Rapid and Robust Analysis of Cellular and Molecular Polarization Induced by Chemokine Signaling

Cells respond to chemokine stimulation by losing their round shape in a process called polarization, and by altering the subcellular localization of many proteins. Classic imaging techniques have been used to study these phenomena. However, they required the manual acquisition of many cells followed by time consuming quantification of the morphology and the co-localization of the staining of tens of cells. Here, a rapid and powerful method is described to study these phenomena on samples consisting of several thousands of cells using an imaging flow cytometry technology that combines the advantages of a microscope with those of a cytometer. Using T lymphocytes stimulated with CCL19 and staining for MHC Class I molecules and filamentous actin, a gating strategy is presented to measure simultaneously the degree of shape alterations and the extent of co-localization of markers that are affected by CCL19 signaling. Moreover, this gating strategy allowed us to observe the segregation of filamentous actin (at the front) and phosphorylated Ezrin-Radixin-Moesin (phospho-ERM) proteins (at the rear) in polarized T cells after CXCL12 stimulation. This technique was also useful to observe the blocking effect on polarization of two different elements: inhibition of actin polymerization by a pharmacological inhibitor and expression of mutants of the Par6/atypical PKC signaling pathway. Thus, evidence is shown that this technique is useful to analyze both morphological alterations and protein redistributions.     

Using digital image analysis to quantify the architectural parameters of roots grown in thin rhizotrons

Abstract

Roots grown in small chambers or rhizotrons combined with image analysis could be effective for screening traits of root morphology and architecture prior to field experimentation. The objective of this study was to evaluate and compare digital images of root systems with conventional root scans (RSCN) for several sorghum varieties grown in rhizotrons. Plants were grown in duplicate in slim (1.5‐cm thick) rhizotrons angled 15° to the vertical, filled with sandy topsoil and subsoil at field capacity (FC) or 50% available water content (AWC) in a split‐plot design in a glasshouse (16/24°C). Root growth visible on the glass surface was recorded weekly, and roots were washed out 42 and 90 days after sowing for digital imaging of the root systems on pin boards (PBI). The roots were then sub‐sampled to quantify the baseline root parameters from digital RSCN using WinRhizo® software with a flat‐bed scanner. Rhizotron images revealed little of the root system (1–2%) but PBI “recovered” up to 70% of the total root length (TRL) measured by RSCN. A high a priori image contrast, increased image resolution (9.1–12 megapixels) and optimized contrast threshold between roots and background were the key parameters in quantifying the roots from digital images. PBI revealed significant differences in rooting patterns, especially distribution of root length density (RLD) in the profile, which is useful for selecting varieties with improved resource capture. However, the root diameters estimated in the PBI were significantly larger than those measured by RSCN due to lack of contrast, root clustering and overlay.     

Coding and decoding with dendrites

Since the discovery of complex, voltage dependent mechanisms in the dendrites of multiple neuron types, great effort has been devoted in search of a direct link between dendritic properties and specific neuronal functions. Over the last few years, new experimental techniques have allowed the visualization and probing of dendritic anatomy, plasticity and integrative schemes with unprecedented detail. This vast amount of information has caused a paradigm shift in the study of memory, one of the most important pursuits in Neuroscience, and calls for the development of novel theories and models that will unify the available data according to some basic principles. Traditional models of memory considered neural cells as the fundamental processing units in the brain. Recent studies however are proposing new theories in which memory is not only formed by modifying the synaptic connections between neurons, but also by modifications of intrinsic and anatomical dendritic properties as well as fine tuning of the wiring diagram. In this review paper we present previous studies along with recent findings from our group that support a key role of dendrites in information processing, including the encoding and decoding of new memories, both at the single cell and the network level.     

Ontogeny of head and caudal fin shape of an apex marine predator: The tiger shark (Galeocerdo cuvier)

How morphology changes with size can have profound effects on the life history and ecology of an animal. For apex predators that can impact higher level ecosystem processes, such changes may have consequences for other species. Tiger sharks (Galeocerdo cuvier) are an apex predator in tropical seas, and, as adults, are highly migratory. However, little is known about ontogenetic changes in their body form, especially in relation to two aspects of shape that influence locomotion (caudal fin) and feeding (head shape). We captured digital images of the heads and caudal fins of live tiger sharks from Southern Florida and the Bahamas ranging in body size (hence age), and quantified shape of each using elliptical Fourier analysis. This revealed changes in the shape of the head and caudal fin of tiger sharks across ontogeny. Smaller juvenile tiger sharks show an asymmetrical tail with the dorsal (upper) lobe being substantially larger than the ventral (lower) lobe, and transition to more symmetrical tail in larger adults, although the upper lobe remains relatively larger in adults. The heads of juvenile tiger sharks are more conical, which transition to relatively broader heads over ontogeny. We interpret these changes as a result of two ecological transitions. First, adult tiger sharks can undertake extensive migrations and a more symmetrical tail could be more efficient for swimming longer distances, although we did not test this possibility. Second, adult tiger sharks expand their diet to consume larger and more diverse prey with age (turtles, mammals, and elasmobranchs), which requires substantially greater bite area and force to process. In contrast, juvenile tiger sharks consume smaller prey, such as fishes, crustaceans, and invertebrates. Our data reveal significant morphological shifts in an apex predator, which could have effects for other species that tiger sharks consume and interact with. J. Morphol. 277:556–564, 2016. © 2016 Wiley Periodicals, Inc.     

Variation in human limb joint articular morphology

Objectives

Synovial joints in human limbs strike a balance between mobility, stability, and articular fit, yet little is known about how these conflicting demands pattern intraspecific variation in articular shape. In this study, we use geometric morphometrics to establish the apportionment and magnitude of morphological variance of the articular surfaces of the human shoulder, elbow, hip, and knee. We hypothesize that variances will be comparable between articulating surfaces within a joint and will be larger in joints with smaller ranges of motion, given their plurality of functional demands.

Materials and Methods

Three-dimensional landmarks were taken on the articular surfaces of the glenohumeral, humeroulnar, acetabulofemoral, and tibiofemoral joints from CT scans of 200 skeletons from the University of Tennessee Donated Skeletal Collection (84 females, 116 males). Root mean-squared distances between articulations calculated from Procrustes shape coordinates were used to determine variance distributions.

Results

We found no difference in variances for each articular surface between the sexes or between left and right articular surfaces. A high range of motion is associated with greater morphological variance; however, this pattern is largely driven by the concave articular surfaces of each joint, which consistently exhibit statistically greater variance than their convex counterparts.

Discussion

The striking pattern of differential variance between articulating morphologies points to potential disparities in development between them. Consistently higher variance in concave surfaces may relate to chondral modeling theory for the formation of joints. Establishing intraspecific morphological variance patterns is a first step in understanding coordinated evolution among articular features.     

Morphological variation in populations of the diatom Asterionella ralfsii W. Smith from Nova Scotia,Canada

Preliminary investigations of the diatom genus Asterionella ralfsii W. Smith from Kejimkujik National Park, Nova Scotia indicate that its morphology differs from other reported forms. Mean cell length increased between the spring and the fall. Bimodal distribution of length classes occurred in several lakes and was not related to measured environmental variables. The need for further work on this species in this and other geographical areas is considered.     

Higher investment in flight morphology does not trade off with fecundity estimates in a poleward range‐expanding damselfly

1. Evolutionary increases in dispersal‐related traits are frequently documented during range expansions. Investment in flight‐related traits is energetically costly and a trade‐off with fecundity may be expected during range expansion. 2. However, in contrast to wing‐dimorphic species, this trade‐off is not general in wing‐monomorphic species. In the absence of a dispersal‐‐fecundity trade‐off, an increased investment in clutch size at the expansion front is expected possibly at a cost of reduced offspring size. 3. The study evaluated investment in female flight morphology and fecundity‐related traits (clutch size, hatchling size) and potential trade‐offs among these traits in replicated populations of the poleward range‐expanding damselfly Coenagrion scitulum. 4. Females at the expansion front had a higher relative thorax length, indicating an increased investment in flight; this can be explained by spatial sorting of dispersal ability or in situ natural selection at the expansion front. Edge females produced larger hatchlings, however, this pattern was totally driven by the population‐specific thermal larval regimes and could not be attributed to the range expansion per se. By contrast, clutch sizes did not differ between core and edge populations. There was no signal of a dispersal–fecundity trade‐off either for a trade‐off between clutch size and hatchling size. 5. These results indicate that evolution of a higher dispersal ability at the expansion front of C. scitulum does not trade off with investment in fecundity, hence a dispersal–fecundity trade‐off is unlikely to slow down range expansion of this species.     

Structural and physiological responses of two invasive weeds, <Emphasis Type="Italic">Mikania micrantha</Emphasis> and <Emphasis Type="Italic">Chromolaena odorata</Emphasis>, to contrasting light and soil water conditions

To better understand the requirement of light and soil water conditions in the invasion sites of two invasive weeds, Mikania micrantha and Chromolaena odorata, we investigated their structural and physiological traits in response to nine combined treatments of light [full, medium and low irradiance (LI)] and soil water (full, medium and low field water content) conditions in three glasshouses. Under the same light conditions, most variables for both species did not vary significantly among different water treatments. Irrespective of water treatment, both species showed significant decreases in maximum light saturated photosynthetic rate (P _max), photosynthetic nitrogen-use efficiency, and relative growth rate under LI relative to full irradiance; specific leaf area, however, increased significantly from full to LI though leaf area decreased significantly, indicating that limited light availability under extreme shade was the critical factor restricting the growth of both species. Our results also indicated that M. micrantha performed best under a high light and full soil water combination, while C. odorata was more efficient in growth under a high light and medium soil water combination.