Geometric trade-off between contractile force and viscous drag determines the actomyosin-based motility of a cell-sized droplet

動きまわる人工細胞、その鍵は摩擦にあり --細胞が狭い空間を利用して運動する仕組みを解明--. 京都大学プレスリリース. 2022-07-21.Cell migration in confined environments is fundamental for diverse biological processes from cancer invasion to leukocyte trafficking. The cell body is propelled by the contractile force of actomyosin networks transmitted from the cell membrane to the external substrates. However, physical determinants of actomyosin-based migration capacity in confined environments are not fully understood. Here, we develop an in vitro migratory cell model, where cytoplasmic actomyosin networks are encapsulated into droplets surrounded by a lipid monolayer membrane. We find that the droplet can move when the actomyosin networks are bound to the membrane, in which the physical interaction between the contracting actomyosin networks and the membrane generates a propulsive force. The droplet moves faster when it has a larger contact area with the substrates, while narrower confinement reduces the migration speed. By combining experimental observations and active gel theory, we propose a mechanism where the balance between sliding friction force, which is a reaction force of the contractile force, and viscous drag determines the migration speed, providing a physical basis of actomyosin-based motility in confined environments

Examining the assembly pathways and active microtubule mechanics underlying spindle self-organization

The bipolar organization of the microtubule-based mitotic spindle is essential for the faithful segregation of chromosomes in cell division. Despite our extensive knowledge of genes and proteins, the physical mechanism of how the ensemble of microtubules can assemble into a proper bipolar shape remains elusive. Here, we study the pathways of spindle self-organization using cell-free Xenopus egg extracts and computer-based automated shape analysis. Our microscopy assay allows us to simultaneously record the growth of hundreds of spindles in the bulk cytoplasm and systematically analyze the shape of each structure over the course of self-organization. We find that spindles that are maturing into a bipolar shape take a route that is distinct from those ending up with faulty structures, such as those of a tripolar shape. Moreover, matured structures are highly stable with little occasions of transformation between different shape phenotypes. Visualizing the movement of microtubules further reveals a fraction of microtubules that assemble between extra poles and push the poles apart, suggesting the presence of active extensile force that prevents pole coalescence. Together, we propose that a proper control over the magnitude and location of the extensile, pole-pushing force is crucial for establishing spindle bipolarity while preventing multipolarity.Comment: 22 pages, 5 + 2 figure

Locally and Globally Coupled Oscillators in Muscle

At an intermediate activation level, striated muscle exhibits autonomous oscillations called SPOC, in which the basic contractile units, sarcomeres, oscillate in length, and various oscillatory patterns such as traveling waves and their disrupted forms appear in a myofibril. Here we show that these patterns are reproduced by mechanically connecting in series the unit model that explains characteristics of SPOC at the single-sarcomere level. We further reduce the connected model to phase equations, revealing that the combination of local and global couplings is crucial to the emergence of these patterns

Nanoscopic changes in the lattice structure of striated muscle sarcomeres involved in the mechanism of spontaneous oscillatory contraction (SPOC)

Muscles perform a wide range of motile functions in animals. Among various types are skeletal and cardiac muscles, which exhibit a steady auto-oscillation of force and length when they are activated at an intermediate level of contraction. This phenomenon, termed spontaneous oscillatory contraction or SPOC, occurs devoid of cell membranes and at fixed concentrations of chemical substances, and is thus the property of the contractile system per se. We have previously developed a theoretical model of SPOC and proposed that the oscillation emerges from a dynamic force balance along both the longitudinal and lateral axes of sarcomeres, the contractile units of the striated muscle. Here, we experimentally tested this hypothesis by developing an imaging-based analysis that facilitates detection of the structural changes of single sarcomeres at unprecedented spatial resolution. We found that the sarcomere width oscillates anti-phase with the sarcomere length in SPOC. We also found that the oscillatory dynamics can be altered by osmotic compression of the myofilament lattice structure of sarcomeres, but they are unchanged by a proteolytic digestion of titin/connectin – the spring-like protein that provides passive elasticity to sarcomeres. Our data thus reveal the three-dimensional mechanical dynamics of oscillating sarcomeres and suggest a structural requirement of steady auto-oscillation

Molecular motors as an auto-oscillator

The organization of biomotile systems possesses structural and functional hierarchy, building up from single molecules via protein assemblies and cells further up to an organ. A typical example is the hierarchy of cardiac muscle, on the top of which is the heart. The heartbeat is supported by the rhythmic contraction of the muscle cells that is controlled by the Ca2+ oscillation triggered by periodic electrical excitation of pacemaker cells. Thus, it is usually believed that the heartbeat is governed by the control system based on a sequential one-way chain with the electrical∕chemical information transfer from the upper to the lower level of hierarchy. On the other hand, it has been known for many years that the contractile system of muscle, i.e., skinned muscle fibers and myofibrils, itself possesses the auto-oscillatory properties even in the constant chemical environment. A recent paper [Plaçais, et al. (2009), Phys. Rev. Lett. 103, 158102] demonstrated the auto-oscillatory movement∕tension development in an in vitro motility assay composed of a single actin filament and randomly distributed myosin II molecules, suggesting that the auto-oscillatory properties are inherent to the contractile proteins. Here we discuss how the molecular motors may acquire the higher-ordered auto-oscillatory properties while stepping up the staircase of hierarchy

Designing Robots as an Humanitarian Project : Implications for Implementing the Course of Study Revised by the MEXT in 2008

設計による問題解決を重視したロボット設計製作題材の有効性に対する認識が定着しつつある。中でも介護や災害時の人命救助を扱う題材は, 技術だけでなく人への優しさなどそのあり方まで考えさせる点で優れている。 一方学習指導要領改訂による必修領域増加の対応策としてロボット題材の検討が求められている。しかし, 1)問題解決の手段としての設計が必要, 2)時間割内で完成させ達成感を得させる必要, 3)創造的設計に挑戦した場合, 完成直前に修正不能であることが判明する場合もあり, 初期段階で実現可能性に気づかせる支援が必要, 4)思いつきの段階から実装動作までの過程で直面する様々な問題について, 力を合わせて解決を図る支援が必要, など多くの課題が存在する。 試行実践により次のような結果を得た。a)構想図に名称や説明の記入により, 言語による共同作業が円滑化した。b)3次元位置の制約が強く, その決定や修正支援が必要。c)作品解体やリサイクルを通した学習も可能。e)ルール等による課題難易度の分布調整が学習に影響する。f)救出対象の荷重値と電流計測により効率の考察も可能。g)教師の問いかけにより生徒の問題解決が促進した, このような事例蓄積と共有化が必要