Contractile equilibration of single cells to step changes in extracellular stiffness

Extracellular stiffness has been shown to alter long timescale cell behaviors such as growth and differentiation, but the cellular response to changes in stiffness on short timescales is poorly understood. By studying the contractile response of cells to dynamic stiffness conditions using an atomic force microscope, we observe a seconds-timescale response to a step change in extracellular stiffness. Specifically, we observe acceleration in contraction velocity (μm/min) and force rate (nN/min) upon a step decrease in stiffness and deceleration upon a step increase in stiffness. Interestingly, this seconds-timescale response to a change in extracellular stiffness is not altered by inhibiting focal adhesion signaling or stretch-activated ion channels and is independent of cell height and contraction force. Rather, the response timescale is altered only by disrupting cytoskeletal mechanics and is well described by a simple mechanical model of a constant velocity actuator pulling against an internal cellular viscoelastic network. Consistent with the predictions of this model, we find that an osmotically expanding hydrogel responds to step changes in extracellular stiffness in a similar manner to cells. We therefore propose that an initial event in stiffness sensing is establishment of a mechanical equilibrium that balances contraction of the viscoelastic cytoskeleton with deformation of the extracellular matrix.     

The cytoplasmic tyrosine kinase Arg regulates gastrulation via control of actin organization

Coordinated cell movements are crucial for vertebrate gastrulation and are controlled by multiple signals. Although many factors are shown to mediate non-canonical Wnt pathways to regulate cell polarity and intercalation during gastrulation, signaling molecules acting in other pathways are less investigated and the connections between various signals and cytoskeleton are not well understood. In this study, we show that the cytoplasmic tyrosine kinase Arg modulates gastrulation movements through control of actin remodeling. Arg is expressed in the dorsal mesoderm at the onset of gastrulation, and both gain- and loss-of-function of Arg disrupted axial development in Xenopus embryos. Arg controlled migration of anterior mesendoderm, influenced cell decision on individual versus collective migration, and modulated spreading and protrusive activities of anterior mesendodermal cells. Arg also regulated convergent extension of the trunk mesoderm by influencing cell intercalation behaviors. Arg modulated actin organization to control dynamic F-actin distribution at the cell-cell contact or in membrane protrusions. The functions of Arg required an intact tyrosine kinase domain but not the actin-binding motifs in its carboxyl terminus. Arg acted downstream of receptor tyrosine kinases to regulate phosphorylation of endogenous CrkII and paxillin, adaptor proteins involved in activation of Rho family GTPases and actin reorganization. Our data demonstrate that Arg is a crucial cytoplasmic signaling molecule that controls dynamic actin remodeling and mesodermal cell behaviors during Xenopus gastrulation.     

Molecular remodeling mechanisms of the neural somatodendritic compartment

Neuronal cells use the process of vesicle trafficking to manipulate the populations of neurotransmitter receptors and other membrane proteins. Long term potentiation (LTP) is a long-lived increase in synaptic strength between neurons and increases postsynaptic dendritic spine size and the concentration of the α-amino-3-hydroxy-5-methyl-4-isoxazole propionate-type glutamate receptor (AMPAR) located in the postsynaptic density. AMPAR is removed from the cell surface via clathrin-mediated endocytosis. While the adaptor protein 2 (AP2) complex of endocytosis seems to have the components needed to allow temporal and spatial regulations of internalization, many accessory proteins are involved, such as epidermal growth factor receptor phosphorylation substrate 15 (Eps15). A sequence of repeats in the Eps15 protein is known as the Eps15 homology (EH) domain. It has affinity for asparagine-proline-phenylalanine (NPF) sequences that are contained within vesicle trafficking proteins such as epsin, Rab11 family interacting protein 2 (Rab11-FIP2), and Numb. After endocytosis, a pool of AMPAR is stored in the endosomal recycling compartment that can be transported to the dendritic spine surface upon stimulation during LTP for lateral diffusion into the postsynaptic density. Rab11 and the Eps15 homologue EHD1 are involved in receptor recycling. EHD family members are also involved in transcytosis of the neuronal cell adhesion molecule neuron-glia cell adhesion molecule (NgCAM) from the somatodendritic compartment to the axon. Neurons have a unique morphology comprising many projections of membrane that is constructed in part by the effects of the Eps15 homologue, intersectin. Morphogenesis in the somatodendritic compartment is becoming better understood, but there is still much exciting territory to explore, especially regarding the roles of various EH domain-NPF interactions in endocytic and recycling processes.     

The Dopamine D1-D2 Receptor Heteromer Localizes in Dynorphin/Enkephalin Neurons: INCREASED HIGH AFFINITY STATE FOLLOWING AMPHETAMINE AND IN SCHIZOPHRENIA*

The distribution and function of neurons coexpressing the dopamine D1 and D2 receptors in the basal ganglia and mesolimbic system are unknown. We found a subset of medium spiny neurons coexpressing D1 and D2 receptors in varying densities throughout the basal ganglia, with the highest incidence in nucleus accumbens and globus pallidus and the lowest incidence in caudate putamen. These receptors formed D1-D2 receptor heteromers that were localized to cell bodies and presynaptic terminals. In rats, selective activation of D1-D2 heteromers increased grooming behavior and attenuated AMPA receptor GluR1 phosphorylation by calcium/calmodulin kinase IIα in nucleus accumbens, implying a role in reward pathways. D1-D2 heteromer sensitivity and functional activity was up-regulated in rat striatum by chronic amphetamine treatment and in globus pallidus from schizophrenia patients, indicating that the dopamine D1-D2 heteromer may contribute to psychopathologies of drug abuse, schizophrenia, or other disorders involving elevated dopamine transmission.     

Bacterial community responses to increasing ammonia concentrations in model recirculating vertical flow saline biofilters

This study represents the first analysis of ammonia removal and bacterial communities in gravel biofilters treating saline wastewater and is of relevance to the growing inland marine aquaculture industry. As part of a study to gain greater understanding of the microbial processes occurring in a newly constructed limestone gravel wetland at a commercial marine fish farm, this study was designed to establish the ammonia removal capacity of model biofilters treating saline aquaculture wastewater and to investigate changes to total bacterial communities and ammonia-oxidizing bacterial communities as the biofilters are exposed to increasing ammonia concentrations. Three replicate laboratory-scale gravel biofilters were constructed and the limits of nitrification capacity were tested by dosing with aquaculture wastewater supplemented with increasing amounts of ammonium chloride. The experiment was run over a 12-week period with the water temperature between 24.5 and 28 °C and salinity between 28 and 38 ppt. Greater than 97% ammonia removal in each weekly treatment period was observed with ammonia concentrations of up to 600 ppm. At higher concentrations of ammonia, a lower percentage of ammonia was removed, and on occasion nitrite accumulation was observed. A drop in the number of operational taxonomic units (OTUs) detected in the bacterial community as measured by 16s rRNA T-RFLP was observed concurrent with the decrease in percentage ammonia removal. T-RFLP of the amoA gene showed the experimental biofilters to be dominated by three different OTUs of ammonia-oxidizing bacteria. A synchronous successional pattern among these three ammonia oxidizers was observed. The three OTUs were identified as belonging to three different nitrosomonad clusters. This study demonstrates that the vertical flow gravel biofilters have the ability to treat saline aquaculture wastewater that has a high ammonia concentration and that the microbial community within saline biofilters has the capacity to adapt to changing ammonia levels while maintaining nitrification activity.     

Estimating overdispersion in sparse multinomial data

Multinomial data arise in many areas of the life sciences, such as mark-recapture studies and phylogenetics, and will often by overdispersed, with the variance being higher than predicted by a multinomial model. The quasi-likelihood approach to modeling this overdispersion involves the assumption that the variance is proportional to that specified by the multinomial model. As this approach does not require specification of the full distribution of the response variable, it can be more robust than fitting a Dirichlet-multinomial model or adding a random effect to the linear predictor. Estimation of the amount of overdispersion is often based on Pearson's statistic X² or the deviance D. For many types of study, such as mark-recapture, the data will be sparse. The estimator based on X² can then be highly variable, and that based on D can have a large negative bias. We derive a new estimator, which has a smaller asymptotic variance than that based on X², the difference being most marked for sparse data. We illustrate the numerical difference between the three estimators using a mark-recapture study of swifts and compare their performance via a simulation study. The new estimator has the lowest root mean squared error across a range of scenarios, especially when the data are very sparse.     

Temporal population variability in local forest communities has mixed effects on tree species richness across a latitudinal gradient

Among the local processes that determine species diversity in ecological communities, fluctuation‐dependent mechanisms that are mediated by temporal variability in the abundances of species populations have received significant attention. Higher temporal variability in the abundances of species populations can increase the strength of temporal niche partitioning but can also increase the risk of species extinctions, such that the net effect on species coexistence is not clear. We quantified this temporal population variability for tree species in 21 large forest plots and found much greater variability for higher latitude plots with fewer tree species. A fitted mechanistic model showed that among the forest plots, the net effect of temporal population variability on tree species coexistence was usually negative, but sometimes positive or negligible. Therefore, our results suggest that temporal variability in the abundances of species populations has no clear negative or positive contribution to the latitudinal gradient in tree species richness.     

Developing a system for in vivo imaging of maize roots containing iodinated contrast media in soil using synchrotron XCT and XRF

Plant and Soil - We sought to develop a novel experimental system which enabled application of iodinated contrast media to in vivo plant roots intact in soil and was compatible with time-resolved...     

Conspecific and Heterospecific Cues Override Resource Quality to Influence Offspring Production

Animals live in an uncertain world. To reduce uncertainty, animals use cues that can encode diverse information regarding habitat quality, including both non-social and social cues. While it is increasingly appreciated that the sources of potential information are vast, our understanding of how individuals integrate different types of cues to guide decision-making remains limited. We experimentally manipulated both resource quality (presence/absence of cactus fruit) and social cues (conspecific juveniles, heterospecific juveniles, no juveniles) for a cactus-feeding insect, Narnia femorata (Hemiptera: Coreidae), to ask how individuals responded to resource quality in the presence or absence of social cues. Cactus with fruit is a high-quality environment for juvenile development, and indeed we found that females laid 56% more eggs when cactus fruit was present versus when it was absent. However, when conspecific or heterospecific juveniles were present, the effects of resource quality on egg numbers vanished. Overall, N . femorata laid approximately twice as many eggs in the presence of heterospecifics than alone or in the presence of conspecifics. Our results suggest that the presence of both conspecific and heterospecific social cues can disrupt responses of individuals to environmental gradients in resource quality.