons. However, only the NAC and C-terminal fragments, which have been previously shown to mediate MAP functionality, promoted neurite outgrowth. These results suggest that α-Syn promotes neurite outgrowth by facilitating the polymerization of tubulin into microtubules.

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Acknowledgments

This work was supported by grants from the National Basic Research Program (‘‘973” Program) of China (2011CB504101), National High Technology Research and Development Program (‘‘863” Program) of China (2006AA02A408), National Natural Science Foundation of China (30270482, 30271437, 30430280 and 81071014), Natural Science Foundation of Beijing (303-01-003-0039, 7022011, 7122035, and 7102076), Funding Project for Academic Human Resources Development in the Institutions of Higher Learning under the Jurisdiction of Beijing Municipality (PHR200907113), Japan Society for the Promotion of Science (C23500408), and the “Twelfth Five-Year Plan” Science and Technology Research Projects of Jilin Province Education Department [2012]-137.

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We declare that there are no conflicts of interest.

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Authors and Affiliations

  1. Department of Neurobiology, Beijing Institute of Geriatrics, Xuanwu Hospital of China Capital Medical University, No. 45 Changchun Street, Beijing, 100053, China

    Guangwei Liu, Peng Wang, Xin Li, Yaohua Li, Shengli Xu, Kenji Uéda, Piu Chan & Shun Yu

  2. Key Laboratory of Neurodegenerative Diseases (Capital Medical University), Ministry of Education, Beijing, 100053, China

    Guangwei Liu, Peng Wang, Xin Li, Yaohua Li, Shengli Xu, Piu Chan & Shun Yu

  3. Department of Dementia and Higher Brain Function, Tokyo Metropolitan Institute of Medical Science, Tokyo, 156-8506, Japan

    Kenji Uéda

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  1. Guangwei Liu
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Correspondence to Shun Yu.

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G. Liu and P. Wang are co-first authors. K. Uéda is co-principal investigator.

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Liu, G., Wang, P., Li, X. et al. Alpha-synuclein promotes early neurite outgrowth in cultured primary neurons. J Neural Transm 120, 1331–1343 (2013). https://doi.org/10.1007/s00702-013-0999-8

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