Effects of BTS (N-benzyl-p-toluene sulphonamide), an inhibitor for myosin-actin interaction, on myofibrillogenesis in skeletal muscle cells in culture
Effects of BTS (N-benzyl-p-toluene sulphonamide), an inhibitor for myosin-actin interaction, on myofibrillogenesis in skeletal muscle cells in culture
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DOI：
10.2108/zsj.23.969
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发表时间：
2006-11-01
影响因子：
0.9
通讯作者：
Obinata, Takashi
中科院分区：
文献类型：
--
作者：
Kagawa, Maiko;Sato, Naruki;Obinata, Takashi
肌动蛋白丝以正确的极性和六边形排列围绕肌球蛋白丝排列，形成交叉条纹结构。据推测，这种肌球蛋白-肌动蛋白相互作用在肌原纤维形成的初始阶段很重要。先前已证明，肌动蛋白-肌球蛋白相互作用的抑制剂 BDM（2,3-丁二酮单肟）可抑制培养物中肌肉细胞中肌原纤维的形成。然而，进一步的研究表明，BDM 还对活细胞产生一些额外的影响。在本研究中，我们通过应用肌球蛋白 ATP 酶和肌动蛋白-肌球蛋白相互作用的更特异性抑制剂 BTS（N-苄基-对甲苯磺酰胺），进一步研究了鸡胚骨骼肌原代培养物中肌动蛋白-肌球蛋白相互作用在肌原纤维组装中的作用。通过免疫细胞化学方法对融合后的肌管应用 BTS 来检查肌原纤维蛋白肌节结构的组装。添加 BTS (1050 μM) 显着抑制肌动蛋白和肌球蛋白组织成横纹结构。 BTS 还会干扰 α-肌动蛋白、C 蛋白（或 MyBP-C）和连接蛋白（或肌联蛋白）组织成有序的条纹结构，但敏感性较低。此外，当将在BTS存在下培养的肌管转移到对照培养基中时，在2-3天内形成肌节结构，表明BTS对肌管的抑制作用是可逆的。这些结果表明肌动蛋白-肌球蛋白相互作用在肌原纤维形成过程中起着至关重要的作用。
Actin filaments align around myosin filaments in the correct polarity and in a hexagonal arrangement to form cross-striated structures. It has been postulated that this myosin-actin interaction is important in the initial phase of myofibrillogenesis. It was previously demonstrated that an inhibitor of actin-myosin interaction, BDM (2,3-butanedione monoxime), suppresses myofibril formation in muscle cells in culture. However, further study showed that BDM also exerts several additional effects on living cells. In this study, we further examined the role of actin-myosin interaction in myofibril assembly in primary cultures of chick embryonic skeletal muscle by applying a more specific inhibitor, BTS (N-benzyl-p-toluene sulphonamide), of myosin ATPase and actin-myosin interaction. The assembly of sarcomeric structures from myofibrillar proteins was examined by immunocytochemical methods with the application of BTS to myotubes just after fusion. Addition of BTS (1050 mu M) significantly suppressed the organization of actin and myosin into cross-striated structures. BTS also interfered in the organization of alpha-actinin, C-protein (or MyBP-C), and connectin (or titin) into ordered striated structures, though the sensitivity was less. Moreover, when myotubes cultured in the presence of BTS were transferred to a control medium, sarcomeric structures were formed in 2-3 days, indicating that the inhibitory effect of BTS on myotubes is reversible. These results show that actin-myosin interaction plays a critical role in the process of myofibrillogenesis.