Model for Coordination of Microtubule and Actin Dynamics in Growth Cone Turning

In the developing nervous system, axons are guided to their synaptic targets by motile structures at the axon tip called growth cones, which reorganize their cytoskeleton in order to steer in response to chemotactic cues. Growth cone motility is mediated by an actin-adhesion “clutch” mechanism, in which mechanical attachment to a substrate, coupled with polarized actin growth, produces leading-edge protrusion. Several studies suggest that dynamic microtubules (MTs) in the growth cone periphery play an essential role in growth cone steering. It is not yet well-understood how the MT cytoskeleton and the dynamic actin-adhesion clutch system are coordinated to promote growth cone navigation. I introduce an experimentally motivated stochastic model of the dynamic reorganization of the growth cone cytoskeleton in response to external guidance cues. According to this model, asymmetric decoupling of MTs from actin retrograde flow leads to a local influx of MTs to the growth cone leading edge, and the leading-edge MT accumulation is amplified by positive feedback between MTs and the actin-adhesion clutch system. Local accumulation of MTs at the leading edge is hypothesized to increase actin adhesion to the substrate, which attenuates actin retrograde flow and promotes leading-edge protrusion. Growth cone alignment with the chemotactic gradient is predicted to be most effective for intermediate levels of sensitivity of the adhesion strength to the presence of leading-edge MTs. Quantitative predictions of the MT distribution and the local rate of retrograde actin flow will allow the hypothetical positive feedback mechanism to be experimentally tested

Realization of a feedback controlled flashing ratchet

A flashing ratchet transports diffusive particles using a time-dependent, asymmetric potential. Particle speed is predicted to increase when a feedback algorithm based on particle positions is used. We have experimentally realized such a feedback ratchet using an optical line trap, and observed that use of feedback increases velocity by up to an order of magnitude. We compare two different feedback algorithms for small particle numbers, and find good agreement with simulations. We also find that existing algorithms can be improved to be more tolerant to feedback delay times

Mechanical coupling in flashing ratchets

We consider the transport of rigid objects with internal structure in a flashing ratchet potential by investigating the overdamped behavior of a rod-like chain of evenly spaced point particles. In 1D, analytical arguments show that the velocity can reverse direction multiple times in response to changing the size of the chain or the temperature of the heat bath. The physical reason is that the effective potential experienced by the mechanically coupled objects can have a different symmetry than that of individual objects. All analytical predictions are confirmed by Brownian dynamics simulations. These results may provide a route to simple, coarse-grained models of molecular motor transport that incorporate an object's size and rotational degrees of freedom into the mechanism of transport.Comment: 9 pages, 10 figure

Polarity sorting of axonal microtubules: a computational study

We present a computational model to test a “polarity sorting” mechanism for microtubule (MT) organization in developing axons. We simulate the motor-based axonal transport of short MTs to test the hypothesis that immobilized cytoplasmic dynein motors transport short MTs with their plus ends leading, so “mal-oriented” MTs with minus-end-out are transported toward the cell body while “correctly” oriented MTs are transported in the anterograde direction away from the soma. We find that dynein-based transport of short MTs can explain the predominately plus-end-out polarity pattern of axonal MTs but that transient attachments of plus-end-directed motor proteins and nonmotile cross-linker proteins are needed to explain the frequent pauses and occasional reversals observed in live-cell imaging of MT transport. Static cross-linkers increase the likelihood of a stalled “tug-of-war” between retrograde and anterograde forces on the MT, providing an explanation for the frequent pauses of short MTs and the immobility of longer MTs. We predict that inhibition of the proposed static cross-linker will produce disordered transport of short MTs and increased mobility of longer MTs. We also predict that acute inhibition of cytoplasmic dynein will disrupt the polarity sorting of MTs by increasing the likelihood of “incorrect” sorting of MTs by plus-end-directed motors

Size matters: variations in seagrass seed size at local scales affects seed performance

Seed size can have an impact on angiosperm reproductive fitness. Ecological theory predicts plants that will produce larger seeds in stressful environments to increase the chances of seedling survival and numerous small seeds in favourable conditions to increase the number of recruits. We measured seed morphology of the seagrass Heterozostera nigricaulis from four populations under differing environmental conditions in South East Australia. Seed size and mass among sites showed consistent differences over four flowering seasons. Seeds from exposed, ephemeral meadows (Blairgowrie, Edwards Point) were 19%–53% heavier than those from larger, stable meadows at more sheltered sites (Swan Bay, Point Henry). Overall, heavier seeds from exposed sites performed better in germination experiments and persisted (remained viable) longer compared to small seeds from sheltered sites. Seeds from sheltered sites showed contrasting levels of seed performance. Small seeds from Swan Bay had the lowest germination but the proportion of viable seeds after 12 months were much higher (41%) than similar sized seeds from Point Henry (0%). There are clear life history benefits of large seeds that facilitate seed persistence and germination at exposed sites; however, the performance of smaller seeds varied between sites and may be a function of other site-specific advantages

Ethical dilemmas when using citizen science for early detection of invasive tree pests and diseases

The early detection of tree health pests and disease is an important component of biosecurity to protect the aesthetic, recreational and economic importance of trees, woodlands and forestry. Citizen science is valuable in supporting the early detection of tree pests and diseases. Different stakeholders (government, business, society and individual) will vary in their opinion of the balance between costs and benefits of early detection and consequent management, partly because many costs are local whereas benefits are felt at larger scales. This can create clashes in motivations of those involved in citizen science, thus leading to ethical dilemmas about what is good and responsible conduct for the use of citizen science. We draw on our experience of tree health citizen science to exemplify five dilemmas. These dilemmas arise because: the consequences of detection may locally be severe (e.g. the destruction of trees); knowledge of these impacts could lead to refusal to make citizen science reports; citizen science reports can be made freely, but can be costly to respond to; participants may expect solutions even if these are not possible; and early detection is (by definition) a rare event. Effective engagement and dialogue across stakeholders, including public stakeholders, is important to properly address these issues. This is vital to ensure the public’s long-term support for and trust in the use of citizen science for the early detection of tree pests and diseases

Cytoplasmic Dynein Transports Axonal Microtubules in a Polarity-Sorting Manner

Axonal microtubules are predominantly organized into a plus-end-out pattern. Here, we tested both experimentally and with computational modeling whether a motor-based polarity-sorting mechanism can explain this microtubule pattern. The posited mechanism centers on cytoplasmic dynein transporting plus-end-out and minus-end-out microtubules into and out of the axon, respectively. When cytoplasmic dynein was acutely inhibited, the bidirectional transport of microtubules in the axon was disrupted in both directions, after which minus-end-out microtubules accumulated in the axon over time. Computational modeling revealed that dynein-mediated transport of microtubules can establish and preserve a predominantly plus-end-out microtubule pattern as per the details of the experimental findings, but only if a kinesin motor and a static cross-linker protein are also at play. Consistent with the predictions of the model, partial depletion of TRIM46, a protein that cross-links axonal microtubules in a manner that influences their polarity orientation, leads to an increase in microtubule transport

Local and systemic in vivo responses to osseointegrative titanium nanotube surfaces

Orthopedic implants requiring osseointegration are often surface modified; however, implants may shed these coatings and generate wear debris leading to complications. Titanium nanotubes (TiNT), a new surface treatment, may promote osseointegration. In this study, in vitro (rat marrow-derived bone marrow cell attachment and morphology) and in vivo (rat model of intramedullary fixation) experiments characterized local and systemic responses of two TiNT surface morphologies, aligned and trabecular, via animal and remote organ weight, metal ion, hematologic, and nondecalcified histologic analyses. In vitro experiments showed total adherent cells on trabecular and aligned TiNT surfaces were greater than control at 30 min and 4 h, and cells were smaller in diameter and more eccentric. Control animals gained more weight, on average; however, no animals met the institutional trigger for weight loss. No hematologic parameters (complete blood count with differential) were significantly different for TiNT groups vs. control. Inductively coupled plasma mass spectrometry (ICP-MS) showed greater aluminum levels in the lungs of the trabecular TiNT group than in those of the controls. Histologic analysis demonstrated no inflammatory infiltrate, cytotoxic, or necrotic conditions in proximity of K-wires. There were significantly fewer eosinophils/basophils and neutrophils in the distal region of trabecular TiNT-implanted femora; and, in the midshaft of aligned TiNT-implanted femora, there were significantly fewer foreign body giant/multinucleated cells and neutrophils, indicating a decreased immune response in aligned TiNT-implanted femora compared to controls