Interdisciplinary Research Center HALOmem, Institute of Biochemistry and Biotechnology, Charles Tanford Protein Center, Martin Luther University Halle-Wittenberg, Halle, Germany.
Institute of Anatomy and Cell Biology, Faculty of Medicine, Martin Luther University Halle-Wittenberg, Halle, Germany.
Commun Biol. 2022 Jun 7;5(1):551. doi: 10.1038/s42003-022-03478-7.
The neuroblastoma cell line SH-SY5Y is commonly employed to study neuronal function and disease. This includes cells grown under standard conditions or differentiated to neuron-like cells by administration of chemical reagents such as retinoic acid (RA) or phorbol-12-myristate-13-acetate (PMA). Even though SH-SY5Y cells are widely explored, a complete description of the resulting proteomes and cellular reorganisation during differentiation is still missing. Here, we relatively quantify the proteomes of cells grown under standard conditions and obtained from two differentiation protocols employing RA or a combination of RA and PMA. Relative quantification and KEGG pathway analysis of the proteins reveals the presence of early differentiating cells and provides a list of marker proteins for undifferentiated and differentiated cells. For characterisation of neuronal sub-types, we analyse expression of marker genes and find that RA-differentiated cells are acetylcholinergic and cholinergic, while RA/PMA-differentiated cells show high expression of acetylcholinergic and dopaminergic marker genes. In-cell cross-linking further allows capturing protein interactions in different cellular organelles. Specifically, we observe structural reorganisation upon differentiation involving regulating protein factors of the actin cytoskeleton.
神经母细胞瘤细胞系 SH-SY5Y 常用于研究神经元功能和疾病。这些细胞可以在标准条件下生长，也可以通过添加化学试剂（如视黄酸（RA）或佛波醇 12-肉豆蔻酸 13-醋酸酯（PMA））分化为类神经元细胞。尽管 SH-SY5Y 细胞已被广泛研究，但在分化过程中，对其蛋白质组和细胞重组的完整描述仍存在缺失。在这里，我们相对定量分析了在标准条件下生长的细胞和通过两种分化方案（使用 RA 或 RA 和 PMA 的组合）获得的细胞的蛋白质组。对蛋白质的相对定量和 KEGG 途径分析揭示了早期分化细胞的存在，并提供了未分化和分化细胞的标记蛋白列表。为了对神经元亚型进行特征分析，我们分析了标记基因的表达，发现 RA 分化的细胞为乙酰胆碱能和胆碱能细胞，而 RA/PMA 分化的细胞则表现出乙酰胆碱能和多巴胺能标记基因的高表达。细胞内交联进一步允许捕获不同细胞细胞器中的蛋白质相互作用。具体而言，我们观察到分化过程中的结构重组，涉及到肌动蛋白细胞骨架的调节蛋白因子。
