An analytical solution describing the propagation of positive injury signals in an axon: effect of dynein velocity distribution

A model describing the propagation of positive injury signals from the lesion site in an axon towards the neuron soma is described. It is assumed that these signals are driven by dynein molecular motors. An analytical solution that accounts for the probability density function (pdf) of a dynein velocity distribution is obtained. Two examples of pdf of dynein velocity distributions that follow from the results published in Ross et al. (2006, Processive bidirectional motion of dynein–dynactin complexes in vitro. Nat Cell Biol. 8:562–570) and Deinhardt et al. (2006, Rab5 and Rab7 control endocytic sorting along the axonal retrograde transport pathway. Neuron 52:293–305) are considered. The effect of dynein velocity distribution on the rate of spreading of the signal wave is discussed. It is demonstrated that the obtained solution can be applied to the problem of how neurons measure the distance between the lesion site and the neuron soma.     

Can a death signal half-life be used to sense the distance to a lesion site in axons?

Neuron response to injury depends on the distance to the lesion site, which means that neurons are capable of sensing this distance. Several mechanisms explaining how neurons can do this have been proposed and it is possible that neurons use a combination of several mechanisms to make such measurements. In this paper we investigate the feasibility of the simplest mechanism, which is based on the hypothesis that death signals, produced at the lesion site, propagate toward the neuron soma. The signals are propelled by dynein motors. If signals have a finite half-life, they decay as they propagate. By measuring the concentration of death signals arriving to the soma, neurons should thus be able to determine the distance to the injury site. We develop and solve a transport equation based on the above model. We investigate how a death signal distribution depends on the dynein velocity distribution. We evaluate the efficiency of such a mechanism by investigating the sensitivity of death signal concentration at the soma to the distance to the injury site. By using the hypothesis that system performance is optimized by evolution, we evaluate death signal half-lives that would maximize this sensitivity.     

Effect of fuel concentration and force on collective transport by a team of dynein motors

Motor proteins are essential components of intracellular transport inside eukaryotic cells. These protein molecules use chemical energy obtained from hydrolysis of ATP to produce mechanical forces required for transporting cargos inside cells, from one location to another, in a directed manner. Of these motors, cytoplasmic dynein is structurally more complex than other motor proteins involved in intracellular transport, as it shows force and fuel (ATP) concentration dependent step‐size. Cytoplasmic dynein motors are known to work in a team during cargo transport and force generation. Here, we use a complete Monte‐Carlo model of single dynein constrained by in vitro experiments, which includes the effect of both force and ATP on stepping as well as detachment of motors under force. We then use our complete Monte‐Carlo model of single dynein motor to understand collective cargo transport by a team of dynein motors, such as dependence of cargo travel distance and velocity on applied force and fuel concentration. In our model, cargos pulled by a team of dynein motors do not detach rapidly under higher forces, confirming the experimental observation of longer persistence time of dynein team on microtubule under higher forces.     

The drift distances and time spent in the drift by freshwater shrimps, Gammarus pulex, in a small stony stream, and their implications for the interpretation of downstream dispersal

1. The objectives were (i) to determine experimentally and to model the relationship between mean water velocity and both the mean distance travelled, and the mean time spent, in the drift by freshwater shrimps, Gammarus pulex; (ii) to develop a drift distance–water velocity model from the experimental study, and validate it with field data; (iii) to examine the relationship between drift rate, water velocity and benthic density with the latter expressed as a mean value for the whole stream and a mean value corrected for the distance travelled in the drift. 2. In field experiments at 10 water velocities (0.032–0.962 m s^?1), the significant relationship between the mean drift distance and mean water velocity was described both by a power function (power, 0.96) and a linear relationship. The mean drift time was fairly constant at 8.3 s (95% CL ± 0.4). A simple model estimated the drift distance and time spent in the drift by different percentages of the drifting invertebrates. This model predicted correctly the positive relationship between drift rate and water velocity for field data over a year. 3. The relationship between drift rate per hour and the independent variables, water velocity and benthic density, was well described by a multiple‐regression model. Adding temperature and date did not improve model fit. Variations in water velocity and benthic density explained 96% of the variation in nocturnal drift rate (65% to velocity, 31% to benthic density), but only 40% of the variation in diurnal drift rate (29% to velocity, 11% to benthic density). Correcting benthic density for the drift distances did not improve model fit. 4. The significance of this study is that it developed models to predict drift distances and time, values being similar to those obtained in another, larger stream. It also illustrated the importance of spatial scale in the interpretation of drift by showing that when drift distances were taken into account, the impact of drift on the population was higher (4–10% lost day^?1) than when drift distances were ignored (usually < 3% lost day^?1), especially at a local level.     

Computer simulation of flagellar movement: VII. Conventional but functionally different cross-bridge models for inner and outer arm dyneins can explain the effects of outer arm dynein removal

Outer arm dynein removal from flagella by genetic or chemical methods causes decreased frequency and power, but little change in bending pattern. These results suggest that outer arm dynein operates within bends to increase the speed of bend propagation, but does not produce forces that alter the bending pattern established by inner arm dyneins. A flagellar model incorporating different cross-bridge models for inner and outer arm dyneins has been examined. The inner arm dynein model has a hyperbolic force-velocity curve, with a maximum average force at 0 sliding velocity of about 14 pN for each 96 nm group of inner arm dyneins. The outer arm dynein model has a very different force-velocity curve, with a maximum force at about 10-15% of V(max). The outer arm dynein model is adjusted so that the unloaded sliding velocity for a realistic mixture of inner and outer arm dyneins is twice the unloaded sliding velocity for the inner arm dynein model alone. With these cross-bridge models, a flagellar model can be obtained that reduces its sliding velocity and frequency by approximately 50% when outer arm dyneins are removed, with little change in bending pattern. The addition of outer arm dyneins, therefore, gives an approximately 4-fold increase in power output against viscous resistances, and outer arm dyneins may generate 90% or more of the power output. Cell Motil.     

Pathway of ultrasound waves in the equine third metacarpal bone

The velocity of ultrasound waves through bone has been used widely as a non-invasive method for assessing bone quality. Accurate measurement of velocity depends on accurate assessment of the distance travelled by the sound wave. It has been argued that the sonic pathway is deflected around the marrow cavity and so does not follow a straight line through the bone; therefore, correction factors have been developed to account for the extra distance travelled. This hypothesis was examined in vitro using sections from the equine third metacarpal bone. Two 1 MHz transducers used with the transmitting transducer energized by a 600 V electrical spike generator produced a 0.1 μs pulse width and the received signal was recorded on a delayed time-base oscilloscope, from which the velocity was calculated. Two distinct peaks were apparent in the received signal, corresponding to a direct cortical transmission wave and a direct medullary transmission wave. This observation was confirmed quantitatively using models of the third metacarpal made from homogeneous materials that allow accurate determination of the transit times of each component of the signal. Perspex was used to mimic cortical bone, with water as the mimic for the contents of the medullary canal; these materials were chosen because they have transmission velocities similar to the materials they were mimicing. The results confirmed that the pathway went straight through the bone with a time lag in the medullary wave due to the slower transmission velocity of the marrow. To ensure that the cortical wave is always received, transducers larger than the medullary width should always be used.     

Constipating and spasmolytic effects of Khat (Catha edulis Forsk) in experimental animals.

The constipating and spasmolytic effects of Catha edulis Forsk (Khat) were investigated in whole mice and on isolated guinea pig ileum. D-amphetamine was employed in both experiments for comparison. The total distance travelled (expressed in percentage) by charcoal suspension in the gastrointestinal tract of mice was determined before and after khat administration. The procedure was repeated with amphetamine and normal saline. The results were compared. Amplitudes of contraction were recorded with standard spasmogens, histamine and carbachol, in the presence and absence of khat extract of different concentrations. The same was done with amphetamine. Khat extract was observed to reduce the total distance travelled by charcoal suspension, comparable to D-amphetamine. The spasmogenic effects of both histamine and carbachol were observed to be antagonized by the khat extract in a concentration-dependent manner. The antispasmodic effect of khat extract was observed to be similar to that of D-amphetamine.     

Regulation of outer-arm-dynein activity by phosphorylation and control of ciliary beat frequency

Summary Ciliary beating is empowered by a mechanochemical enzyme, dynein, which appears as two rows of projections on doublet microtubules. While inner-arm dyneins modulate beat form, outer-arm dynein empowers ciliary beat and sets beat frequency. Beat frequency is controlled via phosphorylation of outer-arm dynein. UsingParamecium tetraurelia as model system, we have previously identified a regulatory light chain of outer-arm dynein (22S dynein), M_r29 (p29), whose phosphorylation is cAMP-dependent. The phosphorylation state of the p29 in 22 S dynein determines in vitro microtubule translocation velocity. Although in vitro phosphorylation of p29 takes place in a short time, the percent change ist significantly less than the percent change in dynein activation, or in ciliary beat frequency. A potential mechanism that explains how a few activated dyneins can change ciliary beating is discussed.     

Regulation of flagellar dynein by the axonemal central apparatus

Numerous studies indicate that the central apparatus, radial spokes, and dynein regulatory complex form a signaling pathway that regulates dynein activity in eukaryotic flagella. This regulation involves the action of several kinases and phosphatases anchored to the axoneme. To further investigate the role of the central apparatus in this signaling pathway, we have taken advantage of a microtubule-sliding assay to assess dynein activity in central apparatus defective mutants of Chlamydomonas. Axonemes isolated from both pf18 and pf15 (lacking the entire central apparatus) and from pf16 (lacking the C1 central microtubule) have reduced microtubule-sliding velocity compared with wild-type axonemes. Based on functional analyses of axonemes isolated from radial spokeless mutants, we hypothesized that inhibitors of casein kinase 1 (CK1) and cAMP dependent protein kinase (PKA) would rescue dynein activity and increase microtubule-sliding velocity in central pairless mutants. Treatment of axonemes isolated from both pf18 and pf16 with DRB, a CK1 inhibitor, but not with PKI, a PKA inhibitor, restored dynein activity to wild-type levels. The DRB-induced increase in dynein-driven microtubule sliding was inhibited if axonemes were first incubated with the phosphatase inhibitor, microcystin. Inhibiting CK1 in pf15 axonemes, which lack the central pair as well as PP2A [Yang et al., 2000: J. Cell Sci. 113:91-102], did not increase microtubule-sliding velocity. These data are consistent with a model in which the central apparatus, and specifically the C1 microtubule, regulate dynein through interactions with the radial spokes that ultimately alter the activity of CK1 and PP2A. These data are also consistent with localization of axonemal CK1 and PP2A near the dynein arms.     

Microtubule translocation properties of intact and proteolytically digested dyneins from Tetrahymena cilia

Tetrahymena cilia contain a three-headed 22S (outer arm) dynein and a single-headed 14S dynein. In this study, we have employed an in vitro assay of microtubule translocation along dynein-coated glass surfaces to characterize the motile properties of 14S dynein, 22S dynein, and proteolytic fragments of 22S dynein. Microtubule translocation produced by intact 22S dynein and 14S dynein differ in a number of respects including (a) the maximal velocities of movement; (b) the ability of 22S dynein but not 14S dynein to utilize ATP gamma S to induce movement; (c) the optimal pH and ionic conditions for movement; and (d) the effects of Triton X-100 on the velocity of movement. These results indicate that 22S and 14S dyneins have distinct microtubule translocating properties and suggest that these dyneins may have specialized roles in ciliary beating. We have also explored the function of the multiple ATPase heads of 22S dynein by preparing one- and two-headed proteolytic fragments of this three-headed molecule and examining their motile activity in vitro. Unlike the single-headed 14S dynein, the single-headed fragment of 22S dynein did not induce movement, even though it was capable of binding to microtubules. The two-headed fragment, on the other hand, translocated microtubules at velocities similar to those measured for intact 22S dynein (10 microns/sec). This finding indicates that the intact three-headed structure of 22S dynein is not essential for generating microtubule movement, which raises the possibility that multiple heads may serve some regulatory function or may be required for maximal force production in the beating cilium.     

A model for the coordinated stepping of cytoplasmic dynein

Cytoplasmic dynein play an important role in transporting various intracellular cargos by coupling their ATP hydrolysis cycle with their conformational changes. Recent experimental results showed that the cytoplasmic dynein had a highly variable stepping pattern including “hand-over-hand”, “inchworm” and “nonalternating-inchworm”. Here, we developed a model to describe the coordinated stepping patterns of cytoplasmic dynein, based on its working cycle, construction and the interaction between its leading head and tailing head. The kinetic model showed how change in the distance between the two heads influences the rate of cytoplasmic dynein under different stepping patterns. Numerical simulations of the distribution of step size and striding rate are in good quantitative agreement with experimental observations. Hence, our coordinated stepping model for cytoplasmic dynein successfully explained its diverse stepping patterns as a molecular motor. The cooperative mechanism carried out by the two heads of cytoplasmic dynein shed light on the strategies adopted by the cytoplasmic dynein in executing various functions.     

Travel distance and mass gain in wintering blackbirds

Birds that range over a large area will have a greater mass-dependent risk of predation than more sedentary birds. Birds that travel more may then reduce winter mass gain to compensate for the increased predation risk that greater travelling entails. I tested whether European blackbirds, Turdus merula, that travelled more in winter had a lower mass than more sedentary birds, independently of any confounding effects of food supply on both ranging behaviour and mass gain. I measured change in winter mass and amount of food eaten in conjunction with the distance that blackbirds travelled to a randomly sited mobile feeder. Blackbirds that travelled shorter distances (per trip and in total) and less often to the feeder had the highest mass midwinter relative to their spring mass. Blackbirds with a higher mean mass midwinter also travelled, on average, shorter distances to the feeder. The distance an individual blackbird travelled to the feeder at any one time was probably independent of the state of its daily energy reserves (how much of its daily total mass gain it had achieved at that point). The relationship between distance travelled and mass was probably independent of food supply because distances actually increased at the end of the winter and the amount of food eaten per individual changed little. More mobile blackbirds were therefore likely to have compensated for any increases in predation risk associated with their greater ranges by decreasing winter mass gain, but will have had an increased risk of starvation because of their lower mass. Copyright 1999 The Association for the Study of Animal Behaviour.     

A stochastic locomotor control model for the nurse shark,ginglymostoma cirratum

Summary The locomotor behavior of the nurse shark (Ginglymostoma cirratum) is characterized by 17 variables (frequency and ratios of left, right, and total turns; their radians; straight paths (steps); distance travelled; and velocity) Within each of these variables there is an internal time dependency the structure of which was elaborated together with an improved statistical model predicting their behavior within 90% confidence limits. The model allows for the sensitive detection of subtle locomotor response to sensory stimulation as values of variables may exceed the established confidence limits within minutes after onset of the stimulus. The locomotor activity is well described by an autoregression time series model and can be predicted by only seven variables. Six of these form two independently operating clusters. The first one consists of: the number of right turns, the distance travelled, and the mean velocity; the second one of: the mean size of right turns, of left turns, and of all turns. The same clustering is obtained independently by a cluster analysis of cross-sections of the seven time series. It is apparent that, among a total of 17 locomotor variables, seven behave as individually independent agents, presumably controlled by seven separate and independent centers. The output of each center can only be predicted by its own behavior. In spite of the individual behavior of the seven variables, their internal structure is similar in important aspects which may result from control by a common command center. The shark locomotor model differs in important aspects from that previously constructed for the goldfish. The interdependence of the locomotor variables in both species may be related to the control mechanisms postulated by von Holst for the coordination of rhythmic fin movements in fishes. A locomotor control model for the nurse shark is proposed.     

The distances travelled by downstream-moving trout fry, Salmo trutta, in a Lake District stream

SUMMARY. 1. Field experiments were performed in the day and night at six modal water velocities (range 10–52cm s⁻¹), using: (i) newly- emerged fry without neutral buoyancy; (ii) older fry in poor condition (weight well below that expected for resident fry); (iii) older fry in good condition (weight similar to that of resident fry); (iv) dead fry.
2. An exponential model described the return rate of fry to the stream bottom; the mean distance travelled downstream varied considerably between the four fry categories, but always increased linearly with increasing water velocity.
3. Results were similar for dead fry and newly-emerged fry released at night; 50% of the fry returned to the bottom in 10–11 s and nearly all returned in c . 70s, the maximum distance travelled ranging from c . 7 m at 10 cm s⁻¹ to c . 37m at 52cm s⁻¹, Newly-emerged fry released in the day returned slightly faster (54s for 99% return to bottom).
4. Older fry in poor condition returned to the bottom slightly faster in the day than at night, but took about 2 min and travelled about twice the distance covered by dead fry. Older fry in good condition returned to the bottom at the fastest rate (3–6s for 50% and c . 30s for the rest), and travelled only about half (at night) or a third (in day) of the distance covered by dead fry.
5. The implications of this investigation are discussed and it is concluded that, apart from water velocity, the age and condition of the fry were the two most important factors affecting their downstream movement.     

A Computational Model of Dynein Activation Patterns that Can Explain Nodal Cilia Rotation

Normal left-right patterning in vertebrates depends on the rotational movement of nodal cilia. In order to produce this ciliary motion, the activity of axonemal dyneins must be tightly regulated in a temporal and spatial manner; the specific activation pattern of the dynein motors in the nodal cilia has not been reported. Contemporary imaging techniques cannot directly assess dynein activity in a living cilium. In this study, we establish a three-dimensional model to mimic the ciliary ultrastructure and assume that the activation of dynein proteins is related to the interdoublet distance. By employing finite-element analysis and grid deformation techniques, we simulate the mechanical function of dyneins by pairs of point loads, investigate the time-variant interdoublet distance, and simulate the dynein-triggered ciliary motion. The computational results indicate that, to produce the rotational movement of nodal cilia, the dynein activity is transferred clockwise (looking from the tip) between the nine doublet microtubules, and along each microtubule, the dynein activation should occur faster at the basal region and slower when it is close to the ciliary tip. Moreover, the time cost by all the dyneins along one microtubule to be activated can be used to deduce the dynein activation pattern; it implies that, as an alternative method, measuring this time can indirectly reveal the dynein activity. The proposed protein-structure model can simulate the ciliary motion triggered by various dynein activation patterns explicitly and may contribute to furthering the studies on axonemal dynein activity.     

Quantifying the effects of habitat structure on prey detectability and accessibility to farmland birds

For species that rely on visual cues to detect prey items, increasing the structural complexity of a patch can greatly influence forager behaviour through consequent reductions in prey detectability and accessibility. These effects are likely to manifest themselves in terms of foraging site selection and there is plentiful evidence for preferential site selection for a suite of taxa. However, the underlying effects of habitat structure on foraging behaviour, which are likely to drive these observed site selections, are much less well understood. We present the results of two studies designed to quantify the effects of vegetation structure on prey detectability and accessibility to avian invertebrate feeders and granivores on farmland. There was a significant negative relationship between potential prey detectability and both distance and vegetation height in cereal crops and stubbles for Northern Lapwings Vanellus vanellus . The interscan distance travelled by Lapwings differed significantly between habitats, with longer distances travelled in cereal crops and harrowed compared with ploughed soil and grasses. The peck rate, head-up rate and mean search period of foraging Chaffinches Fringilla coelebs were not affected by increasing vegetation structure but forager mobility was significantly reduced. We hope that by quantifying the effects of vegetation structure on prey detectability and accessibility we can highlight the importance of considering these factors, as well as prey abundance, when developing management strategies for farmland birds.     

Organelle transport along microtubules in Xenopus melanophores: evidence for cooperation between multiple motors

Xenopus melanophores have pigment organelles or melanosomes which, in response to hormones, disperse in the cytoplasm or aggregate in the perinuclear region. Melanosomes are transported by microtubule motors, kinesin-2 and cytoplasmic dynein, and an actin motor, myosin-V. We explored the regulation of melanosome transport along microtubules in vivo by using a new fast-tracking routine, which determines the melanosome position every 10 ms with 2-nm precision. The velocity distribution of melanosomes transported by cytoplasmic dynein or kinesin-2 under conditions of aggregation and dispersion presented several peaks and could not be fit with a single Gaussian function. We postulated that the melanosome velocity depends linearly on the number of active motors. According to this model, one to three dynein molecules transport each melanosome in the minus-end direction. The transport in the plus-end direction is mainly driven by one to two copies of kinesin-2. The number of dyneins transporting a melanosome increases during aggregation, whereas the number of active kinesin-2 stays the same during aggregation and dispersion. Thus, the number of active dynein molecules regulates the net direction of melanosome transport. The model also shows that multiple motors of the same polarity cooperate during the melanosome transport, whereas motors of opposite polarity do not compete.     

Bending patterns of Chlamydomonas flagella: IV. Mutants with defects in inner and outer dynein arms indicate differences in dynein arm function

Mutants with outer dynein arm defects or deficiencies all show a major reduction in beat frequency to about half the normal value; some of these mutants show an additional decrease in sliding velocity associated with reduced shear amplitude and an additional reduction in beat frequency, as well as other more minor modifications of the normal forward mode bending pattern. New mutants (ida98, pf30), which appear to be deficient in a subset of inner dynein arms show a reduction in sliding velocity that is primarily associated with a reduction in shear amplitude, with only a small reduction in beat frequency. These differences in motility phenotype between inner and outer dynein arm mutants suggest that inner and outer dynein arms may have distinct functions. The relatively large decrease in sliding velocity associated with partial loss of inner arms is consistent with earlier observations on pf23, a nonmotile mutant lacking inner arms, suggesting that inner arms may have an essential function in motility. The ability to generate reverse mode bending patterns is retained in some inner or outer dynein arm mutants, but appears to be decreased in those mutants which show reduced shear amplitude for the forward mode bending pattern.     

The role of glandular and non-glandular trichomes in the negative interactions between strawberry cultivars and spider mite

In tropical countries, spider mite (Tetranychus urticae Koch) is a major pest of strawberries. This pest is mainly controlled by the application of pesticides. Use of pest-resistant cultivars is a healthy and environment-friendly alternative to pesticide use. This paper describes the role of glandular and non-glandular trichomes in the interaction between strawberry cultivars and spider mite. The methodology used in this study was based on two bioassays. First, the thumbtack bioassay of Weston and Snyder was used to differentiate strawberry cultivars in terms of the distance travelled by the mites. Second, different types of trichomes present on the abaxial surface of the strawberry leaves were identified and counted. The results of this study showed a significantly negative correlation between the distance travelled by the spider mites and the density of glandular trichomes on the strawberry plant. The cultivars Camino Real and Dover had the highest densities of glandular trichomes and the shortest distance travelled by the mites. In contrast, the cultivars Ventana and Toyonoka had the lowest density of glandular trichomes and the longest distance travelled by the spider mites. The high density of glandular trichomes might have been responsible for the spider mite non-preference to the Camino Real and Dover strawberry cultivars, whereas the non-glandular trichomes minimally contributed to this non-preference. The results of this study affirm the role of glandular trichomes in negative interactions between strawberry and spider mites.     

First Insight into Exploration and Cognition in Wild Caught and Domesticated Sea Bass (Dicentrarchus labrax) in a Maze

European sea bass aquaculture is so recent that very little is known on the effects of the early steps of its domestication. Behavioural parameters are sensitive indicators of the domestication process since they are generally impacted as soon as the first generation. The present work compared wild-caught and domesticated sea bass juvenile swimming activity, exploration and ability to learn to discriminate between two 2-D objects associated to a simple spatial task that enabled the tested individual to visually interact with an unfamiliar congener (the reward) located behind a transparent wall at the end of one of the two arms of a maze. Ten fish from each origin were individually tested 3 times in a row during 3 days (9 trials in total). Fish were placed in a start box closed by a transparent wall located in front of two 2-D objects. Fish were filmed during 10 min after the removal of the start box wall. Different swimming variables including angular velocity, total distance travelled and velocity mean, were analyzed from videos as well as the time spent in each of 6 virtual zones including the reward zone near the congener (Cong) and the zone opposite to the reward zone (OpCong). Two learning criteria were chosen: the number of successful turns and time to reach Cong. Behavioural differences were found between domesticated and wild fish. Angular velocity was higher in wild fish while the distance travelled and the velocity mean were higher in domesticated ones. Wild and domesticated fish spent most of the time in Cong and in OpCong. No differences were seen in learning ability between wild and domesticated fish. However, our findings for learning require confirmation by further studies with larger numbers of learning sessions and experiments designed to minimise stress. This study therefore demonstrated an impact of domestication on swimming behaviour but not on spatial learning.