AMPA receptor anchoring at CA1 synapses is determined by N-terminal domain and TARP γ8 interactions.

AMPA receptor (AMPAR) abundance and positioning at excitatory synapses regulates the strength of transmission. Changes in AMPAR localisation can enact synaptic plasticity, allowing long-term information storage, and is therefore tightly controlled. Multiple mechanisms regulating AMPAR synaptic anchoring have been described, but with limited coherence or comparison between reports, our understanding of this process is unclear. Here, combining synaptic recordings from mouse hippocampal slices and super-resolution imaging in dissociated cultures, we compare the contributions of three AMPAR interaction domains controlling transmission at hippocampal CA1 synapses. We show that the AMPAR C-termini play only a modulatory role, whereas the extracellular N-terminal domain (NTD) and PDZ interactions of the auxiliary subunit TARP γ8 are both crucial, and each is sufficient to maintain transmission. Our data support a model in which γ8 accumulates AMPARs at the postsynaptic density, where the NTD further tunes their positioning. This interplay between cytosolic (TARP γ8) and synaptic cleft (NTD) interactions provides versatility to regulate synaptic transmission and plasticity

The gains and losses of face in ongoing intercultural interaction: A case study of Chinese participant perspectives

Given the small number of existing studies of face in intercultural settings and the increasing attention given to participant perspectives in face research, this paper explores the gains and losses of face as perceived by Chinese government officials during a three-week delegation visit to the United States of America. These perspectives were obtained from the group’s spontaneous discussions during regular evening meetings when they reflected on the day’s events. Several key features emerged from the discussions. Firstly, face enhancement was a primary goal for the visit – enhancement of their own face as a delegation, of the face of the Ministry they belonged to, as well as the face of their American hosts. Secondly, the delegates attempted to manage these face goals strategically. Thirdly, they spoke of face as a volatile image that could rise and fall sharply and yet endured across incidents, days and weeks. The paper reports on and discusses these participant perspectives in the light of recent theorizing on face

Chaotic properties of a turbulent isotropic fluid

By tracking the divergence of two initially close trajectories in phase space in an Eulerian approach to forced turbulence, the relation between the maximal Lyapunov exponent $\lambda$, and the Reynolds number $Re$ is measured using direct numerical simulations, performed on up to $2048^3$ collocation points. The Lyapunov exponent is found to solely depend on the Reynolds number with $\lambda \propto Re^{0.53}$ and that after a transient period the divergence of trajectories grows at the same rate at all scales. Finally a linear divergence is seen that is dependent on the energy forcing rate. Links are made with other chaotic systems.Comment: 8 pages, 8 figure

The uses of coherent structure (Dryden Lecture)

The concept of coherent structure in turbulent flow is a revolutionary idea which is being developed by evolutionary means. The main objective of this review is to list some solid achievements, showing what can be done by using the concept of coherent structure that cannot be done without it. The nature of structure is described in terms of some related concepts, including celerity, topology, and the phenomenon of coalescence and splitting of structure. The main emphasis is on the mixing layer, as the one flow whose structure is well enough understood so that technical applications are now being made in problems of mixing and chemistry. An attempt is made to identify some conceptual and experimental obstacles that stand in the way of progress in other technically important flows, particularly the turbulent boundary layer. A few comments are included about the role of structure in numerical simulations and in current work on manipulation and control of turbulent flow. Some recent developments are cited which suggest that the time is nearly right for corresponding advances to occur in turbulence modeling

Effects of wind speed changes on wake instability of a wind turbine in a virtual wind tunnel using large eddy simulation

Large Eddy Simulation (LES) of the National Renewable Energy Laboratory (NREL) Phase VI wind turbine inside a virtual wind tunnel, with the same test section as that of NASA Ames 24.4. m×36.6. m, was carried out in order to analyze and better understand the wake instability and its breakdown behind the wind turbine. LES was performed using the commercial CFD software, ANSYS FLUENT, based on the dynamic Smagorinsky-Lilly model. The wind turbine was placed at a distance of two rotor diameters from the upstream boundary with a downstream domain of 20 rotor diameters in length. The results of the simulation were compared with the experimental data published by the NREL and a good agreement was found between the two. Furthermore, the average turbulence intensities from the LES were compared with a semi-empirical model and very good agreement was observed, except for the regions of on-going wake instability and vortex breakdown. It was observed that the wake behind the wind turbine consists of a system of intense and stable rotating helical vortices. These vortices persisted for some distance downstream of the wind turbine and finally become unstable producing a sinuous shape. The downstream distance at which wake instability and vortex breakdown occur, was observed to be a function of the upstream wind speed. For example, for an upstream wind speed of 7. m/s, it was observed that the primary vortex structure became unstable at a downstream distance of four rotor diameters and complete breakdown occurred at approximately six rotor diameters. On the other hand, when the upstream wind speed was 15.1. m/s, wake instability occurred at approximately 11 rotor diameters downstream of the wind turbine and complete breakdown was observed at 13 rotor diameters downstream of the wind turbine. Furthermore, it was observed that the turbulence intensity rapidly decreased during the process of wake instability and vortex breakdown; the location of the decrease is a function of the upstream wind speed. It is suggested that the distinction between the near and far wake can be identified as the average location between the start of the wake instability and the end of the process, at complete breakdown. Therefore the average location of this boundary is a function of the upstream wind speed. Hence for upstream wind speeds of 7. m/s, 10. m/s, 13.1. m/s and 15.1. m/s, the boundary between the near and far wake lies at five, seven, ten and twelve rotor diameters downstream respectively. © 2013 Elsevier Ltd.Jang-Oh Mo, Amanullah Choudhry, Maziar Arjomandi, Richard Kelso, Young-Ho Le

A fully-automated home-cage for long-term continuous phenotyping of mouse cognition and behaviour

Automated home-cage monitoring systems present a valuable tool for comprehensive phenotyping of natural behaviours. However, current systems often involve complex training routines, water or food restriction, and probe a limited range of behaviours. Here, we present a fully automated home-cage monitoring system for cognitive and behavioural phenotyping in mice. The system incorporates T-maze-alternation, novel-object recognition and object-in-place recognition tests combined with monitoring of locomotion, drinking and quiescence patterns, carried out over long periods. Mice learn the tasks rapidly without any need for water or food restrictions. Behavioural characterization employs a deep convolutional neural network image analysis. We show that combined statistical properties of multiple behaviours can be used to discriminate between mice with hippocampal, medial entorhinal and sham lesions and predict the genotype of Alzheimer's disease mouse models with high accuracy. This technology may enable large-scale behavioural screening for genes and neural circuits underlying spatial memory and other cognitive processes.UK DR

Proton-triggered rearrangement of the AMPA receptor N-terminal domains impacts receptor kinetics and synaptic localization.

Acknowledgements: We thank C. Johnson for running the MALS experiment, N. Barry and J. Boulanger for helpful comments on FRAP imaging and analysis, R. Lape for providing the NSFA script and J.-N. Dohrke for helpful suggestions concerning the MD simulations. We also thank the Greger lab, J. Krieger and J. Watson for comments on the paper. We acknowledge the technical support provided by the Laboratory of Molecular Biology (LMB) Biophysics Facility, the Ares Biomedical Facility, LMB scientific computing and the LMB EM Facility. We acknowledge the use of the cryo-EM facility at the Center for Structural Biology (maintained by M. Chambers, S. Collier and M. Haider), ACCRE graphics processing unit nodes (supported by National Institutes of Health grant 1S10OD032234-01) and the Distributed Online Research Storage core at Vanderbilt University. We thank K. Kim and P. Christov at the Vanderbilt Chemical Synthesis Core for synthesizing chemicals. This work was supported by grants from the Medical Research Council (MC_U105174197), the Biotechnology and Biological Sciences Research Council (BB/N002113/1) and the Wellcome Trust (223194/Z/21/Z) to I.H.G. and NIH grants (R56/R01MH123474 and S10OD030292-01) to T.N.AMPA glutamate receptors (AMPARs) are ion channel tetramers that mediate the majority of fast excitatory synaptic transmission. They are composed of four subunits (GluA1-GluA4); the GluA2 subunit dominates AMPAR function throughout the forebrain. Its extracellular N-terminal domain (NTD) determines receptor localization at the synapse, ensuring reliable synaptic transmission and plasticity. This synaptic anchoring function requires a compact NTD tier, stabilized by a GluA2-specific NTD interface. Here we show that low pH conditions, which accompany synaptic activity, rupture this interface. All-atom molecular dynamics simulations reveal that protonation of an interfacial histidine residue (H208) centrally contributes to NTD rearrangement. Moreover, in stark contrast to their canonical compact arrangement at neutral pH, GluA2 cryo-electron microscopy structures exhibit a wide spectrum of NTD conformations under acidic conditions. We show that the consequences of this pH-dependent conformational control are twofold: rupture of the NTD tier slows recovery from desensitized states and increases receptor mobility at mouse hippocampal synapses. Therefore, a proton-triggered NTD switch will shape both AMPAR location and kinetics, thereby impacting synaptic signal transmission

Structural mobility tunes signalling of the GluA1 AMPA glutamate receptor

AMPA glutamate receptors (AMPARs), the primary mediators of excitatory neurotransmission in the brain, are either GluA2 subunit-containing and thus Ca2+-impermeable, or GluA2-lacking and Ca2+-permeable1. Despite their prominent expression throughout interneurons and glia, their role in long-term potentiation and their involvement in a range of neuropathologies2, structural information for GluA2-lacking receptors is currently absent. Here we determine and characterize cryo-electron microscopy structures of the GluA1 homotetramer, fully occupied with TARPγ3 auxiliary subunits (GluA1/γ3). The gating core of both resting and open-state GluA1/γ3 closely resembles GluA2-containing receptors. However, the sequence-diverse N-terminal domains (NTDs) give rise to a highly mobile assembly, enabling domain swapping and subunit re-alignments in the ligand-binding domain tier that are pronounced in desensitized states. These transitions underlie the unique kinetic properties of GluA1. A GluA2 mutant (F231A) increasing NTD dynamics phenocopies this behaviour, and exhibits reduced synaptic responses, reflecting the anchoring function of the AMPAR NTD at the synapse. Together, this work underscores how the subunit-diverse NTDs determine subunit arrangement, gating properties and ultimately synaptic signalling efficiency among AMPAR subtypes.Peer reviewe

Structural mobility tunes signalling of the GluA1 AMPA glutamate receptor

Acknowledgements: We thank G. Murshudov for suggestions for model building and S. Scheres for comments on EM processing. We also thank B. Singh and J. Watson for comments on the manuscript. We acknowledge S. Nayak for help with Fig. 4e design and L. Catapano for help with Coot images in Extended Data Figs. 4 and 6. We are grateful to LMB Scientific Computing and the EM Facility for support. This work was supported by grants from the Medical Research Council (grant no. MC_U105174197) and the Wellcome Trust (grant no. 223194/Z/21/Z) to I.H.G., and H2020 Marie Skłodowska-Curie Actions (grant no. 101024130) to J.M.K.AMPA glutamate receptors (AMPARs), the primary mediators of excitatory neurotransmission in the brain, are either GluA2 subunit-containing and thus Ca2+-impermeable, or GluA2-lacking and Ca2+-permeable1. Despite their prominent expression throughout interneurons and glia, their role in long-term potentiation and their involvement in a range of neuropathologies2, structural information for GluA2-lacking receptors is currently absent. Here we determine and characterize cryo-electron microscopy structures of the GluA1 homotetramer, fully occupied with TARPγ3 auxiliary subunits (GluA1/γ3). The gating core of both resting and open-state GluA1/γ3 closely resembles GluA2-containing receptors. However, the sequence-diverse N-terminal domains (NTDs) give rise to a highly mobile assembly, enabling domain swapping and subunit re-alignments in the ligand-binding domain tier that are pronounced in desensitized states. These transitions underlie the unique kinetic properties of GluA1. A GluA2 mutant (F231A) increasing NTD dynamics phenocopies this behaviour, and exhibits reduced synaptic responses, reflecting the anchoring function of the AMPAR NTD at the synapse. Together, this work underscores how the subunit-diverse NTDs determine subunit arrangement, gating properties and ultimately synaptic signalling efficiency among AMPAR subtypes