Myosin regulation in the migration of tumor cells and leukocytes within a three-dimensional collagen matrix

The migration of cells is a complex regulatory process which results in the generation of motor forces through the reorganization of the cytoskeleton. Here we present a comparative study of the expression and involvement of myosin in the regulation of the physiological migration of leukocytes and the pathological migration of tumor cells. We show that the involvement of myosin in the migration is distinct in these two cell types. In leukocytes, the activity of non-muscle myosin II is essential for both the spontaneous (matrix-induced) migration and the migration induced by ligands to G protein-coupled receptors, i.e. chemokines and neurotransmitters. In contrast, spontaneous tumor cell migration is largely independent of non-muscle myosin II activity, whereas the norepinephrine-induced migration is completely inhibited by either direct inhibition of non-muscle myosin II or of the kinases phosphorylating the myosin light chain, namely ROCK or the calcium/calmodulin-dependent myosin light-chain kinase.Received 31 August 2004; accepted 26 October 2004     

PH值对应用肌球蛋白 ATP酶法进行梭内肌纤维分型的影响

探讨酸性和碱性预孵育处理，对应用肌球蛋白ATP酶法进行梭内肌纤维分型的影响。采用肌球蛋白ATP酶法。PH4．3和PH4．6预孵育液孵育后，梭内肌纤维中的核袋纤维染色呈阳性，核链纤维染色相对较浅；PH9．4预孵育后，核袋与核链纤维均呈阳性或强阳性；PH10．4预孵育后，核袋1纤维呈阴性，核袋2纤维呈阳性，核链纤维呈强阳性。结论是：用mATP酶法研究梭内肌纤维的分型时，预孵育液的PH值对分型结果有显著影响，应当说明并严格把握预孵育液的PH值。     

家兔在发育过程中外侧腓肠肌内MyHCs变化与诱发电位的关系

 为研究在生后不同年龄组家兔外侧腓肠肌各亚体肌纤维MyHCs变化与诱发电位的关系,探讨生后发育期间肌纤维MyHCs组成和作用差异与表型之间的相关联系,采用电生理记录仪结合十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS PAGE）检测外侧腓肠肌各亚体。结果表明中间亚体在刺激时各年龄组的峰值电压基本相同,而外侧亚体和内侧亚体分别有些变化。同时发现成年家兔的持续时间都较幼年的长,这可能与各亚体的肌纤维型构成比例有很大关系。外侧腓肠肌各亚体的肌球蛋白重链异构体（MyHCs）电泳条带分别显示MyHCsⅡa、Ⅱd（或Ⅱx）、Ⅱb、Ⅰ共4种,对应于骨骼肌纤维表型ⅡA型（FOG型）、ⅡX（FO型）、ⅡB型（FG型）及Ⅰ型（SO型）4类。     

肌球蛋白工作循环模型

目的基于肌肉的特殊结构,给出肌球蛋白工作循环的四态模型,研究肌肉中力与位移的关系。方法利用化学动力学方法及其非平衡稳态解,从物理学的角度讨论肌肉中力与位移的关系。结果力与位移的关系与实验结果基本吻合。结论此模型可用于讨论肌肉的力与位移的关系,而且化学动力学方法适用于研究本文体系。     

Coupling mechanism of multi-force interactions in the myosin molecular motor

The dynamics of the myosin molecular motor as it binds to actin filaments during muscle contraction are still not clearly understood. In this paper, we focus on the coupling mechanism of multi-force interactions in the myosin molecule during its interaction with actin. These forces include the electrostatic force, the van der Waals force and the Casimir force in molecular dynamic simulations of the molecules in solvent with thermal fluctuations. Based on the Hamaker approach, van der Waals and Casimir potentials and forces are calculated between myosin and actin. We have developed a Monte Carlo method to simulate the dynamic activity of the molecular motor. We have shown that because of the retardation effect, the van der Waals force falls into the Casimir force when the distance between the surfaces is larger than 3 nm. When the distance is smaller than 3 nm, the electrostatic force and the van der Waals force increase until the myosin becomes attached to the actin. Over the distances studied in the present work, the electrostatic force dominates the attractive interactions. Our calculations are in good agreement with recently reported experimental results.     

MSAP enhances migration of C6 glioma cells through phosphorylation of the myosin regulatory light chain

A key regulatory mechanism in cell motility is the control of myosin activity, which in non-muscle cells is determined by phosphorylation of the myosin regulatory light chain (MRLC). Here we show that MRLC-interacting protein (MIR)-interacting saposin-like protein (MSAP) enhances cell spreading in fibroblasts and migration of rat C6 glioma cells through increases in MRLC phosphorylation. Overexpression of MSAP enhanced the motility of glioma cells measured in matrigel invasion chambers and using a scratch assay. Downregulation of MSAP by RNA interference significantly decreased glioma cell migration and phosphorylation of MRLC. Inhibition of the corresponding MRLC kinase by ML-7 did not affect migration of MSAP-overexpressing cells. The present results show that MSAP controls glioma cell migration via enhancement of MRLC phosphorylation. This effect is independent of the activity of MRLC kinase. Thus, MSAP is a novel modulator of cell motility that influences migration of glioma cells and possibly other tumors.Received 9 February 2005; received after revision 2 March 2005; accepted 21 March 2005     

抑制肌球蛋白Ⅱ对NRK细胞分裂的影响

研究肌球蛋白Ⅱ在NRK细胞分裂中的作用。分别于分裂中期和后期施加肌球蛋白Ⅱ抑制剂,对细胞分裂进行动态图像采集,采用显微图像分析软件对细胞间桥形态学进行测量分析,采用细胞免疫荧光技术检测肌动蛋白的分布,研究表明:抑制肌球蛋白Ⅱ可引起NRK细胞核分裂受阻或延迟,使子细胞丧失极性,但不影响胞质分裂进程;在胞质分裂早期,抑制肌球蛋白Ⅱ使分裂沟相对直径迅速变细,曲线明显变陡,胞质分裂晚期各组无明显差别。肌球蛋白Ⅱ抑制组与对照组分裂沟处均有肌动蛋白分布,提示肌球蛋白Ⅱ参与了细胞核分裂和子细胞极性的维持,但不影响肌动蛋白在分裂沟定位。在肌球蛋白Ⅱ受抑制的情况下,NRK细胞胞质分裂模式发生了改变。     

肌肉运动的动力学模型

在SIRM（stochastic inclined rods model）模型的基础上，建立一个与时间、空间、温度等因素有关的符合肌肉运动的基本生理过程的理论模型，从微观相互作用的角度为研究肌肉的运动，并给出运动方程的数值解。     

Purification and characterization of myosin from wheat mitochondria

Myosin was purified from wheat mitochondria using DE-52 anion exchange chromatography and Sephacryl S-300 gel filtration. The molecular weight of its heavy chain is about 210 ku, similar to that of muscle myosin Ⅱ(205 ku), and it could be recognized by the polyclonal antibodies against human skeletal muscle myosin Ⅱ. The ATPase activity of the mitochondrial myosin stimulated by F-actin from chicken muscle is 202.5 nmoles Pi/min·mg. The mitochondrial myosin could be activated by Ca²⁺ and was not inhibited by Ca²⁺ at high concentration. The results demonstrate that the myosin of wheat mitochondria shares some similarities with the skeletal muscle myosin Ⅱ.     

Effect of internal deletion of Myosin II on growth and development ofDictyostelium discoideum

Four deletion mutantDictyostelium myosin II heavy chain genes, MyΔ824-941 (Δ1/ 3S2), MyΔ934-1454 (ΔS2), MyΔ934-1194 (ΔS2-1) and MyΔ1 157–1454 (ΔS2-2), were transformed by standard electfoporation into mhcA-cells (T-null), a mutantDictyostelium cell devoid of endogenous myosin II heavy chain gene. The growth, development and formation of fruiting bodies of cells expressing those mutant myosin II s under suspension culture were investigated by comparison with the wild type cell. The results indicate that internal deletion of myosin II affeds the growth and development ofDictyastelium. Furthermore, the longer the length of deletion, the more serious the defect in phenotype.