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[Locomotion of Barents Sea ophiuroids].

Locomotion of 5 species of Ophiuroids from the Barents Sea has been described--Ophiura albida, O. robusta, O. sarsi, Ophiacantha bidentata, Ophiopholis aculeata. Representatives from the genus Ophiura move by means of one or two symmertic pairs of arms which perform rhythmic rowing beats. The feet tube do not participate in their locomotion. Locomotion pattern in O. bidentata and especially O. aculeata, which have long flexible arms, is more complex. However, the analysis indicates that they replace themselves by a comparatiely limited set of standard "programmed" movements which may be used in various combinations. In locomotion of these species, the tube feet are involved which provide for the reliable contact of distal ends of the active rays with the supporting surface; due to this contact the animals can move in a straightforward direction.

Animals↗

Some fundamental aspects of the biomechanics of overground versus treadmill locomotion.

The literature shows a wide difference of opinion about the mechanical equality of, or difference between treadmill and overground locomotion. This difference in opinion is often related to the coordinate system which implicitly or explicitly is used. With help of a few theoretical examples of energy calculations this paper shows that the description of treadmill locomotion with respect to a fixed coordinate system can lead to faulty conclusions. It is concluded that as long as the beltspeed is constant a coordinate system should be used which moves with the belt. In such a system no mechanical difference exists in comparison with overground locomotion with respect to a fixed coordinate system. All differences found in locomotion patterns must therefore originate from other than mechanical causes.

Biomechanical Phenomena↗

Energetics and mechanics for partial gravity locomotion.

The role of gravitational acceleration on human locomotion is not clearly understood. It is hypothesized that the mechanics and energetics of locomotion depend upon the prevailing gravity level. A unique human-rated underwater treadmill and an adjustable ballasting harness were used to stimulate partial gravity environments. This study has two research aspects, biomechanics and energetics. Vertical forces which are exerted by subjects on the treadmill-mounted, split-plate force platform show that peak vertical force and stride frequency significantly decrease (p < 0.05) as the gravity level is reduced, while ground contact time is independent of gravity level. A loping gait is employed over a wide range of speeds (approximately 1.5 m/s to approximately 2.3 m/s) suggesting a change in the mechanics for lunar (1/6 G) and Martian (3/8 G) locomotion. As theory predicts, locomotion energy requirements for partial gravity levels are significantly less than at 1 G (p < 0.05).

Adult↗

The ultrastructure of Entamoeba histolytica locomotion.

This quantitative ultrastructural survey of E. histolytica locomotion in Boyden chambers supports the concept that this parasite is capable of random, chemokinetic and chemotactic motility. An E. histolytica committed to chemotaxis will flatten over the filter, accumulate smaller vacuoles at the front of the cell, and will also project pseudopods and its polarized body towards and alongside the chemoattractant axis, respectively. Other cell features such as cell polarization, membrane ruffling, hyaline, total number of pseudopods and caudal displacement of the nucleus appear to be associated with the locomotion efforts as such, perhaps reflecting speed (chemokinesis) but irrespective of orientation (chemotaxis). Finally, only one of the 11 features that were analyzed (i.e., number of vacuoles) failed to be distinctly associated with any of the movement forms studied. E. histolytica appears to possess the full repertoire of locomotion modalities observed in free moving eukaryots, and its motility translates into ultrastructural landmarks that could be useful indicators of subcellular events related to locomotion.

Animals↗

Learning in the development of infant locomotion.

Infants master crawling and walking in an environment filled with varied and unfamiliar surfaces. At the same time, infants' bodies and skills continually change. The changing demands of everyday locomotion require infants to adapt locomotion to the properties of the terrain and to their own physical abilities. This Monograph examines how infants acquire adaptive locomotion in a novel task--going up and down slopes. Infants were tested longitudinally from their first week of crawling until several weeks after they began walking. Everyday locomotor experience played a central role in adaptive responding. Over weeks of crawling, infants' judgments became increasingly accurate, and exploration became increasingly efficient. There was no transfer over the transition from crawling to walking. Instead, infants learned, all over again, how to cope with slopes from an upright position. Findings indicate that learning generalized from everyday experience traveling over flat surfaces at home but that learning was specific to infants' typical method of locomotion and vantage point. Moreover, learning was not the result of simple associations between a particular locomotor response and a particular slope. Rather, infants learned to gauge their abilities on-line as they encountered each hill at the start of the trial. Change in locomotor responses and exploratory movements revealed a process of differentiation and selection spurred by changes in infants' everyday experience, body dimensions, and locomotor proficiency on flat ground.

Child Development↗

[Types of locomotion in Ophiuroidea].

Locomotor movements of two species of ophiura from the Sea of Japan: Ophiura sarsi vadicola Djakonov and Amphipholis kochii Lütken, were filmed. Ophiura sarsi moves by means of two symmetric pairs of arms which perform rhythmical rowing movements, the fifth arm is passive and directed backwards. Animals of this species do not use tube feet in locomotion. In Amphipholis kochii, three main modes of locomotor movements were found. (1) "Breast stroke" in which two symmetric side arms periodically move forwards, and then push forwards the body and three other arms. In this mode, one arm (leading) is directed forwards, while two others backwards. When the side arms move-forwards or backwards, a wave of successive flexions and extensions propagates along the arm segments. (2) "Pushing" by means of the hind arm and "pulling" with the help of the leading arm. (3) Locomotion by means of the tube feet which perform stepping movements. These three modes of locomotion are observed either separately or in various combinations with each other. In (1) and (2) modes of locomotion the tube feet perform also the coupling of the active arms with the ground which is necessary for the animal movement.

Animals↗

Protrusive activity quantitatively determines the rate and direction of cell locomotion.

Locomoting blebbing cells have been used as a model to obtain novel insight into the mechanisms of cell locomotion. We tested the hypothesis that locomotion can be due to progressive one-sided protrusion of cellular volume into pseudopods. The hypothesis is supported by the finding that the rate and direction of locomotion of individual Walker carcinosarcoma cells can be predicted by sequential measurement of protrusive activity. Protrusive activity at the front is closely associated with forward movement of the rear part of the cell. During bleb formation the cell membrane of Walker carcinosarcoma cells is pushed forward faster (1.2-4.1 microns/sec) than known rates of actin elongation.

Animals↗

Integrin involvement in keratocyte locomotion.

Keratocytes are useful in the study of locomotion because they move rapidly (up to 1 micron/second) while maintaining an almost uniform shape, speed and direction. The smooth gliding motion of the keratocyte requires a precise coordination between adhesion, contractility, and retraction. To ask what role integrins play in keratocyte adhesion and locomotion, either RGD peptides or an anti-beta1 integrin mAb that binds to an ectodomain epitope and inhibits adhesion formation was added to the culture media of moving keratocytes. The response to these reagents depended on three interrelated factors: the dose of RGD/mAb, the apparent adhesion strength of the keratocyte to the substratum and the cell speed. High doses cause keratocytes to quickly and irreversibly round up. At intermediate RGD/mAb doses, keratocytes reestablish adhesion after treatment and briefly resume locomotion until partial detachment recurs. At the lowest doses, disruption of beta1 integrin-mediated adhesion formation destabilizes the lamella, temporarily preventing lamellar extension and forward movement of the cell. With increasing culture time, there is an increase in apparent adhesion and a corresponding marked decrease in locomotory velocity. Under these conditions, high doses of RGD/mAb do not cause keratocytes to detach or even produce detectable lamellar instabilities. We postulate that RGD/mAb competitively inhibits new beta1 integrin mediated adhesion formation that is required to support the rates of lamellar extension necessary for rapid locomotion.

Animals↗

Effects of intraaccumbens administration of SCH-23390 on cocaine-induced locomotion and conditioned place preference.

The effects of systemic (0-1.0 mg/kg) or intraaccumbens (0-1.0 microg/side) administration of SCH-23390 on cocaine-induced (0 or 4.2 mg/kg, i.v.) locomotion, sniffing, and conditioned place preference (CPP) were investigated in rats. After behavioral testing was completed, animals were injected with their respective dose of SCH-23390 into the nucleus accumbens (NAc), followed by a systemic injection of the irreversible antagonist N-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline (EEDQ). Receptors occupied by intraaccumbens SCH-23390, and therefore protected from EEDQ-induced inactivation, were then quantified from autoradiograms of sections labeled with 3H-SCH-23390. Systemic administration of 0.5 and 1.0 mg/kg SCH-23390 reversed cocaine-induced locomotion, sniffing, and CPP, suggesting that stimulation of D1-like receptors is necessary for these behavioral changes. Intraaccumbens administration of 1.0 microg/side SCH-23390 reversed cocaine-CPP, and this dose occupied D1-like receptors primarily in the rostral pole of the NAc. Intraaccumbens administration of 0.5 microg/side SCH-23390 reversed cocaine-induced locomotion. However, this dose occupied a similar number of D1-like receptors in the NAc as a lower and behaviorally ineffective dose of 0.1 microg/side, but occupied more receptors in the caudate-putamen relative to both the 0.1 and 1.0 microg/side doses. These findings suggest that stimulation of D1-like receptors in the NAc is necessary for cocaine-CPP, but not for cocaine-induced locomotion.

Animals↗

The Rho/Rho-kinase and the phosphatidylinositol 3-kinase pathways are essential for spontaneous locomotion of Walker 256 carcinosarcoma cells.

Signal transduction pathways controlling spontaneous locomotion of Walker carcinosarcoma cells are not well understood. We have therefore investigated the role of signalling proteins in development of polarity and locomotion of these cells. Treatment of the cells with 100 ng/ml pertussis toxin had no significant effect on the percentage of polarized cells. In contrast, 2 different phosphatidylinositol 3-kinase inhibitors (wortmannin and LY-294002) markedly reduced the proportion of polarized cells. Spontaneous locomotion of the cells was also significantly inhibited by these two inhibitors. In agreement with these data, we observed localization of the p85alpha subunit of phosphatidylinositol 3-kinase predominantly in the membrane fraction of Walker carcinosarcoma cells, indicating constitutive activation of this enzyme. We also investigated a role of Rho family proteins. Spontaneous development of polarity was almost completely suppressed by electroporation of the cells in the presence of 4 microg/ml C(3) exoenzyme, which specifically ADP-ribosylates and inactivates Rho. Two downstream targets of Rho, the Rho-activated kinases I and II, were also detected predominantly in the particulate membrane fraction, suggesting constitutive activation. A specific Rho-kinase inhibitor (Y-27632) blocked spontaneous polarization and migration in a concentration-dependent manner. Our results indicate that constitutive activation of the Rho/Rho-kinase pathway and of phosphatidylinositol 3-kinase plays an essential part in spontaneous development of polarity and cell locomotion of these cells.

Androstadienes↗

Ionic control of locomotion and shape of epithelial cells: I. Role of calcium influx.

The role of calcium in the induction of locomotion, control of direction of locomotion, and modulation of shape of epithelial cells derived from Xenopus laevis tadpole epidermis is investigated. Local influx of calcium is achieved by electrophoretic release of small amounts of calcium from a micropipette (tip diameter 0.1-0.5 micron) closely apposed to the cell body or lamella. The cells are made permeable for calcium by calcium ionophore A23187, and they are kept in Ca++-free, Mg++-rich EGTA Ringer. Another method used to induce Ca++ influx is local application of A23187 while cells move in normal culture medium. Influx of Ca++ into the lamella induces a localised increase in thickness and enlargement of the lamella. Stationary cells become active and show movement in the direction of the Ca++ gradient. Fried-egg-shaped cells tend to acquire a semicircular shape and start moving. Moving cells change the direction of their locomotion, following the direction of Ca++ release. Influx of Ca++ in the cell body region induces its contraction concomitant with an increase in lamellar area. These observations suggest the presence of two different Ca++-sensitive components: an actomyosin meshwork in the cell body and an actin gel in the lamella. Influx of Ca++ induces contraction of actomyosin and solation of actin gel. Interaction of these two systems would explain modulation of shape and generation of locomotion in epithelial cells.

Animals↗

Ionic control of locomotion and shape of epithelial cells: II. Role of monovalent cations.

The migration of keratocytes isolated from Xenopus tadpole epidermis has been investigated in vitro. In saline the cells move with a mean speed of 5-6 microns/min. Migration is slowed down in saline with diminished sodium content and ceases in media containing not more than 4 mM sodium. Inhibition of the Na+/K+-2Cl- cotransporter by piretanide reduces the speed of migrating cells to about one-third of the control level, the same accounts to inhibition of the Na+/H+ antiport with amiloride at pH 7.2. At pH 6.6, however, amiloride only slightly influences locomotion. Depolarization of the plasma membrane by increased extracellular K+ concentration or by inhibition of the Na+/K+ pump by ouabain is only of minor influence during more than 1 h. Hyperpolarization of the cells using the sodium ionophore monensin impedes locomotion; this inhibition depends on an active Na+/K+ pump. Ionophore-mediated breakdown of the K+ gradient strictly inhibits locomotion. The experiments have shown that a continuous flux of sodium ions is indispensable for the maintenance of cell locomotion. These ions may exert their action primarily by affecting cytosolic free calcium concentration and pH.

Amiloride↗

Topographic compensation: guidance and directed locomotion of fibroblasts on grooved micromachined substrata in the absence of microtubules.

Fibroblasts cultured on grooved substrata align themselves and migrate in the direction of the grooves, a phenomenon called contact guidance. Microtubules have been deemed important for cell polarization, directed locomotion, and contact guidance. Because microtubules were the first cytoskeletal element to align with the grooves when fibroblasts spread on grooved substrata, we investigated the consequences of eliminating the influence of microtubules by seeding fibroblasts onto smooth and grooved micromachined substrata in the presence of colcemid. Fibroblasts were examined by time-lapse cinematography and epifluorescence or confocal microscopy to determine cell shape and orientation and the distribution of cytoskeletal or associated elements including actin filaments, vinculin, intermediate filaments, microtubules, and kinesin. As expected, cells spreading on smooth surfaces in the presence of colcemid did not polarize or locomote. Surprisingly however, by 24 hours, cells spread on grooves in the presence of colcemid were morphologically indistinguishable from controls spread on grooves. Both groups were aligned and polarized with the direction of the grooves and demonstrated directional locomotion along the grooves. In the absence of microtubules, kinesin localized to some of the aligned stress fibers and to leading edges of cells spreading on grooves. The grooved substratum compensated for the microtubule deficiency by organizing and maintaining an aligned actin filament framework. Thus, microtubules are not required to establish or maintain stable, polarized cell shapes or directed locomotion, provided an alternate oriented cytoskeletal component is available.

Actins↗

Traction forces in locomoting cells.

A means of determining quantitative maps of the tractions exerted by locomoting cells on a substratum has been developed. This method is similar to the Harris silicone substratum assay [Harris et al., 1980: Science 208:177-179], but uses an improved non-wrinkling film that deforms more predictably in response to traction forces. The method also utilizes a mathematical analysis of rubber deformation to produce the final map of the distribution of tractions. The resulting maps consistently showed that fish keratocytes exert a steady-state "pinching" on the substratum, perpendicular to the cell's direction of locomotion. No significant rearward tractions were detected at or near the front edge of the cell. Likewise, no significant forward tractions associated with peeling of adhesions were found at the back of the cell. A second assay uses deflection of a lightly attached glass microneedle to measure the total force exerted by locomoting cells. Forces of approximately 4.5 x 10(-3) dyn were required to "stall" locomoting keratocytes. The implications of these findings for cell movement are discussed.

Animals↗

Identification of a novel heparin binding domain in RHAMM and evidence that it modifies HA mediated locomotion of ras-transformed cells.

We have previously reported that the hyaluronan (HA) receptor RHAMM (Receptor for HA Mediated Motility) [Turley et al., 1991] contains two HA binding motifs located within a 35 amino acid region of its C-terminus end [Yang et al., 1993] and that HA stimulation of the motility of ras-transformed fibroblasts is mediated via its interaction with RHAMM. Here we show that RHAMM also contains binding sites for heparin (HP) and that interaction of HP with these sites can regulate the locomotion of ras-transformed fibroblasts. At low concentrations (0.01 mg/ml), HP inhibited HA-induced locomotion of ras-transformed cells in a manner independent of RHAMM. At higher, but still physiological concentrations (0.1 mg/ml), HP alone stimulated cell locomotion and this stimulation appeared to be RHAMM-dependent as it was blocked by anti-RHAMM antibodies. Other related glycosaminoglycans such as chondroitin sulfate and dermatin sulfate had no effect on cell motility. In ligand blotting assays, GST-RHAMM fusion protein was shown to bind biotin-labelled HP and this binding was displaceable with unlabelled HP. In similar ligand binding analyses conducted with truncations of RHAMM fusion protein, the HP binding region was found to be localized in the same 35 amino acid segment of RHAMM that contains the two HA binding domains. Synthetic peptides corresponding to these HA binding domains were retained on and bound effectively to an HP-Sepharose affinity column. Fusion proteins generated by linkage of these peptides to the non-HP binding amino terminus of RHAMM conferred HP binding capacity to the genetically engineered proteins. Conversely, deletion of the HA binding domains of RHAMM resulted in fusion proteins devoid of HP binding activity. The relative affinities of RHAMM for HA and HP, as determined by competition and transblot assays as well as quantification of binding at various salt concentrations, indicated that RHAMM had lower affinity for HP than that for HA. These results demonstrate the existence of a new HP binding motif that has biological relevance to cell locomotion.

Amino Acid Sequence↗

Ruffling and locomotion: role in cell resistance to growth factor-induced proliferation.

It has long been known that the growth rate of cells in vitro can be retarded by providing substrates of restricted area. Such experiments were performed with adhesive islets, made by depositing metals onto agarose layers through templates of various sizes. Since normal cells are unable to adhere to agarose, they become confined to the metallic surface. Using such haptotactic islets, we have studied the role of membrane ruffling and cell locomotion in the resistance of AG1523 human fibroblasts to growth factor-induced mitogenesis. Cells plated on small substrates, i.e., 2,150 microns 2 in area, initially showed vigorous ruffling, which was suppressed by 8 h after plating but had resumed again by 12 h. In contrast, cells on larger-size islets showed a rapid decline and stabilization of ruffling activity. When the growth rate was measured for single cells cultured on haptotactic islets, it was found to increase linearly from areas of 4,280 microns 2 up to 425,000 microns 2. Since the area needed to saturate the growth response was approximately 50-fold larger than the area occupied by a single cell, the growth inhibition was attributed in part to an interference with locomotion. The implication that locomotion provided positive input into growth control mechanisms was subjected to a direct test by evaluating the effect of nine polypeptide growth factors on the motility of serum-starved cells. All except TGF-beta 1 stimulated movement. Finally, the mitogenic effect of growth factors was measured by [3H]thymidine incorporation and found to be proportional to motile activities, as quantitatively assayed. We conclude that locomotion suppression is a factor in AG1523 cell resistance to growth factor-induced mitogenesis.

Cell Division↗

Dose and behavioral context dependent inhibition of movement and basal ganglia neural activity by Delta-9-tetrahydrocannabinol during spontaneous and treadmill locomotion tasks in rats.

The effects of Delta-9-tetrahydrocannabinole (Delta-9-THC) on locomotor activities and related basal ganglia neural responses were investigated in rats. A multiple-channel, single unit recording method was used to record neuronal activity in the dorsal lateral striatum, the globus pallidus, the subthalamic nucleus, and the substantia nigra pars reticulata simultaneously during spontaneous movement and treadmill locomotion. Delta-9-THC treatment (0.05-2.0 mg/kg, i.p.) dose-dependently decreased spontaneous motor activity and altered walking patterns in treadmill locomotion in that stance time was increased and step number was decreased. In parallel with the behavioral effects, Delta-9-THC treatment inhibited neural activity across all four basal ganglia areas recorded during both motor tests. Further, this inhibition of basal ganglia neural activity was behavioral context-dependent. Greater inhibition was found during resting than during walking periods in the treadmill locomotion test. Delta-9-THC treatment also changed firing patterns in the striatum and globus pallidus. More neurons in these regions discharged in an oscillatory pattern during treadmill walking with Delta-9-THC, and the oscillatory frequency was similar to that of the step cycle. Synchronized firing patterns were found in few basal ganglia neurons in the control condition (approximately 1%). Synchronized firing patterns increased during the treadmill resting phase after Delta-9-THC treatment, but still represented a very small proportion of the total neural population (1.9%). The drug treatment did not change neural responses to the tone cue proceeding treadmill locomotion. This study demonstrates dose-dependent inhibitory effects of cannabinoid injection on motor activity. This effect may be related to the behavioral context-dependent inhibition observed in the basal ganglia system where CB1 receptors are densely distributed.

Action Potentials↗

T lymphocytes and neutrophil granulocytes differ in regulatory signaling and migratory dynamics with regard to spontaneous locomotion and chemotaxis.

Chemotactic migration of T lymphocytes and neutrophil granulocytes within a three-dimensional collagen matrix is distinct from spontaneous, matrix-induced migration concerning dynamic parameters and regulatory intracellular signaling. Both spontaneous T lymphocyte locomotion and stromal-cell-derived factor-1 (SDF-1)-induced chemotaxis-involved protein tyrosine kinase (PTK) activity, whereas only SDF-1-induced migration was protein kinase C (PKC) dependent. Spontaneous locomotion of neutrophil granulocytes was independent of PKC and PTK activity, but formyl-methionyl-leucyl-phenylalanine-induced migration involved PKC activity. In addition, the microtubule cytoskeleton was not changed after induction of chemotaxis in both cell types. T lymphocytes had a well-developed microtubule cytoskeleton with the microtubule organizing center located in the uropod, whereas neutrophil granulocytes revealed a clustered tubulin distribution at the leading edge of the migrating cell. Therefore, differences of the microtubule cytoskeleton might contribute to differences in locomotion between T lymphocytes and neutrophil granulocytes but not to differences between spontaneous locomotion and chemotaxis.

Cell Movement↗