Autonomous assembly and disassembly of gliding molecular robots regulated by a DNA-based molecular controller.

In recent years, there has been a growing interest in engineering dynamic and autonomous systems with robotic functionalities using biomolecules. Specifically, the ability of molecular motors to convert chemical energy to mechanical forces and the programmability of DNA are regarded as promising components for these systems. However, current systems rely on the manual addition of external stimuli, limiting the potential for autonomous molecular systems. Here, we show that DNA-based cascade reactions can act as a molecular controller that drives the autonomous assembly and disassembly of DNA-functionalized microtubules propelled by kinesins. The DNA controller is designed to produce two different DNA strands that program the interaction between the microtubules. The gliding microtubules integrated with the controller autonomously assemble to bundle-like structures and disassemble into discrete filaments without external stimuli, which is observable by fluorescence microscopy. We believe this approach to be a starting point toward more autonomous behavior of motor protein-based multicomponent systems with robotic functionalities.

Role of tubulin C-terminal tail on mechanical properties of microtubule.

Tubulin C-terminal tail (CTT) is a disordered segment extended from each tubulin monomer of αß tubulin heterodimers, the building blocks of microtubules. The tubulin CTT contributes to the cellular function of microtubules such as intracellular transportation by regulating their interaction with other proteins and cell shape regulation by controlling microtubule polymerization dynamics. Although the mechanical integrity of microtubules is crucial for their functions, the role of tubulin CTT on microtubule mechanical properties has remained elusive. In this work, we investigate the role of tubulin CTTs in regulating the mechanical properties of microtubules by estimating the persistence lengths and investigating the buckling behavior of microtubules with and without CTT. We find that microtubules with intact CTTs exhibit twice the rigidity of microtubules lacking tubulin CTTs. Our study will widen the scope of altering microtubule mechanical properties for its application in nano bio-devices and lead to novel therapeutic approaches for neurodegenerative diseases with altered microtubule properties.

Growth of self-integrated atomic quantum wires and junctions of a Mott semiconductor.

Continued advances in quantum technologies rely on producing nanometer-scale wires. Although several state-of-the-art nanolithographic technologies and bottom-up synthesis processes have been used to engineer these wires, critical challenges remain in growing uniform atomic-scale crystalline wires and constructing their network structures. Here, we discover a simple method to fabricate atomic-scale wires with various arrangements, including stripes, X-junctions, Y-junctions, and nanorings. Single-crystalline atomic-scale wires of a Mott insulator, whose bandgap is comparable to those of wide-gap semiconductors, are spontaneously grown on graphite substrates by pulsed-laser deposition. These wires are one unit cell thick and have an exact width of two and four unit cells (1.4 and 2.8 nm) and lengths up to a few micrometers. We show that the nonequilibrium reaction-diffusion processes may play an essential role in atomic pattern formation. Our findings offer a previously unknown perspective on the nonequilibrium self-organization phenomena on an atomic scale, paving a unique way for the quantum architecture of nano-network.

Motion of a swimming droplet under external perturbations: A model-based approach.

Microdroplets driven by the Marangoni effect are known to continue to swim for hours despite their simple composition. This swimming microdroplet changes its motion from straight to curvilinear and further to chaotic as the Péclet number increases. In this study, we investigate the effect of external perturbations on the three-dimensional axis-asymmetric model of a droplet driven by the Marangoni effect. The aim here is to elucidate the contribution of external perturbation to the complex motion of the droplet and the change in its effect according to the droplet size. In this paper, first we provide a detailed explanation on the derivation of the model introduced in our previous work, which is next used to describe the motion of the droplet in the numerical study of the angular response to random perturbations. The numerical simulation of droplet motion with different types of noise indicates that the model does not converge them into a certain type of motion but rather helps to reflect the external perturbations. The obtained results suggest that the types and properties of external perturbation have a considerable effect on the droplet motion.

Simple dynamics underlying the survival behaviors of ciliates.

Ciliates are swimming microorganisms in aquatic environments. Habitats where ciliates accumulate include nutrient-rich solid-liquid interfaces such as pond bottom walls and waterweed surfaces. The ciliates stay near the walls to survive. We investigated the dynamics of the near-wall behavior of ciliates. In experiments, the ciliates were made to slide on a flat wall of glass substrate. When encountering the wall, the wall-side cilia of the cells stop their motion and lose their propelling activity, which indicates that the ciliates have a mechano-sensing system for cilia beating. Based on the experimental results, we hypothesized that the ciliary thrust force that propels the cell body becomes asymmetric, and the asymmetry of the thrust force generates a head-down torque to keep the cell sliding on the wall. To prove this hypothesis, we performed numerical simulations by using a developed hydrodynamic model for swimming ciliates. The model revealed that the loss of cilia activity on the wall side physically induces a sliding motion, and the aspect ratio of the cell body and effective cilium area are critical functions for the sliding behavior on a wall. In addition, we investigated the stability of the sliding motion against an external flow. We found that ciliates slide upstream on a wall. Interestingly, the dynamics of this upstream sliding, called rheotaxis, were also explained by the identical physical conditions for no-flow sliding. Only two simple physical conditions are required to explain the dynamics of ciliate survival behavior. This review article is an extended version of the Japanese article, Fluid Dynamic Model Reveals a Mechano-sensing System Underlying the Behavior of Ciliates, published in SEIBUTSU BUTSURI Vol. 61, p. 16-19 (2021).

Accumulation of Tetrahymena pyriformis on Interfaces.

The behavior of ciliates has been studied for many years through environmental biology and the ethology of microorganisms, and recent hydrodynamic studies of microswimmers have greatly advanced our understanding of the behavioral dynamics at the single-cell level. However, the association between single-cell dynamics captured by microscopic observation and pattern dynamics obtained by macroscopic observation is not always obvious. Hence, to bridge the gap between the two, there is a need for experimental results on swarming dynamics at the mesoscopic scale. In this study, we investigated the spatial population dynamics of the ciliate, Tetrahymena pyriformis, based on quantitative data analysis. We combined the image processing of 3D micrographs and machine learning to obtain the positional data of individual cells of T. pyriformis and examined their statistical properties based on spatio-temporal data. According to the 3D spatial distribution of cells and their temporal evolution, cells accumulated both on the solid wall at the bottom surface and underneath the air-liquid interface at the top. Furthermore, we quantitatively clarified the difference in accumulation levels between the bulk and the interface by creating a simple behavioral model that incorporated quantitative accumulation coefficients in its solution. The accumulation coefficients can be compared under different conditions and between different species.

Near-wall rheotaxis of the ciliate Tetrahymena induced by the kinesthetic sensing of cilia.

To survive in harsh environments, single-celled microorganisms autonomously respond to external stimuli, such as light, heat, and flow. Here, we elucidate the flow response of Tetrahymena, a well-known single-celled freshwater microorganism. Tetrahymena moves upstream against an external flow via a behavior called rheotaxis. While micrometer-sized particles are swept away downstream in a viscous flow, what dynamics underlie the rheotaxis of the ciliate? Our experiments reveal that Tetrahymena slides along walls during upstream movement, which indicates that the cells receive rotational torque from shear flow to control cell orientation. To evaluate the effects of the shear torque and propelling speed, we perform a numerical simulation with a hydrodynamic model swimmer adopting cilia dynamics in a shear flow. The swimmer orientations converge to an upstream alignment, and the swimmer slides upstream along a boundary wall. The results suggest that Tetrahymena automatically responds to shear flow by performing rheotaxis using cilia-stalling mechanics.

Straight-to-Curvilinear Motion Transition of a Swimming Droplet Caused by the Susceptibility to Fluctuations.

In this Letter, a water-in-oil swimming droplet's transition from straight to curvilinear motion is investigated experimentally and theoretically. An analysis of the experimental results and the model reveal that the motion transition depends on the susceptibility of the droplet's direction of movement to external stimuli as a function of environmental parameters such as droplet size. The simplicity of the present experimental system and the model suggests implications for a general class of transitions in self-propelled swimmers.

Swimming droplets in 1D geometries: an active Bretherton problem.

We investigate experimentally the behavior of self-propelled water-in-oil droplets, confined in capillaries of different square and circular cross-sections. The droplet's activity comes from the formation of swollen micelles at its interface. In straight capillaries the velocity of the droplet decreases with increasing confinement. However, at very high confinement, the velocity converges toward a non-zero value, so that even very long droplets swim. Stretched circular capillaries are used to explore even higher confinement. The lubrication layer around the droplet then takes a non-uniform thickness which constitutes a significant difference to usual flow-driven passive droplets. A neck forms at the rear of the droplet, deepens with increasing confinement, and eventually undergoes successive spontaneous splitting events for large enough confinement. Such observations stress the critical role of the activity of the droplet interface in the droplet's behavior under confinement. We then propose an analytical formulation by integrating the interface activity and the swollen micelle transport problem into the classical Bretherton approach. The model accounts for the convergence of the droplet's velocity to a finite value for large confinement, and for the non-classical shape of the lubrication layer. We further discuss on the saturation of the micelle concentration along the interface, which would explain the divergence of the lubrication layer thickness for long enough droplets, eventually leading to spontaneous droplet division.

Force generation by a propagating wave of supramolecular nanofibers.

Dynamic spatiotemporal patterns that arise from out-of-equilibrium biochemical reactions generate forces in living cells. Despite considerable recent efforts, rational design of spatiotemporal patterns in artificial molecular systems remains at an early stage of development. Here, we describe force generation by a propagating wave of supramolecular nanofibers. Inspired by actin dynamics, a reaction network is designed to control the formation and degradation of nanofibers by two chemically orthogonal stimuli. Real-time fluorescent imaging successfully visualizes the propagating wave based on spatiotemporally coupled generation and collapse of nanofibers. Numerical simulation indicates that the concentration gradient of degradation stimulus and the smaller diffusion coefficient of the nanofiber are critical for wave emergence. Moreover, the force (0.005 pN) generated by chemophoresis and/or depletion force of this propagating wave can move nanobeads along the wave direction.

Influence of cellular shape on sliding behavior of ciliates.

Some types of ciliates accumulate on solid/fluid interfaces. This behavior is advantageous to survival in nature due to the presence of sufficient nutrition and stable environments. Recently, the accumulating mechanisms of Tetrahymena pyriformis at the interface were investigated. The synergy of the ellipsoidal shape of the cell body and the mechanosensing feature of the cilia allow for cells to slide on interfaces, and the sliding behavior leads to cell accumulation on the interfaces. Here, to examine the generality of the sliding behavior of ciliates, we characterized the behavior of Paramecium caudatum, which is a commonly studied ciliate. Our experimental and numerical results confirmed that P. caudatum also slid on the solid/fluid interface by using the same mechanism as T. pyriformis. In addition, we evaluated the effects of cellular ellipticity on their behaviors near the wall with a phase diagram produced via numerical simulation.

DSH5, a dihydrosphingosine C4 hydroxylase gene family member, shows spatially restricted expression in rice and is lethal when expressed ectopically.

Dihydrosphingosine C4 hydroxylase (DSH), a diiron-binding membrane enzyme, catalyzes the hydration of dihydrosphingosine and acyl-sphinganine to produce phytosphingosine and phytoceramide, respectively. Rice has two types of DSH homologs: general DSHs, namely DSH1, DSH2 and DSH4, and others that show spatial expression profiles, namely DSH3 and DSH5. The general DSHs exist in many plant species. These DSHs showed similarity in their functions and complemented the yeast sur2D mutation. In contrast, homologs of DSH3 and DSH5 were found only in monocot plants. Phylogenetic analysis placed these DSHs in different clades that are evolutionarily divergent from those of the general DSHs. DSH3 and DSH5 showed low-level expression. DSH5 expression was specifically in vascular bundle tissues. Ectopic expression of DSH5 induced a dwarf phenotype characterized by severe growth inhibition and an increase in the thickness of the leaf body caused by enlargement of bulliform cells in the leaves. However, no significant difference was observed in the amount of sphingolipid species. DSH5 did not complement the yeast sur2D mutation, implying that DSH5 has little effect on sphingolipid metabolism. These findings suggested that DSH3 and DSH5 originated and diverged in monocot plants.

Self-propelled motion switching in nematic liquid crystal droplets in aqueous surfactant solutions.

The self-propelled motions of micron-sized nematic liquid crystal droplets in an aqueous surfactant solution have been studied by tracking individual droplets over long time periods. Switching between self-propelled modes is observed as the droplet size decreases at a nearly constant dissolution rate: from random to helical and then straight motion. The velocity of the droplet decreases with its size for straight and helical motions but is independent of size for random motion. The switching between helical and straight motions is found to be governed by the self-propelled velocity, and is confirmed by experiments at various surfactant concentrations. The helical motion appears along with a shifting of a point defect from the self-propelled direction of the droplet. The critical velocity for this shift of the defect position is found to be related with the Ericksen number, which is defined by the ratio of the viscous and elastic stresses. In a thin cell whose thickness is smaller than that of the initial droplet size, the droplets show more complex trajectories, including "figure-8s" and zigzags. The appearance of those characteristic motions is attributed to autochemotaxis of the droplet.

Simple mechanosense and response of cilia motion reveal the intrinsic habits of ciliates.

An important habit of ciliates, namely, their behavioral preference for walls, is revealed through experiments and hydrodynamic simulations. A simple mechanical response of individual ciliary beating (i.e., the beating is stalled by the cilium contacting a wall) can solely determine the sliding motion of the ciliate along the wall and result in a wall-preferring behavior. Considering ciliate ethology, this mechanosensing system is likely an advantage in the single cell's ability to locate nutrition. In other words, ciliates can skillfully use both the sliding motion to feed on a surface and the traveling motion in bulk water to locate new surfaces according to the single "swimming" mission.

Geometry-driven collective ordering of bacterial vortices.

Controlling the phases of matter is a challenge that spans from condensed materials to biological systems. Here, by imposing a geometric boundary condition, we study the controlled collective motion of Escherichia coli bacteria. A circular microwell isolates a rectified vortex from disordered vortices masked in the bulk. For a doublet of microwells, two vortices emerge but their spinning directions show transition from parallel to anti-parallel. A Vicsek-like model for confined self-propelled particles gives the point where the two spinning patterns occur in equal probability and one geometric quantity governs the transition as seen in experiments. This mechanism shapes rich patterns including chiral configurations in a quadruplet of microwells, thus revealing a design principle of active vortices.

Non-periodic oscillatory deformation of an actomyosin microdroplet encapsulated within a lipid interface.

Active force generation in living organisms, which is mainly involved in actin cytoskeleton and myosin molecular motors, plays a crucial role in various biological processes. Although the contractile properties of actomyosin have been extensively investigated, their dynamic contribution to a deformable membrane remains unclear because of the cellular complexities and the difficulties associated with in vitro reconstitution. Here, by overcoming these experimental difficulties, we demonstrate the dynamic deformation of a reconstituted lipid interface coupled with self-organized structure of contractile actomyosin. Therein, the lipid interface repeatedly oscillates without any remarkable periods. The oscillatory deformation of the interface is caused by the aster-like three-dimensional hierarchical structure of actomyosin inside the droplet, which is revealed that the oscillation occurs stochastically as a Poisson process.

Dynamic clustering of driven colloidal particles on a circular path.

We studied the collective motion of particles forced to move along a circular path in water by utilizing an optical vortex. Their collective motion, including the spontaneous formation of clusters and their dissociation, was observed. The observed temporal patterns depend on the number of particles on the path and the variation of their sizes. The addition of particles with different sizes suppresses the dynamic formation and dissociation of clusters and promotes the formation of specific stationary clusters. These experimental findings are reproduced by numerical simulations that take into account the hydrodynamic interaction between the particles and the radial trapping force confining the particles to the circular path. A transition between stationary and nonstationary clustering of the particles was observed by varying their size ratio in the binary-size systems. Our simulation reveals that the transition can be either continuous or discontinuous depending on the number of different-size particles. This result suggests that the size distribution of particles has a significant effect on the collective behavior of self-propelled particles in viscous fluids.

Molecular behavior of DNA in a cell-sized compartment coated by lipids.

The behavior of long DNA molecules in a cell-sized confined space was investigated. We prepared water-in-oil droplets covered by phospholipids, which mimic the inner space of a cell, following the encapsulation of DNA molecules with unfolded coil and folded globule conformations. Microscopic observation revealed that the adsorption of coiled DNA onto the membrane surface depended on the size of the vesicular space. Globular DNA showed a cell-size-dependent unfolding transition after adsorption on the membrane. Furthermore, when DNA interacted with a two-phase membrane surface, DNA selectively adsorbed on the membrane phase, such as an ordered or disordered phase, depending on its conformation. We discuss the mechanism of these trends by considering the free energy of DNA together with a polyamine in the solution. The free energy of our model was consistent with the present experimental data. The cooperative interaction of DNA and polyamines with a membrane surface leads to the size-dependent behavior of molecular systems in a small space. These findings may contribute to a better understanding of the physical mechanism of molecular events and reactions inside a cell.

Droplet-Shooting and Size-Filtration (DSSF) Method for Synthesis of Cell-Sized Liposomes with Controlled Lipid Compositions.

We report a centrifugal microfluidic method, droplet-shooting and size-filtration (DSSF), for the production of cell-sized liposomes with controlled lipid compositions. This involves the generation of large and small droplets from the tip of a glass capillary and the selective transfer of small droplets through an oil-water interface, thus resulting in the generation of cell-sized liposomes. We demonstrate control of the microdomain formation as well as the formation of asymmetric lipid bilayer liposomes of uniform size by the control of lipid composition. The DSSF method involves simple microfluidics and is easy to use. In addition, only a small volume (0.5-2 µL) of sample solution is required for the formation of hundreds of cell-sized liposomes. We believe that this method can be applied to generate cell-sized liposomes for a wide variety of uses, such as the construction of artificial cell-like systems.