The development of locomotor kinematics in neonatal rats: an agent-based modeling analysis in group and individual contexts

An agent-based model of infant rat (pup) locomotion and aggregation was developed by modifying a previous model of pup aggregation [Schank, J.C., Alberts, J.R., 2000a. The developmental emergence of coupled activity as cooperative aggregation in rat pups. Proc. R. Soc. London B 267, 2307-2315]. The main difference between the earlier and current models is the incorporation of whole-body kinematics of directional locomotion. Data on locomotion and aggregation are presented for individuals and groups of 7- and 10-day-old pups and the data were used to evolve models (with a genetic algorithm) that fit these data. Aggregation between 7- and 10-day-old pups was considerably different and could be explained by agent-based models, in particular, models with directional-kinematic matrices specifying the probabilities of moving to adjacent cells. The directional kinematics of whole-body movement differed between the two age classes and differed between group and individual contexts for 10-day-old pups. This may indicate a developmental transition (by day 10) to more central control of behavior and the ability to change patterns of movement based on social context. The behavior analyzed with agent-based models may provide a precise way to measure motor and nervous system development in rats and other rodents.     

Velocity distribution of the anterograde and retrograde transport of extracellular particles byAmoeba proteus

Summary The transverse velocity profiles of the anterograde flow of particles on the cell surface and around it are approximately parabolic. The peak velocity is recorded close to the membrane and the descendent arm of the profile is viscosity-dependent. It indicates that the extracellular forward flow is probably generated by a forward movement of the fluid fraction of the membrane itself. The retrograde component of extracellular movements is manifested by particles kept on the cell surface by adhesion, which behave exactly as the ectoplasmic layer on the opposite side of the membrane,i.e., they probably reflect the movement of that fraction of the surface material which is attached to the cortical microfilaments. In the longitudinal profile, the velocity of anterograde flow rises from the tail to the front of amoeba, but is generally related to the effective cell locomotion rate and not to the movements of any intracellular layer. Around the cells deprived of any attachment to the substratum, which cannot locomote but manifest vigorous intracellular movements, the anterograde flow ceases at least along 2/3 of their lenght. It persists, however, around the frontal fountain zone, where other particles still move backwards together with the retracted ectoplasmic layer. This indicates that the role of the forward flow of and on the cell surface is to compensate for: (1) the increase of the surface area in the frontal regions due to locomotion, (2) the withdrawal of a part of material which is hauled back by the retracting cortical layer. A comprehensive scheme of the velocity distribution within the different layers of a moving amoeba and around it has been constructed on the basis of present and earlier data.Study supported by the Research Project CPBP 04.01 of the Polish Academy of Science.I dedicate this paper to Professor K. E. Wohlfarth-Bottermann with the best wishes for his 65th birthday.     

Locomotion ofAmoeba proteus after standardizing its body shape

Summary Amoeba proteus obliged to follow dark stripes in the form of Y may be studied in three repeatable simple configurations: 1. tail + 1 advancing front, 2. tail + 2 advancing pseudopodia, 3. tail +1 advancing pseudopodium + 1 contracting pseudopodium. Formation of two advancing pseudopodia and the later conversion of one of them into a contracting pseudopodium affect the rate of movement of all the other body parts in the manner predictible by the hydrodynamic concept of the endoplasmic flow in amoeba. An active front stops and begins to retreat when arriving to a constant distance from the posterior body end. The locomotion is disfavoured if new pseudopodia deviate from the former body axis at the angle wider than 35°.Study supported by Research Project II. 1 of the Polish Academy of Science.     

Reconstruction of Active Regular Motion in Amoeba Extract: Dynamic Cooperation between Sol and Gel States

Amoeboid locomotion is one of the typical modes of biological cell migration. Cytoplasmic sol–gel conversion of an actomyosin system is thought to play an important role in locomotion. However, the mechanisms underlying sol–gel conversion, including trigger, signal, and regulating factors, remain unclear. We developed a novel model system in which an actomyosin fraction moves like an amoeba in a cytoplasmic extract. Rheological study of this model system revealed that the actomyosin fraction exhibits shear banding: the sol–gel state of actomyosin can be regulated by shear rate or mechanical force. Furthermore, study of the living cell indicated that the shear-banding property also causes sol–gel conversion with the same order of magnitude as that of shear rate. Our results suggest that the inherent sol–gel transition property plays an essential role in the self-regulation of autonomous translational motion in amoeba.     

Depolarization of the cell membrane causes inhibition of cell locomotion and pinocytosis inAcanthamoeba castellanii andAmoeba proteus

Summary 10 M CCCP protonophore in an acidic medium causes depolarization of the cell membrane and immediate cessation of locomotion inAcanthamoeba castellanii andAmoeba proteus. In the basic media there is no depolarization or inhibition of cell locomotion. Other depolarizing agents (alkali cations, crown molecules) also stop locomotion and induce pinocytosis in amoeba. Pinocytotic uptake of horseradish peroxidase byAcanthamoeba castellanii is increased by 69% in the presence of CCCP in the medium at pH 5.7 but is not influenced at higher pH values. This might indicate that both amoeboid locomotion and pinocytosis are controlled by membrane potential.     

Locomotive mechanism of Physarum plasmodia based on spatiotemporal analysis of protoplasmic streaming

We investigate how an amoeba mechanically moves its own center of gravity using the model organism Physarum plasmodium. Time-dependent velocity fields of protoplasmic streaming over the whole plasmodia were measured with a particle image velocimetry program developed for this work. Combining these data with measurements of the simultaneous movements of the plasmodia revealed a simple physical mechanism of locomotion. The shuttle streaming of the protoplasm was not truly symmetric due to the peristalsis-like movements of the plasmodium. This asymmetry meant that the transport capacity of the stream was not equal in both directions, and a net forward displacement of the center of gravity resulted. The generality of this as a mechanism for amoeboid locomotion is discussed.     

Directional persistence of EGF-induced cell migration is associated with stabilization of lamellipodial protrusions

Migrating cells can sustain a relatively constant direction of lamellipodial protrusion and locomotion over timescales ranging from minutes to hours. However, individual waves of lamellipodial extension occur over much shorter characteristic times. Little understanding exists regarding how cells might integrate biophysical processes across these disparate timescales to control the directional persistence of locomotion. We address this issue by examining the effects of epidermal growth factor (EGF) stimulation on long-timescale directional persistence and short-timescale lamellipodial dynamics of EGF receptor-transfected Chinese hamster ovary cells migrating on fibronectin-coated substrata. Addition of EGF increased persistence, with the magnitude of increase correlating with fibronectin coating concentration. Kymographic analysis of EGF-stimulated lamellipodial dynamics revealed that the temporal stability of lamellipodial protrusions similarly increased with fibronectin concentration. A soluble RGD peptide competitor reduced both the persistence of long-timescale cell paths and the stability of short-timescale membrane protrusions, indicating that cell-substratum adhesion concomitantly influences lamellipodial dynamics and directional persistence. These results reveal the importance of adhesion strength in regulating the directional motility of cells and suggest that the short-timescale kinetics of adhesion complex formation may play a key role in modulating directional persistence over much longer timescales.     

Three New Limax Amoebae Isolated from Marine Surface Sediments: Vahlkampfia caledonica N. Sp., Saccamoeba marina N. Sp., and Hartmannella vacuolata N. Sp.

ABSTRACT. Three new limax amoebae, isolated from marine, surface sediment samples are described using light microscopic and fine structural features. One species, characterized by eruptive locomotion typical of the family Vahlkampfiidae, is assigned the name Vahlkampfia caledonica (47.4 ± 16.0 μm × 12.1 ± 3.2 μm). The other two monopodial species move with steady locomotion characteristic of the family Hartmannellidae. One is a Saccamoeba with a distinct posterior bulbous uriod, vaculoes containing prominent crystals, glycocalyx with cup-like components, and spherical nucleus with central nucleolus. It is assigned the name Saccamoeba marina (72.5 ± 14.9 ±μm × 20. 7 ± 4. 5 μM). The other hartmannellid limax amoeba moves by steady locomotion and has a rather constant monopodial from, lacks a uroid, but has occasional trailing masses of cytoplasm, contains cup-like structures in the glycocalyx, and is characterized μm). Few limax amoebae have been described from marine environment and these data provide additional evidence that limax amoebae may be more abundant in marine sediments that realized previously.     

The Arp2/3 inhibitory protein Arpin is dispensable for chemotaxis

Arpin is an Arp2/3 inhibitory protein, which decreases the protrusion lifetime and hence directional persistence in the migration of diverse cells. Arpin is activated by the small GTPase Rac, which controls cell protrusion, thus closing a negative feedback loop that renders the protrusion intrinsically unstable. Because of these properties, it was proposed that Arpin might play a role in directed migration, where directional persistence has to be fine‐tuned. We report here, however, that Arpin‐depleted tumour cells and Arpin knock‐out Dictyostelium amoeba display no obvious defect in chemotaxis. These results do not rule out a potential role of Arpin in other systems, but argue against a general role of Arpin in chemotaxis.     

Navigating signaling networks: chemotaxis in Dictyostelium discoideum

Studies of chemotaxis in the social amoeba Dictyostelium discoideum have revealed numerous conserved signaling networks that are activated by chemoattractants. In the presence of a uniformly distributed stimulus, these pathways are transiently activated, but in a gradient they are activated persistently and can be localized to either the front or the back of the cell. Recent studies have begun to elucidate how chemoattractant signaling regulates the three main components of chemotaxis: directional sensing, pseudopod extension, and polarization.     

Cell-substrate interactions during amoeboid locomotion of neutrophil leukocytes

Cell-substrate interactions between human blood neutrophils moving on a glass substrate in serum-free medium have been investigated using reflexion interference microscopy, high voltage and scanning electron microscopy (SEM). The contact pattern with the substrate differed considerably from that found in fibroblasts and the amoeba Naegleria. Discrete focal contacts could not be detected but large broad areas of very close contact (accounting for about 30% of the total contact area) could be found particularly associated with the uroid. Considerable loss of membrane material occurred as a result of breakdown of the uroid during locomotion.     

B-cell subsets responsive to fluorescein-conjugated antigens. I. Sensitivity to anti-IgD, tolerance, and cross-priming

Human peripheral blood monocyte-depleted lymphocytes, T lymphocytes, and non-T lyphocytes were studied for their locomotor activity in response to several common chemotactic stimuli. The factors used to stimulate lymphocyte locomotion were casein, C5a, and f-Met-Leu-Phe. Chemotaxis (directional locomotion) as well as chemokinesis (nondirectional locomotion) in response to each factor were delineated. Monocyte-depleted lymphocyte locomotion was stimulated significantly by all of the above factors. Separation of lymphocytes into T cells and non-T cells indicated that T-lymphocyte locomotion was stimulated by casein and C5a but not by f-Met-Leu-Phe. Non-T lymphocytes were found to respond to C5a and f-Met-Leu-Phe but responded minimally to casein. Additional experiments indicated that casein and f-Met-Leu-Phe were chemokinetic for both monocyte-depleted lymphocytes and non-T lymphocytes, while C5a was chemotactic for both monocyte-depleted lymphocyte preparations and purified T cells.     

Cell polarization and adhesion in a motile pathogenic protozoan: role and fate of the Entamoeba histolytica Gal/GalNAc lectin

The human pathogenic protozoan Entamoeba histolytica is a motile cell polarized into a front pseudopod and a rear uroid. The amoebic Gal/GalNAc surface lectin is a major adhesion molecule composed of an immunodominant 170-kDa heavy subunit, mostly extracellular except for a short cytoplasmic tail, and of an extracellular light subunit. The binding of multivalent ligands triggers lectin capping and recruitment to the uroid. The properties of the Gal/GalNAc lectin and its role in amoeba adhesion and uroid polarization are reviewed in the context of the molecular mechanisms underlying cell polarization and locomotion.     

Ultrastructure and Description of a Fungus-Feeding Amoeba,Trichamoeba mycophaga n. sp. (Amoebidae,Amoebea), from Australia1

ABSTRACT. An amoeba isolated from a wheatfield and a forest soil in Australia has been identified as Trichamoeba mycophaga n. sp. Trophozoites of this amoeba are palmate to elongate and measure 45–136 μm in length and 25–94 μm in width. Amoebae in continuous locomotion may be limax with a villous-bulb uroid. Both the lobose pseudopodia and the advancing margin of a limax trophozoite bear an ectoplasmic crescent. The plasma membrane is coated with an electron-dense amorphous layer ca. 100 nm thick. Endoplasm is granular with elongate to bipyramidal crystals and contains bacterial endosymbionts. Trophozoites have a single, spherical to oval nucleus, 4–10 μm in diameter, which contains a centrally located, spherical to oval nucleolus, 2.8–5.0 μm in diameter. The nucleoplasm contains aggregations of filaments distributed radially within the nuclear membrane. Cysts are 21–60 μm in diameter, with ecto- and endocyst walls separated by an amorphous layer.     

An imbalancing act: gap junctions reduce the backward motor circuit activity to bias C. elegans for forward locomotion

A neural network can sustain and switch between different activity patterns to execute multiple behaviors. By monitoring the decision making for directional locomotion through motor circuit calcium imaging in?behaving Caenorhabditis elegans (C.?elegans), we reveal that C.?elegans determines the directionality of movements by establishing an imbalanced output between the forward and backward motor circuits and that it alters directions by switching between these imbalanced states. We further demonstrate that premotor interneurons modulate endogenous motoneuron activity to establish the output imbalance. Specifically, the UNC-7 and UNC-9 innexin-dependent premotor interneuron-motoneuron coupling prevents a balanced output state that leads to movements without directionality. Moreover, they act as shunts to decrease the backward-circuit activity, establishing a persistent bias for the high forward-circuit output state that results in the inherent preference of C.?elegans for forward locomotion. This study demonstrates that imbalanced motoneuron activity underlies directional movement and establishes gap junctions as critical modulators of the properties and outputs of neural circuits.     

Adhesion,Morphology, and Locomotion of Paramoeba pemaquidensis Page (Amoebida,Paramoebidae): Effects of Substrate Charge Density and External Cations1

Morphology and locomotive behavior in the marine amoeba, Paramoeba pemaquidensis Page, was examined under different environmental conditions. Paramoeba requires a minimum surface negative charge density for adhesion of amoebae to substrata. Once adhesion to the substratum has been attained, however, surface negative charge density has no effect on morphology or locomotive rate. Divalent cations are not required for adhesion, but external calcium is required for normal locomotion. In the presence of calcium, Paramoeba often assumes a locomotive form with a broad, well-developed anterior hyaline region and truncate posterior region. Locomotive forms vary from those with only a well-developed hyaline region (Flabellula-like) to forms with long digitiform sub-pseudopodia (Vexillifera-like), with intermediate morphotypes. Locomotive rates decrease and anteroposterior polarity disappears in the presence of living or heat-killed bacteria, indicating that phagocytosis temporarily interferes with locomotion and alters form.     

Impaired neutrophil locomotion associated with hyperadhesiveness in a patient with Chédiak-Higashi syndrome

Neutrophils from a patient with Chédiak-Higashi (CH) syndrome exhibited defective directional locomotion in a gradient of activated plasma. Further analysis of the nature of this defect showed that CH neutrophils could respond normally to stimulation with f-Met-Leu-Phe, by increasing both their motility and polarization, provided the cells were kept in suspension. Contact with the substratum resulted in the loss of both motility and polarity in the majority of cells. CH neutrophils, in contrast to normal cells, did not respond chemokinetically to f-Met-Leu-Phe. Unstimulated random locomotion of CH neutrophils was also depressed, and this correlated with increased spreading on the substratum. Our results indicate that motility, locomotion and polarisation of CH neutrophils on the substratum are depressed because of excessive adhesion.     

Modulation of Biological Functions of Naegleria fowleri Amoebae by Growth Medium

Two strains of Naegleria fowleri amoebae were studied when the amoebae were maintained in the same growth medium or in two different media. A weakly pathogenic strain of N. fowleri , LEE, and a highly pathogenic strain, LEEmpCl, were compared for growth properties, the presence or absence of surface structures termed food cups, cytopathogenicity, cellular locomotion, susceptibility to complement-mediated lysis and immunological relatedness by western immunoblot analysis when grown in Nelson medium or in Cline medium. The two different strains of N. fowleri , LEE and LEEmpCl, were more similar in protein profiles and functional activity when both strains were grown in the same nutritional medium. Differences in growth, proteins synthesized, cytopathogenicity, susceptibility to complement lysis and rate of locomotion were noted when the same strain was grown in different media. Naegleria fowleri grown in Cline medium demonstrated an increased rate of growth, an increase in its rate of locomotion, an increased resistance to complement lysis, and destroyed target nerve cells by contact-dependent lysis. In contrast, the same strain of amoeba grown in Nelson medium showed slower growth, destroyed target cells by trogocytosis, and was less resistant to complement-mediated lysis.     

A visual analysis of chemotactic and chemokinetic locomotion of human neutrophil leucocytes. Use of a new chemotaxis assay with Candida albicans as gradient source.

The locomotion of human blood neutrophil leucocytes was observed and analysed by time-lapse cinematography (1) under conditions where chemokinetic locomotion was stimulated, i.e. in a uniform concentration of casein; (2) in response to chemotactic gradients generated at a point-like source, namely blastospores of the pathogenic yeast Candida albicans in normal human plasma, and (3) in response to soluble chemotactic factors diffusing from Sephadex beads. Neutrophils moving in purely chemokinetic conditions tended to persist in straight paths and showed a preference for narrow angles of turn suggesting a “persistent random walk” type of locomotion rather than a pure random walk. Cells responding to Candida spores showed near straight-line locomotion to the gradient source over short distances (ca 50 μm) and brief time periods. They phagocytosed the spores on arrival and were usually immediately able to respond to a new gradient. Colchicine treatment caused the cells to turn through wider angles, but they were still able to home onto and phagocytose the spores. Colchicine-treated cells showed bizarre and fluctuating shapes but were nonetheless usually polarized towards the gradient source. Gradients from large sources, such as Sephadex beads containing soluble chemotactic factors, were more easily disturbed than those from Candida spores and directional locomotion of cells towards the beads was only seen in certain sectors. The angles of turn made by moving cells under these conditions were an important determinant of chemotaxis since paths of those cells reaching beads showed longer straight segments and narrower angles of turn than those which failed to show a directional response.     

Signaling through the phosphatidylinositol 3-kinase regulates mechanotaxis induced by local low magnetic forces in Entamoeba histolytica

In micro-organisms, as well as in metazoan cells, cellular polarization and directed migration are finely regulated by external stimuli, including mechanical stresses. The mechanisms sustaining the transduction of such external stresses into intracellular biochemical signals remain mainly unknown. Using an external magnetic tip, we generated a magnetic field gradient that allows migration analysis of cells submitted to local low-intensity magnetic forces (50 pN). We applied our system to the amoeba Entamoeba histolytica. Indeed, motility and chemotaxis are key activities that allow this parasite to invade and destroy the human tissues during amoebiasis. The magnetic force was applied either inside the cytoplasm or externally at the rear pole of the amoeba. We observed that the application of an intracellular force did not affect cell polarization and migration, whereas the application of the force at the rear pole of the cell induced a persistent polarization and strongly directional motion, almost directly opposed to the magnetic force. This phenomenon was completely abolished when phosphatidylinositol 3-kinase activity was inhibited by wortmanin. This result demonstrated that the applied mechanical stimulus was transduced and amplified into an intracellular biochemical signal, a process that allows such low-intensity force to strongly modify the migration behavior of the cell.