Morphological Polarity Research Articles

A conserved function for Strabismus in establishing planar cell polarity in the ciliated ectoderm during cnidarian larval development

Functional and morphological planar cell polarity (PCP) oriented along the oral-aboral body axis is clearly evident in the ectoderm of torpedo-shaped planula larvae of hydrozoan cnidarians such as Clytia hemisphaerica. Ectodermal epithelial cells bear a single motile cilium the beating of which is coordinated between cells, causing directional swimming towards the blunt, aboral pole. We have characterised PCP during Clytia larval development and addressed its molecular basis. PCP is first detectable in ectodermal cells during gastrulation as coordinated basal body positioning, the ciliary root becoming consistently positioned on the oral side of the apical surface of the cell. At later stages, more pronounced structural polarity develops around the base of each cilium in relation to the cilia beating direction, including a characteristic asymmetric cortical actin organisation. Morpholino antisense oligonucleotide and mRNA injection studies showed that PCP development requires the Clytia orthologues of the core Fz-PCP pathway components Strabismus (CheStbm), Frizzled (CheFz1) and Dishevelled (CheDsh). Morpholinos targeting any of these components prevented ectodermal PCP, disrupted ciliogenesis and inhibited embryo elongation during gastrulation, which involves cell intercalation. We show that YFP-tagged CheStbm adopts a polarised intracellular distribution, localising preferentially to the aboral boundary of each cell, as has been demonstrated in Drosophila and some vertebrate PCP studies. Our findings in a cnidarian strongly suggest that the Fz-PCP pathway is a highly conserved and evolutionary ancient metazoan feature that is probably widely responsible for oriented swimming and/or feeding in relation to body axis in the many ciliated larval types found throughout the animal kingdom.

Nonmuscle myosin II regulation of lung epithelial morphology

The regulation of epithelial cell shape and orientation during lung branching morphogenesis is not clearly understood. Nonmuscle myosins regulate cell size, morphology, and planar cell polarity. Here, we test the hypothesis that nonmuscle myosin II (NM II) regulates lung epithelial morphology in a spatially restricted manner. Epithelial cell orientation at airway tips in fetal mouse lungs underwent a significant transformation at embryonic day (E) E17. Treatment of E15 lung explants with the NM II inhibitor blebbistatin increased airway branching, epithelial cell size, and the degree of anisotropy in epithelial cells lining the airway stalks. In cultured MLE-12 lung epithelial cells, blebbistatin increased cell velocity, but left the migratory response to FGF-10 unchanged. In the developing lung, NM II acts to constrain cell morphology and orientation, but may be suppressed at sites of branching and cell migration. The regulation of epithelial orientation may therefore undergo dynamic variations from E15 to E17.

Cadherin-related protein 24 induces morphological changes and partial cell polarization by facilitating direct cell-cell interactions

Cadherin-related protein 24 (CDHR24) is a potential tumor suppressor located apically as well as laterally in polarized cells. Here, the role of CDHR24 in contributing to cell morphology and polarity is examined. CDHR24 was predominantly localized at the nonattached part of nonpolarizing cells while another apically sorted protein, aminopeptidase N, was equally distributed over the plasma membrane. Furthermore, CDHR24 expression induced cell aggregation capacity, indicating direct cell-cell interaction. The transepithelial resistance, however, was elevated in polarized MDCK cells, but not in nonpolarizing CHO cells. Our data propose a model in which CDHR24 is directly involved in cell and tissue morphogenesis.

Lipid raft redistribution and morphological cell polarization are separable processes providing a basis for hematopoietic stem and progenitor cell migration

Freshly isolated human hematopoietic stem and progenitor cells (HSPCs) are small and round cells which upon cultivation adopt a polarized morphology and redistribute certain cell surface antigens. To functionally dissect this polarization process, we addressed impacts of protein synthesis, HSPC trafficking, cytoskeleton organization or lipid raft integrity on the establishment and maintenance of the cell polarity of human HSPCs. Effects on the morphology, sub-cellular distribution of lipid raft-associated molecular polarization markers (Flotillin-1, Flotillin-2, ICAM-3) and in vitro migration capabilities of treated cells were studied. We could distinguish two levels of cellular polarization, a molecular and a morphological level. Our data suggest that protein synthesis, lipid raft integrity and enzymatic activities of PI3K and aPKC are required to organize the molecular cell polarity. The morphological cell polarization process, however, also depends on actin polymerization and rho-GTPase activities. In summary, our data qualify HSPC polarization processes as new pharmaceutical target to interfere with migratory and with homing capabilities of HSPCs.

A three-component mechanism for fibroblast migration with a contractile cell body that couples a myosin II–independent propulsive anterior to a myosin II–dependent resistive tail

To understand the mechanism of cell migration, we cultured fibroblasts on micropatterned tracks to induce persistent migration with a highly elongated morphology and well-defined polarity, which allows microfluidic pharmacological manipulations of regional functions. The function of myosin II was probed by applying inhibitors either globally or locally. Of interest, although global inhibition of myosin II inhibited tail retraction and caused dramatic elongation of the posterior region, localized inhibition of the cell body inhibited nuclear translocation and caused elongation of the anterior region. In addition, local application of cytochalasin D at the tip inhibited frontal extension without inhibiting forward movement of the cell nucleus, whereas local treatment posterior to the nucleus caused reversal of nuclear movement. Imaging of cortical dynamics indicated that the region around the nucleus is a distinct compression zone where activities of anterior and posterior regions converge. These observations suggest a three-component model of cell migration in which a contractile middle section is responsible for the movement of a bulky cell body and the detachment/retraction of a resistive tail, thereby allowing these regions to undergo coordinated movement with a moving anterior region that carries little load.

Study of Alloys Addition to the Electrodeposited Nanocrystalline Cobalt

Cobalt and its alloy have been identified as potential candidates for replacing hexavalent Chromium plating in corrosion resistant coating in acidic environment. In this study, the effect of alloys addition towards elemental composition, crystallographic structure characterization, surface morphology, hardness and potentiodynamic polarization of the cobalt alloys coatings is reported. Addition of Nickel (Ni) and Iron (Fe) to the Cobalt (Co) coatings are deposited on stainless steel substrate by electrodeposition method. The deposition is performed at acidic environment of pH 3. The granule sizes of cobalt alloys prepared by electrodepositionmethod are in the range of 34.95 nm72.08 nm. The microhardness of CoNiFe is the highest (267.8 HV) compared to Co and CoFe. CoNiFeperforms the smallest corrosion rate with 1.322 mmpy. It is found thatthe addition of Ni and Fe into pure cobaltimproves the hardness and corrosion behavior.

N-cadherin specifies first asymmetry in developing neurons

The precise polarization and orientation of developing neurons is essential for the correct wiring of the brain. In pyramidal excitatory neurons, polarization begins with the sprouting of opposite neurites, which later define directed migration and axo-dendritic domains. We here show that endogenous N-cadherin concentrates at one pole of the newborn neuron, from where the first neurite subsequently emerges. Ectopic N-cadherin is sufficient to favour the place of appearance of the first neurite. The Golgi and centrosome move towards this newly formed morphological pole in a second step, which is regulated by PI3K and the actin/microtubule cytoskeleton. Moreover, loss of function experiments in vivo showed that developing neurons with a non-functional N-cadherin misorient their cell axis. These results show that polarization of N-cadherin in the immediate post-mitotic stage is an early and crucial mechanism in neuronal polarity.

Excitatory Cortical Neurons with Multipolar Shape Establish Neuronal Polarity by Forming a Tangentially Oriented Axon in the Intermediate Zone

The formation of axon-dendrite polarity is crucial for neuron to make the proper information flow within the brain. Although the processes of neuronal polarity formation have been extensively studied using neurons in dissociated culture, the corresponding developmental processes in vivo are still unclear. Here, we illuminate the initial steps of morphological polarization of excitatory cortical neurons in situ, by sparsely labeling their neuroepithelial progenitors using in utero electroporation and then examining their neuronal progeny in brain sections and in slice cultures. Morphological analysis showed that an axon-like long tangential process formed in progeny cells in the intermediate zone (IZ). Time-lapse imaging analysis using slice culture revealed that progeny cells with multipolar shape, after alternately extending and retracting their short processes for several hours, suddenly elongated a long process tangentially. These cells then transformed into a bipolar shape, extending a pia-directed leading process, and migrated radially leaving the tangential process behind, which gave rise to an "L-shaped" axon. Our findings suggest that neuronal polarity in these cells is established de novo from a nonpolarized stage in vivo and indicate that excitatory cortical neurons with multipolar shape in the IZ initiate axon outgrowth before radial migration into the cortical plate.

Differential response of Drosophila cell lines to extracellular adenosine

Adenosine (Ado) is a crucial metabolite that affects a wide range of physiological processes. Key proteins regulating Ado signaling, transport and metabolism are conserved among vertebrates and invertebrates. It is well known that Ado influences proliferation of several vertebrate and invertebrate cells. Here we show that Ado negatively influences viability, changes morphology and mitochondrial polarity of the Drosophila imaginal disc cell line (Cl.8+) via a mechanism exclusively dependent on cellular Ado uptake. High transport of Ado is followed by phosphorylation and ATP production as a part of Ado salvation, which at higher concentrations may interfere with cellular homeostasis. In contrast, hematopoietic cell line Mbn2, which grows well in high Ado concentration, preferentially uses adenosine deaminase as a part of the purine catabolic pathway. Our results show that different types of Drosophila cell lines use different pathways for Ado conversion and suggest that such differences may be an important part of complex mechanisms maintaining energy homeostasis in the body.

Endogenous Ion Dynamics in Cell Motility and Tissue Regeneration

Directional cell migration is an essential process, including regeneration of tissues, wound healing, and embryonic development. Cells achieve persistent directional migration by polarizing the spatiotemporal components involved in the morphological polarity. Ion transporter proteins situated at the cell membrane generates small electric fields that can induce directional cell motility. Besides them, externally applied direct current electric fields induce similar kind of responses as cell orientation and directional migration. However, the bioelectric mechanisms that lead to cellular directedness are poorly understood. Therefore, understanding the bioelectric signaling cues can serve as a powerful modality in controlling the cell behaviour, which can contribute additional insights for development and regeneration.

Abstract 9410: A Simple and Accurate Algorithm for Localizing Accessory Pathways in Patients with Wolff-Parkinson-White Syndrome

Introduction: Delta wave morphology reflects the ventricular attachment site of accessory pathway (AP). Electrocardiographic R/S ratio and QRS polarity also depend on the AP location. Many algorithms for localizing APs are based on the delta wave morphology, R/S ratio, and QRS polarity. However, they are slightly complicated and it is sometimes difficult to determine the delta wave morphology accurately. In this study, we propose a new and simple algorithm for accurate localization of APs using only the R/S ratio. Methods: This study consisted of 101 patients with a single anterogradely conducting AP on a 12-lead ECG. R/S ratios in leads V1, V2 and aVF were analyzed. A new algorithm was developed by correlating the 12-lead ECG findings with successful ablation sites in the 40 initial consecutive patients. This algorithm was subsequently tested in the 61 residual consecutive patients. AP locations were divided into 5 regions based on fluoroscopical anatomy. Results: The new algorithm and the validation results were shown (figure). Among 61 patients, all 37 patients with left free wall APs showed the R/S ratio in lead V1 ≥ 0.5 (sensitivity 100%, specificity 100%). Left anterior and lateral APs were associated with the R/S ratio in lead aVF ≥ 1 (sensitivity 97%, specificity 100%). If the R/S ratio in lead V1 was < 0.5 and the R/S ratio in lead V2 was ≥ 0.5, the AP location was midseptal or posteroseptal (sensitivity 100%, specificity 94%). If the R/S ratio in both leads V1 and V2 was < 0.5, it was right free wall or right anteroseptal (sensitivity 89%, specificity 100%). Right anterior, lateral, and anteroseptal APs were associated with the R/S ratio in lead aVF ≥ 1 (sensitivity 98%, specificity 93%). Overall sensitivity of this method was 93% and specificity was 99%. The positive and negative predictive values were 96% and 98%, respectively. Conclusions: This novel algorithm using only the R/S ratio in leads V1, V2 and aVF provides a simple and accurate way to identify the AP localization.

Effect of Magnesium Ion on the Zinc Electrodeposition from Acidic Sulfate Electrolyte

The effects of Mg2+ ion on the zinc electrodeposition were systematically investigated in sulfuric acid solution through the characterizations of current efficiency (CE), power consumption (PC), deposit morphology, cathodic polarization, and cyclic voltammetry. The results demonstrate that there is no significant influence on CE and PC in the Mg2+ concentration range of 1 to 10 g L−1, but with a drastic decrease of the CE and rapid increase of PC at Mg2+ ion concentration above 15 g L−1. Based on the morphology observation and polarization curves, the presence of Mg2+ ions could also induce the coarse surface on the electrodeposited zinc accompanying the enhancement of the cathodic polarization, which becomes more distinct at a high concentration above 15 g L−1. Furthermore, hydrogen evolution could be promoted with the existence of Mg2+ ions according to cyclic voltammograms.

Water maser polarization of the water fountains IRAS 15445−5449 and IRAS 18043−2116

We present the morphology and linear polarization of the 22-GHz H2O masers in the high-velocity outflow of two post-AGB sources, d46 (IRAS 15445-5449) and b292 (IRAS 18043-2116). The observations were performed using The Australia Telescope Compact Array. Different levels of saturated maser emission have been detected for both sources. We also present the mid-infrared image of d46 overlaid with the distribution of the maser features that we have observed in the red-shifted lobe of the bipolar structure. The relative position of the observed masers and a previous radio continuum observation suggests that the continnum is produced along the blue-shifted lobe of the jet. It is likely due to synchrontron radiation, implying the presence of a strong magnetic field in the jet. The fractional polarization levels measured for the maser features of d46 indicate that the polarization vectors are tracing the poloidal component of the magnetic field in the emitting region. For the H2O masers of b292 we have measured low levels of fractional linear polarization. The linear polarization in the H2O maser region of this source likely indicates a dominant toroidal or poloidal magnetic field component. Since circular polarization was not detected it is not possible to determine the magnetic field strength. However, we present a 3-sigma evaluation of the upper limit intensity of the magnetic field in the maser emitting regions of both observed sources.

Effect of oxygen pressure on the grain and domain structure of polycrystalline 0.85PbMg1/3Nb2/3O3–0.15PbTiO3 thin films studied by scanning probe microscopy

0.85PbMg1/3Nb2/3O3–0.15PbTiO3 ferroelectric–relaxor thin films have been deposited on La0.5Sr0.5CoO3/(1 1 1) Pt/TiO2/SiO2/Si by pulsed laser ablation at various oxygen partial pressures in the range 0.05 to 0.4 Torr. All the films have a rhombohedral perovskite structure. The grain morphology and orientation are drastically affected by the oxygen pressure, studied by x-ray diffraction and scanning electron microscopy. The domain structure investigations by dynamic contact electrostatic force microscopy have revealed that the distribution of polar nanoregions and their dynamics is influenced by the grain morphology, orientation and more importantly, oxygen vacancies. The correlation length extracted from autocorrelation function images has shown that the polarization disorder decreases with oxygen pressure up to 0.3 Torr. The presence of polarized domains and their electric field induced switching is discussed in terms of internal bias field and domain wall pinning. Film deposited at 0.4 Torr presents a curious case with unique triangular grain morphology and large polarization disorder.

A pharmacological cocktail for arresting actin dynamics in living cells

The actin cytoskeleton is regulated by factors that influence polymer assembly, disassembly, and network rearrangement. Drugs that inhibit these events have been used to test the role of actin dynamics in a wide range of cellular processes. Previous methods of arresting actin rearrangements take minutes to act and work well in some contexts, but can lead to significant actin reorganization in cells with rapid actin dynamics, such as neutrophils. In this paper, we report a pharmacological cocktail that not only arrests actin dynamics but also preserves the structure of the existing actin network in neutrophil-like HL-60 cells, human fibrosarcoma HT1080 cells, and mouse NIH 3T3 fibroblast cells. Our cocktail induces an arrest of actin dynamics that initiates within seconds and persists for longer than 10 min, during which time cells maintain their responsivity to external stimuli. With this cocktail, we demonstrate that actin dynamics, and not simply morphological polarity or actin accumulation at the leading edge, are required for the spatial persistence of Rac activation in HL-60 cells. Our drug combination preserves the structure of the existing cytoskeleton while blocking actin assembly, disassembly, and rearrangement, and should prove useful for investigating the role of actin dynamics in a wide range of cellular signaling contexts.

H2O maser polarization of the water fountains IRAS 15445–5449 and IRAS 18043–2116

AbstractWe present the morphology and linear polarization of the 22-GHz H2O masers in the high-velocity outflow of two post-AGB sources, d46 (IRAS 15445–5449) and b292 (IRAS 18043–2116). The observations were performed using The Australia Telescope Compact Array. Different levels of saturated maser emission have been detected for both sources. We also present the mid-infrared image of d46 overlaid with the distribution of the maser features that we have observed in the red-shifted lobe of the bipolar structure. The relative position of the observed masers and a previous radio continuum observation suggests that the continnum is produced along the blue-shifted lobe of the jet. It is likely due to synchrontron radiation, implying the presence of a strong magnetic field in the jet. The fractional polarization levels measured for the maser features of d46 indicate that the polarization vectors are tracing the poloidal component of the magnetic field in the emitting region. For the H2O masers of b292 we have measured low levels of fractional linear polarization. The linear polarization in the H2O maser region of this source likely indicates a dominant toroidal or poloidal magnetic field component.

Axon selection

The shape of a neuron supplies valuable clues as to its function. Neurons typically extend a single long, thin axon, which will transmit signals, and several shorter and thicker dendrites, which will receive signals. The understanding of the means by which neurons acquire a polarized morphology is a fundamental issue in developmental neurobiology. The current view suggests that axon selection involves a stochastic mechanism. However, new data suggest that a polarized cytoplasm not only determines the position of neurite emergence, but also sets the conditions for morphological polarization. In vertebrates, neurons migrate before establishing their final morphology. Recent work shows that the polarized cytoplasm also determines how neurons migrate. Thus, neuronal migration might influence the processes by which neurons form an axon.

Conformational Switching in Ezrin Regulates Morphological and Cytoskeletal Changes Required for B Cell Chemotaxis

B cell chemotaxis occurs in response to specific chemokine gradients and is critical for homeostasis and immune response. The molecular regulation of B cell membrane-actin interactions during migration is poorly understood. In this study, we report a role for ezrin, a member of the membrane-cytoskeleton cross-linking ezrin-radixin-moesin proteins, in the regulation of the earliest steps of B cell polarization and chemotaxis. We visualized chemokine-induced changes in murine B cell morphology using scanning electron microscopy and spatiotemporal dynamics of ezrin in B cells using epifluorescence and total internal reflection microscopy. Upon chemokine stimulation, ezrin is transiently dephosphorylated to assume an inactive conformation and localizes to the lamellipodia. B cells expressing a phosphomimetic conformationally active mutant of ezrin or those in which ezrin dephosphorylation was pharmacologically inhibited displayed impaired microvillar dynamics, morphological polarization, and chemotaxis. Our data suggest a 2-fold involvement of ezrin in B cell migration, whereby it first undergoes chemokine-induced dephosphorylation to facilitate membrane flexibility, followed by relocalization to the actin-rich lamellipodia for dynamic forward protrusion of the cells.

Conservation of vestibular afferent segregation within the otolith endorgans at the striola across vertebrate classes

Hair cells are the basic units of transduction in the vestibular apparatus. An intriguing feature of the hair cells in the utricle and saccule is the reversal of their morphological polarity at the striola: the kinocilia of all hair cells lie toward the striola in the utricle, and away from it in the saccule. In a previous study we showed that, in mice, the striola demarcates afferent fibers projecting to the brainstem and cerebellum. We used retrograde transport of lipophilic tracers to determine whether the striola in the chick also separates afferent domains projecting to the brainstem and cerebellum. Here, we found that in the utricle, afferents projecting to the cerebellum originate lateral to the striola, whereas afferents to the vestibular nuclei in the brainstem originates medial to it. In the saccule, afferents projecting to cerebellum originate from an area superior to the striola, whereas afferents projecting to the vestibular nuclei are inferior to it. In the lagena, the cerebellar afferents originate predominantly anterior to the striola, whereas the afferents to the vestibular nuclei originate predominantly posterior to it. As in mice, these findings indicate that, the striola demarcates not only the reversal of hair cell polarity, but also the segregation of afferents to the brainstem and cerebellum. This implies a conservation of primary macular afferent pathways in several classes of vertebrates.

In Chemotaxing Fibroblasts, Both High-Fidelity and Weakly Biased Cell Movements Track the Localization of PI3K Signaling

Cell movement biased by a chemical gradient, or chemotaxis, coordinates the recruitment of cells and collective migration of cell populations. During wound healing, chemotaxis of fibroblasts is stimulated by platelet-derived growth factor (PDGF) and certain other chemoattractants. Whereas the immediate PDGF gradient sensing response has been characterized previously at the level of phosphoinositide 3-kinase (PI3K) signaling, the sensitivity of the response at the level of cell migration bias has not yet been studied quantitatively. In this work, we used live-cell total internal reflection fluorescence microscopy to monitor PI3K signaling dynamics and cell movements for extended periods. We show that persistent and properly aligned (i.e., high-fidelity) fibroblast migration does indeed correlate with polarized PI3K signaling; accordingly, this behavior is seen only under conditions of high gradient steepness (>10% across a typical cell length of 50 μm) and a certain range of PDGF concentrations. Under suboptimal conditions, cells execute a random or biased random walk, but nonetheless move in a predictable fashion according to the changing pattern of PI3K signaling. Inhibition of PI3K during chemotaxis is accompanied by loss of both cell-substratum contact and morphological polarity, but after a recovery period, PI3K-inhibited fibroblasts often regain the ability to orient toward the PDGF gradient.