α‐Synuclein fibrils explore actin‐mediated macropinocytosis for cellular entry into model neuroblastoma neurons

IF 3.6 3区生物学 Q3 CELL BIOLOGY Traffic Pub Date : 2022-05-23 DOI:10.1111/tra.12859

Pravin Hivare, Joshna Gadhavi, D. Bhatia, Sharad Gupta

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引用次数: 1

Abstract

Alpha‐synuclein (α‐Syn), an intrinsically disordered protein (IDP), is associated with neurodegenerative disorders, including Parkinson's disease (PD or other α‐synucleinopathies. Recent investigations propose the transmission of α‐Syn protein fibrils, in a prion‐like manner, by entering proximal cells to seed further fibrillization in PD. Despite the recent advances, the mechanisms by which extracellular protein aggregates internalize into the cells remain poorly understood. Using a simple cell‐based model of human neuroblastoma‐derived differentiated neurons, we present the cellular internalization of α‐Syn PFF to check cellular uptake and recycling kinetics along with the standard endocytic markers Transferrin (Tf) marking clathrin‐mediated endocytosis (CME) and Galectin3 (Gal3) marking clathrin‐independent endocytosis (CIE). Specific inhibition of endocytic pathways using chemical inhibitors reveals no significant involvement of CME, CIE and caveolae‐mediated endocytosis (CvME). A substantial reduction in cellular uptake was observed after perturbation of actin polymerization and treatment with macropinosomes inhibitor. Our results show that α‐Syn PFF mainly internalizes into the SH‐SY5Y cells and differentiated neurons via the macropinocytosis pathway. The elucidation of the molecular and cellular mechanism involved in the α‐Syn PFF internalization will help improve the understanding of α‐synucleinopathies including PD, and further design specific inhibitors for the same.

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α -突触核蛋白原纤维探索肌动蛋白介导的巨噬细胞进入模型神经母细胞瘤神经元

α -突触核蛋白(α - Syn)是一种内在失调蛋白(IDP)，与神经退行性疾病有关，包括帕金森病(PD)或其他α -突触核蛋白病。最近的研究表明，在PD中，α - Syn蛋白原纤维以类似朊病毒的方式进入近端细胞，从而进一步形成纤维。尽管最近取得了进展，但细胞外蛋白聚集到细胞内部的机制仍然知之甚少。利用人类神经母细胞瘤衍生分化神经元的简单细胞模型，我们提出了α - Syn PFF的细胞内化，以检查细胞摄取和循环动力学，以及标准的内吞标志物转铁蛋白(Tf)标记网格蛋白介导的内吞作用(CME)和半乳糖凝聚素3 (Gal3)标记网格蛋白独立内吞作用(CIE)。化学抑制剂对内吞途径的特异性抑制表明，CME、CIE和小泡介导的内吞作用(CvME)没有明显的影响。在扰乱肌动蛋白聚合和使用巨肽体抑制剂治疗后，观察到细胞摄取的实质性减少。我们的研究结果表明，α‐Syn PFF主要通过巨噬细胞作用途径内化到SH‐SY5Y细胞和分化的神经元中。阐明参与α‐Syn PFF内化的分子和细胞机制将有助于提高对包括PD在内的α‐突触核蛋白病的理解，并进一步设计针对该疾病的特异性抑制剂。

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来源期刊

Traffic 生物-细胞生物学

CiteScore

8.10

自引率

2.20%

发文量

审稿时长

2 months

期刊介绍： Traffic encourages and facilitates the publication of papers in any field relating to intracellular transport in health and disease. Traffic papers span disciplines such as developmental biology, neuroscience, innate and adaptive immunity, epithelial cell biology, intracellular pathogens and host-pathogen interactions, among others using any eukaryotic model system. Areas of particular interest include protein, nucleic acid and lipid traffic, molecular motors, intracellular pathogens, intracellular proteolysis, nuclear import and export, cytokinesis and the cell cycle, the interface between signaling and trafficking or localization, protein translocation, the cell biology of adaptive an innate immunity, organelle biogenesis, metabolism, cell polarity and organization, and organelle movement. All aspects of the structural, molecular biology, biochemistry, genetics, morphology, intracellular signaling and relationship to hereditary or infectious diseases will be covered. Manuscripts must provide a clear conceptual or mechanistic advance. The editors will reject papers that require major changes, including addition of significant experimental data or other significant revision. Traffic will consider manuscripts of any length, but encourages authors to limit their papers to 16 typeset pages or less.