GAR22β regulates cell migration, sperm motility, and axoneme structure

© 2016 Gamper et al. Spatiotemporal cytoskeleton remodeling is pivotal for cell adhesion and migration. Here we investigated the function of Gas2-related protein on chromosome 22 (GAR22β), a poorly characterized protein that interacts with actin and microtubules. Primary and immortalized GAR22β-/- Sertoli cells moved faster than wild-type cells. In addition, GAR22β-/- cells showed a more prominent focal adhesion turnover. GAR22β overexpression or its reexpression in GAR22β-/- cells reduced cell motility and focal adhesion turnover. GAR22β-actin interaction was stronger than GAR22β-microtubule interaction, resulting in GAR22β localization and dynamics that mirrored those of the actin cytoskeleton. Mechanistically, GAR22β interacted with the regulator of microtubule dynamics end-binding protein 1 (EB1) via a novel noncanonical amino acid sequence, and this GAR22β-EB1 interaction was required for the ability of GAR22β to modulate cell motility. We found that GAR22β is highly expressed in mouse testes, and its absence resulted in reduced spermatozoa generation, lower actin levels in testes, and impaired motility and ultrastructural disorganization of spermatozoa. Collectively our findings identify GAR22β as a novel regulator of cell adhesion and migration and provide a foundation for understanding the molecular basis of diverse cytoskeleton-dependent processes

An Improved Search for the Neutron Electric Dipole Moment

A permanent electric dipole moment of fundamental spin-1/2 particles violates both parity (P) and time re- versal (T) symmetry, and hence, also charge-parity (CP) symmetry since there is no sign of CPT-violation. The search for a neutron electric dipole moment (nEDM) probes CP violation within and beyond the Stan- dard Model. The experiment, set up at the Paul Scherrer Institute (PSI), an improved, upgraded version of the apparatus which provided the current best experimental limit, dn < 2.9E-26 ecm (90% C.L.), by the RAL/Sussex/ILL collaboration: Baker et al., Phys. Rev. Lett. 97, 131801 (2006). In the next two years we aim to improve the sensitivity of the apparatus to sigma(dn) = 2.6E-27 ecm corresponding to an upper limit of dn < 5E-27 ecm (95% C.L.), in case for a null result. In parallel the collaboration works on the design of a new apparatus to further increase the sensitivity to sigma(dn) = 2.6E-28 ecm.Comment: APS Division for particles and fields, Conference Proceedings, Two figure

Membrane permeation of arginine-rich cell-penetrating peptides independent of transmembrane potential as a function of lipid composition and membrane fluidity

Cell-penetrating peptides (CPPs) are prominent delivery vehicles to confer cellular entry of (bio-) macromolecules. Internalization efficiency and uptake mechanism depend, next to the type of CPP and cargo, also on cell type. Direct penetration of the plasma membrane is the preferred route of entry as this circumvents endolysosomal sequestration. However, the molecular parameters underlying this import mechanism are still poorly defined. Here, we make use of the frequently used HeLa and HEK cell lines to address the role of lipid composition and membrane potential. In HeLa cells, at low concentrations, the CPP nona-arginine (R9) enters cells by endocytosis. Direct membrane penetration occurs only at high peptide concentrations through a mechanism involving activation of sphingomyelinase which converts sphingomyelin into ceramide. In HEK cells, by comparison, R9 enters the cytoplasm through direct membrane permeation already at low concentrations. This direct permeation is strongly reduced at room temperature and upon cholesterol depletion, indicating a complex dependence on membrane fluidity and microdomain organisation. Lipidomic analyses show that in comparison to HeLa cells HEK cells have an endogenously low sphingomyelin content. Interestingly, direct permeation in HEK cells and also in HeLa cells treated with exogenous sphingomyelinase is independent of membrane potential. Membrane potential is only required for induction of sphingomyelinase-dependent uptake which is then associated with a strong hyperpolarization of membrane potential as shown by whole-cell patch clamp recordings. Next to providing new insights into the interplay of membrane composition and direct permeation, these results also refute the long-standing paradigm that transmembrane potential is a driving force for CPP uptake

An Improved Neutron Electric Dipole Moment Experiment

A new measurement of the neutron EDM, using Ramsey's method of separated oscillatory fields, is in preparation at the new high intensity source of ultra-cold neutrons (UCN) at the Paul Scherrer Institute, Villigen, Switzerland (PSI). The existence of a non-zero nEDM would violate both parity and time reversal symmetry and, given the CPT theorem, might lead to a discovery of new CP violating mechanisms. Already the current upper limit for the nEDM (|d_n|<2.9E-26 e.cm) constrains some extensions of the Standard Model. The new experiment aims at a two orders of magnitude reduction of the experimental uncertainty, to be achieved mainly by (1) the higher UCN flux provided by the new PSI source, (2) better magnetic field control with improved magnetometry and (3) a double chamber configuration with opposite electric field directions. The first stage of the experiment will use an upgrade of the RAL/Sussex/ILL group's apparatus (which has produced the current best result) moved from Institut Laue-Langevin to PSI. The final accuracy will be achieved in a further step with a new spectrometer, presently in the design phase.Comment: Flavor Physics & CP Violation Conference, Taipei, 200

Gravitational depolarization of ultracold neutrons: comparison with data

We compare the expected effects of so-called gravitationally enhanced depolarization of ultracold neutrons to measurements carried out in a spin-precession chamber exposed to a variety of vertical magnetic-field gradients. In particular, we have investigated the dependence upon these field gradients of spin-depolarization rates and also of shifts in the measured neutron Larmor precession frequency. We find excellent qualitative agreement, with gravitationally enhanced depolarization accounting for several previously unexplained features in the data

Sensory Measurements: Coordination and Standardization

Do sensory measurements deserve the label of “measurement”? We argue that they do. They fit with an epistemological view of measurement held in current philosophy of science, and they face the same kinds of epistemological challenges as physical measurements do: the problem of coordination and the problem of standardization. These problems are addressed through the process of “epistemic iteration,” for all measurements. We also argue for distinguishing the problem of standardization from the problem of coordination. To exemplify our claims, we draw on olfactory performance tests, especially studies linking olfactory decline to neurodegenerative disorders

Cubic boron nitride: a new prospective material for ultracold neutron application

For the first time, the neutron optical wall-potential of natural cubic boron nitride (cBN) was measured at the ultracold neutron (UCN) source of the research reactor TRIGA Mainz using the time-of-flight method (TOF). The samples investigated had a wall-potential of (305 +/- 15) neV. This value is in good agreement with the result extracted from neutron reflectometry data and theoretical expectations. Because of its high critical velocity for UCN and its good dielectric characteristics, cubic boron nitride coatings (isotopically enriched) will be useful for a number of applications in UCN experiments

Chemotactic response and adaptation dynamics in Escherichia coli

Adaptation of the chemotaxis sensory pathway of the bacterium Escherichia coli is integral for detecting chemicals over a wide range of background concentrations, ultimately allowing cells to swim towards sources of attractant and away from repellents. Its biochemical mechanism based on methylation and demethylation of chemoreceptors has long been known. Despite the importance of adaptation for cell memory and behavior, the dynamics of adaptation are difficult to reconcile with current models of precise adaptation. Here, we follow time courses of signaling in response to concentration step changes of attractant using in vivo fluorescence resonance energy transfer measurements. Specifically, we use a condensed representation of adaptation time courses for efficient evaluation of different adaptation models. To quantitatively explain the data, we finally develop a dynamic model for signaling and adaptation based on the attractant flow in the experiment, signaling by cooperative receptor complexes, and multiple layers of feedback regulation for adaptation. We experimentally confirm the predicted effects of changing the enzyme-expression level and bypassing the negative feedback for demethylation. Our data analysis suggests significant imprecision in adaptation for large additions. Furthermore, our model predicts highly regulated, ultrafast adaptation in response to removal of attractant, which may be useful for fast reorientation of the cell and noise reduction in adaptation.Comment: accepted for publication in PLoS Computational Biology; manuscript (19 pages, 5 figures) and supplementary information; added additional clarification on alternative adaptation models in supplementary informatio

Measurement of the permanent electric dipole moment of the neutron

We present the result of an experiment to measure the electric dipole moment EDM) of the neutron at the Paul Scherrer Institute using Ramsey's method of separated oscillating magnetic fields with ultracold neutrons (UCN). Our measurement stands in the long history of EDM experiments probing physics violating time reversal invariance. The salient features of this experiment were the use of a Hg-199 co-magnetometer and an array of optically pumped cesium vapor magnetometers to cancel and correct for magnetic field changes. The statistical analysis was performed on blinded datasets by two separate groups while the estimation of systematic effects profited from an unprecedented knowledge of the magnetic field. The measured value of the neutron EDM is d_{\rm n} = (0.0\pm1.1_{\rm stat}\pm0.2_{\rmsys})\times10^{-26}e\,{\rm cm}