Department of Basic Veterinary Science, Joint Faculty of Veterinary Medicine, Kagoshima University, Kagoshima, Japan.
Department of Pharmacology, Faculty of Veterinary Science, Bangladesh Agricultural University, Mymensingh, Bangladesh.
J Vet Med Sci. 2024 Nov 1;86(11):1136-1144. doi: 10.1292/jvms.24-0276. Epub 2024 Sep 27.
Myristoylated alanine-rich protein kinase C substrate (MARCKS) plays crucial roles in neuronal functions and differentiation. However, specific effects of the myristoylated N-terminal sequence (MANS) peptide, a widely used MARCKS modulator comprising the initial 24 amino acids of MARCKS, on neuronal cells remain unclear. Therefore, in this study, we aimed to examine the effects and action mechanisms of the MANS peptide on SH-SY5Y human neuroblastoma cells, which served as the in vitro neuronal cell models. MANS treatment of SH-SY5Y cells resulted in significant neurite outgrowth within 24 hr, which was as prominent as that induced by seven days of treatment with all-trans retinoic acid, the most common agent used to induce SH-SY5Y cell differentiation. Levels of synaptophysin, a neuronal marker protein, were significantly increased in the MANS peptide-treated cells. Additionally, increased MARCKS levels and decreased MARCKS phosphorylation were observed in MANS peptide-treated cells. Notably, neurite outgrowth induced by the MANS peptide was significantly reduced in MARCKS-knocked-down cells. Overall, these results suggest the MANS peptide as a novel agent for SH-SY5Y cell differentiation, particularly for the analysis of MARCKS functions.
肉豆蔻酰化丙氨酸丰富蛋白激酶 C 底物（MARCKS）在神经元功能和分化中发挥着至关重要的作用。然而，广泛用于 MARCKS 调节剂的起始 24 个氨基酸组成的肉豆蔻酰化 N 端序列（MANS）肽对神经元细胞的具体影响仍不清楚。因此，在这项研究中，我们旨在研究 MANS 肽对 SH-SY5Y 人神经母细胞瘤细胞的影响及其作用机制，后者作为体外神经元细胞模型。MANS 处理 SH-SY5Y 细胞在 24 小时内导致明显的神经突生长，与全反式视黄酸（最常用于诱导 SH-SY5Y 细胞分化的试剂）处理 7 天诱导的效果一样显著。MANS 肽处理的细胞中突触小泡蛋白（一种神经元标记蛋白）的水平显著增加。此外，在 MANS 肽处理的细胞中观察到 MARCKS 水平增加和 MARCKS 磷酸化减少。值得注意的是，MARCKS 敲低细胞中的神经突生长由 MANS 肽诱导的明显减少。总体而言，这些结果表明 MANS 肽是一种用于 SH-SY5Y 细胞分化的新型试剂，特别适用于 MARCKS 功能的分析。
