The tight junction does not allow lipid molecules to diffuse from one epithelial cell to the next

The tight junction (zonula occludens) links epithelial cells into a monolayer by forming a continuous belt of sealing contacts around the apex of each cell. They appear in thin sections as if they were 'fusions' between the apposed plasma membranes and in freeze-fracture replicas as patterns of complementary strands and furrows. These images have led to the proposal that the core of the tight junction is formed by a hexagonal cylinder of lipids. In this model, the cytoplasmic leaflet of the apical and basolateral plasma membrane domains would be continuous, whereas the exoplasmic leaflets of the two plasma membrane domains of the same cell would be separated at the tight junction and are instead predicted to be continuous between the plasma membranes of neighbouring cells. We demonstrate here that this prediction does not hold true. An endogenous glycolipid (Forssman antigen), present in the exoplasmic leaflet of the apical membrane of MDCK strain II cells, is unable to pass to MDCK strain I cells (which lack this glycolipid) under conditions where these cells are connected by tight junctions. In addition, fluorescent lipids which have been fused into the plasma membrane of one MDCK cell do not diffuse to neighbouring cells while the tight junctions between the cells are intact.     

Homologies between gap junction proteins in lens, heart and liver

The cells in the mammalian lens are electrically and metabolically coupled with each other by a network of gap junctions. These are clusters of transmembrane channels by which the fibre cells situated deeper in the lens communicate through the epithelium with the aqueous humour, the source of nutrients for the lens. Hence gap junctions are important for lens transparency. The gap junction proteins in the mammalian lens have not yet been identified with certainty. A putative fibre gap junction protein of relative molecular mass 26,000 (26K) is not related to those from other tissues, such as the liver 28K junction component. Another lens membrane protein with Mr 70K (MP70) has also been localized in the lens fibre gap junctions. Here we demonstrate by amino-terminal sequence analysis that MP70 and its in vivo-processed form, MP38 (ref. 8), belong to a wider family of gap junction proteins. With this new data on the lens, homologies between gap junction proteins now extend to organs derived from all three embryonal layers, endoderm (liver), mesoderm (heart) and ectoderm (lens).     

间隙连接在鸡胚水晶体发育中的作用

初步报道已揭示，单克隆抗体ＮＤ６可影响鸡胚水晶体的发育，使之明显增大．ＮＤ６是一种被认为专一于膜蛋白ＭＰ２６细胞外侧段的单抗．ＭＰ２６已被认为是水晶体纤维细胞间隙连接的成份．因此，ＮＤ６很可能可阻断水晶体纤维细胞间隙连接的形成．本人试图用定量显微镜技术证实ＮＤ６对水晶体发育的影响与间隙连接数量减少之间的相关性。注射ＮＤ６于２０期鸡胚的右眼；同胚未注射的左眼作为对照．培养２４ｈ后周定，然后作下列三种处理：（１）测量整体水晶体的大小；（２）制备水晶体超薄切片，并统计间隙连接数量；（３）制备鸡胚头部的连续石蜡切片，并统计水晶体纤维细胞的数量．实验结果指出，在ＮＤ６处理２４ｈ，水晶体大小及纤维细胞数量均比对照组明显增大（表１，Ｐ＜０．０１或０．００１）；而间隙连接数量则比对照组明显减少（表２），Ｐ＜０．００１）．这表明，水晶体的增大及纤维细胞的增多是由间隙连接的减少所引起．这些结果证实，间隙连接在鸡胚水晶体的发育中起重要作用．     

Centrifugal fibres synapse on dopaminergic interplexiform cells in the teleost retina

In teleost fish, centrifugal fibres originating in the olfactory bulb and containing FMRFamide-like and luteinizing hormone releasing hormone (LHRH)-like peptides project to the retina and terminate along the border of the inner nuclear and inner plexiform layers. Using a novel simultaneous two-colour immunolabelling technique, we have found that these centrifugal fibres are often closely apposed to the dopaminergic interplexiform cells. Contacts between centrifugal fibres and dopaminergic interplexiform cells were observed by electron microscopy to be conventional type synaptic junctions. Since the dopaminergic interplexiform cells make synapses on horizontal and bipolar cells, providing an intraretinal centrifugal pathway for information flow from the inner to the outer plexiform layers, we conclude that every neuron in the teleost retina is potentially susceptible to central influences via these centrifugal fibres and dopaminergic interplexiform cells.     

Planar polarized actomyosin contractile flows control epithelial junction remodelling

Force generation by Myosin-II motors on actin filaments drives cell and tissue morphogenesis. In epithelia, contractile forces are resisted at apical junctions by adhesive forces dependent on E-cadherin, which also transmits tension. During Drosophila embryonic germband extension, tissue elongation is driven by cell intercalation, which requires an irreversible and planar polarized remodelling of epithelial cell junctions. We investigate how cell deformations emerge from the interplay between force generation and cortical force transmission during this remodelling in Drosophila melanogaster. The shrinkage of dorsal-ventral-oriented ('vertical') junctions during this process is known to require planar polarized junctional contractility by Myosin II (refs 4, 5, 7, 12). Here we show that this shrinkage is not produced by junctional Myosin II itself, but by the polarized flow of medial actomyosin pulses towards 'vertical' junctions. This anisotropic flow is oriented by the planar polarized distribution of E-cadherin complexes, in that medial Myosin II flows towards 'vertical' junctions, which have relatively less E-cadherin than transverse junctions. Our evidence suggests that the medial flow pattern reflects equilibrium properties of force transmission and coupling to E-cadherin by α-Catenin. Thus, epithelial morphogenesis is not properly reflected by Myosin II steady state distribution but by polarized contractile actomyosin flows that emerge from interactions between E-cadherin and actomyosin networks.     

Cingulin, a new peripheral component of tight junctions

The tight junction (Zonula occludens), a belt-like region of contact between cells of polarized epithelia, serves as a selective barrier to small molecules and as a total barrier to large molecules, and is involved in the separation between lumenal and basolateral compartments of the epithelium. In the electron microscope, tight junctions show focal regions of apparent fusion between the adjoining cell membranes, and freeze-fractured membranes display an elaborate network of branching and anastomosing strands. Very little is known about the molecular composition and architecture of tight junctions. The first specific zonula occludens-associated protein, designated ZO-1, has recently been identified in mammalian epithelial and endothelial cells. Here we describe the identification and purification of a new component of this junctional complex in avian brush-border cells, which we name cingulin. Cingulin is an acidic, heat-stable protein, with a highly elongated shape. Immunofluorescence and immunoelectron microscopy of brush-border cells with anti-cingulin antibodies show that cingulin is localized in the apical zone of the terminal web, at the endofacial surfaces of the zonula occludens.     

Gap junction adhesion is necessary for radial migration in the neocortex

Radial glia, the neuronal stem cells of the embryonic cerebral cortex, reside deep within the developing brain and extend radial fibres to the pial surface, along which embryonic neurons migrate to reach the cortical plate. Here we show that the gap junction subunits connexin 26 (Cx26) and connexin 43 (Cx43) are expressed at the contact points between radial fibres and migrating neurons, and acute downregulation of Cx26 or Cx43 impairs the migration of neurons to the cortical plate. Unexpectedly, gap junctions do not mediate neuronal migration by acting in the classical manner to provide an aqueous channel for cell-cell communication. Instead, gap junctions provide dynamic adhesive contacts that interact with the internal cytoskeleton to enable leading process stabilization along radial fibres as well as the subsequent translocation of the nucleus. These results indicate that gap junction adhesions are necessary for glial-guided neuronal migration, raising the possibility that the adhesive properties of gap junctions may have an important role in other physiological processes and diseases associated with gap junction function.     

Patterns of dye coupling in the imaginal wing disk of Drosophila melanogaster

Responses of developing tissues to experimental disruption demonstrate that cell interaction is important both in generating positional information and in controlling growth. However, the mechanism by which cells interact and the range over which the interactions are effective are not known. In the imaginal disks of Drosophila melanogaster, experiments on pattern regulation following surgical ablation suggest that the cell interactions are very local in nature; in fact, most of the data can be explained by assuming that cells interact only with their immediate neighbours. In contrast, studies of cell division patterns in the same tissue indicate that the "local' proliferative response to an ablation extends over a distance of up to about eight cell diameters. Still longer-range interactions have been proposed on the basis of theoretical considerations. It is possible that the interactions are mediated by the transfer of small molecules through gap junctions, as gap junctions are abundant in imaginal disks at the appropriate developmental stages. We have explored the range, timing and directionality of dye coupling between the cells of the wing disk as a test of the possible role of gap junctions in imaginal disk patterning. Our results indicate that interactions over different ranges are possible depending on the nature of the molecule being transferred.     

Patterns of junctional communication in the early amphibian embryo

It has long been recognized that cells in early embryos can communicate with each other via a direct cell-to-cell pathway, probably mediated by gap junctions. Low electrical resistance pathways, detected electrophysiologically, have been identified in all species examined so far. However, studies in various embryos on the transfer of molecules larger than small ions (for example, fluorescent dyes in the molecular weight range 350-500) have given conflicting results. In all these studies the ability to transfer dyes from cell to cell was determined without reference to the position of the injected cell in the embryo. In the experiments reported here, cell-cell transfer of the fluorescent dye, Lucifer yellow (molecular weight (Mr) 450) was re-examined in the early Xenopus laevis embryo by injecting the dye into identified cells, as the position of the injected cell within the embryo may be important. At the 32-cell stage, we found that dye transfer often occurred between animal pole blastomeres which were not sisters, as well as between sister cells, and also that Lucifer yellow was indeed transferred via gap junctions. The cell-cell transfer was not uniform within the animal pole; transfer was maximal near the dorsal side and minimal at the ventral side. This pattern may reflect differences in permeability or numbers of gap junctions across the embryo, and could be related to early events in development.     

Coherent transfer of Cooper pairs by a movable grain

Superconducting circuits that incorporate Josephson junctions are of considerable experimental and theoretical interest, particularly in the context of quantum computing. A nanometre-sized superconducting grain (commonly referred to as a Cooper-pair box) connected to a reservoir by a Josephson junction is an important example of such a system. Although the grain contains a large number of electrons, it has been experimentally demonstrated that its states are given by a superposition of only two charge states (differing by 2e, where e is the electronic charge). Coupling between charge transfer and mechanical motion in nanometre-sized structures has also received considerable attention. Here we demonstrate theoretically that a movable Cooper-pair box oscillating periodically between two remote superconducting electrodes can serve as a mediator of Josephson coupling, leading to coherent transfer of Cooper pairs between the electrodes. Both the magnitude and the direction of the resulting Josephson current can be controlled by externally applied electrostatic fields.     

深度优先局部聚合哈希

已有的深度监督哈希方法不能有效地利用提取到的卷积特征,同时,也忽视了数据对之间相似性信息分布对于哈希网络的作用,最终导致学到的哈希编码之间的区分性不足.为了解决该问题,提出了一种新颖的深度监督哈希方法,称之为深度优先局部聚合哈希(Deep Priority Local Aggregated Hashing,DPLAH).DPLAH将局部聚合描述子向量嵌入到哈希网络中,提高网络对同类数据的表达能力,并且通过在数据对之间施加不同权重,从而减少相似性信息分布倾斜对哈希网络的影响.利用Pytorch深度框架进行DPLAH实验,使用NetVLAD层对Resnet18网络模型输出的卷积特征进行聚合,将聚合得到的特征进行哈希编码学习.在CI-FAR-10和NUS-WIDE数据集上的图像检索实验表明,与使用手工特征和卷积神经网络特征的非深度哈希学习算法的最好结果相比,DPLAH的平均准确率均值要高出11％,同时,DPLAH的平均准确率均值比非对称深度监督哈希方法高出2％.     

Two homologous protein components of hepatic gap junctions

Gap junctions consist of closely packed pairs of transmembrane channels, the connexons, through which materials of low relative molecular mass diffuse from the cell to neighbouring cells. In liver, connexons consist of six protein subunits which, until now, were believed to be identical. However, besides the major polypeptide of relative molecular mass (Mr) 28,000 (and see refs 4 and 6), a component of Mr 21,000 (21K) has been repeatedly observed in liver. The amino-terminal sequence (18 residues) of this less abundant protein shows that it is related to, but distinct from, the Mr 28K protein. Immuno-staining and immuno-precipitation show both proteins to be in the same gap junctional plaques. Thus, it seems that hepatic gap junction channels (and by extension possibly others) are composed of two (or more) homologous proteins.     

Fluctuating superconductivity in organic molecular metals close to the Mott transition

On cooling through the transition temperature T(c) of a conventional superconductor, an energy gap develops as the normal-state charge carriers form Cooper pairs; these pairs form a phase-coherent condensate that exhibits the well-known signatures of superconductivity: zero resistivity and the expulsion of magnetic flux (the Meissner effect). However, in many unconventional superconductors, the formation of the energy gap is not coincident with the formation of the phase-coherent superfluid. Instead, at temperatures above the critical temperature a range of unusual properties, collectively known as 'pseudogap phenomena', are observed. Here we argue that a key pseudogap phenomenon-fluctuating superconductivity occurring substantially above the transition temperature-could be induced by the proximity of a Mott-insulating state. The Mott-insulating state in the kappa-(BEDT-TTF)2X organic molecular metals can be tuned, without doping, through superconductivity into a normal metallic state as a function of the parameter t/U, where t is the tight-binding transfer integral characterizing the metallic bandwidth and U is the on-site Coulomb repulsion. By exploiting a particularly sensitive probe of superconducting fluctuations, the vortex-Nernst effect, we find that a fluctuating regime develops as t/U decreases and the role of Coulomb correlations increases.     

Crystal structure of T4 endonuclease VII resolving a Holliday junction

Holliday proposed a four-way DNA junction as an intermediate in homologous recombination, and such Holliday junctions have since been identified as a central component in DNA recombination and repair. Phage T4 endonuclease VII (endo VII) was the first enzyme shown to resolve Holliday junctions into duplex DNAs by introducing symmetrical nicks in equivalent strands. Several Holliday junction resolvases have since been characterized, but an atomic structure of a resolvase complex with a Holliday junction remained elusive. Here we report the crystal structure of an inactive T4 endo VII(N62D) complexed with an immobile four-way junction with alternating arm lengths of 10 and 14 base pairs. The junction is a hybrid of the conventional square-planar and stacked-X conformation. Endo VII protrudes into the junction point from the minor groove side, opening it to a 14 A x 32 A parallelogram. This interaction interrupts the coaxial stacking, yet every base pair surrounding the junction remains intact. Additional interactions involve the positively charged protein and DNA phosphate backbones. Each scissile phosphate that is two base pairs from the crossover interacts with a Mg2+ ion in the active site. The similar overall shape and surface charge potential of the Holliday junction resolvases endo VII, RuvC, Ydc2, Hjc and RecU, despite having different folds, active site composition and DNA sequence preference, suggest a conserved binding mode for Holliday junctions.     

Cross-presentation by intercellular peptide transfer through gap junctions

Major histocompatibility complex (MHC) class I molecules present peptides that are derived from endogenous proteins. These antigens can also be transferred to professional antigen-presenting cells in a process called cross-presentation, which precedes initiation of a proper T-cell response; but exactly how they do this is unclear. We tested whether peptides can be transferred directly from the cytoplasm of one cell into the cytoplasm of its neighbour through gap junctions. Here we show that peptides with a relative molecular mass of up to approximately 1,800 diffuse intercellularly through gap junctions unless a three-dimensional structure is imposed. This intercellular peptide transfer causes cytotoxic T-cell recognition of adjacent, innocent bystander cells as well as activated monocytes. Gap-junction-mediated peptide transfer is restricted to a few coupling cells owing to the high cytosolic peptidase activity. We present a mechanism of antigen acquisition for cross-presentation that couples the antigen presentation system of two adjacent cells and is lost in most tumours: gap-junction-mediated intercellular peptide coupling for presentation by bystander MHC class I molecules and transfer to professional antigen presenting cells for cross-priming.     

Aquaporin-0 membrane junctions reveal the structure of a closed water pore

The lens-specific water pore aquaporin-0 (AQP0) is the only aquaporin known to form membrane junctions in vivo. We show here that AQP0 from the lens core, containing some carboxy-terminally cleaved AQP0, forms double-layered crystals that recapitulate in vivo junctions. We present the structure of the AQP0 membrane junction as determined by electron crystallography. The junction is formed by three localized interactions between AQP0 molecules in adjoining membranes, mainly mediated by proline residues conserved in AQP0s from different species but not present in most other aquaporins. Whereas all previously determined aquaporin structures show the pore in an open conformation, the water pore is closed in AQP0 junctions. The water pathway in AQP0 also contains an additional pore constriction, not seen in other known aquaporin structures, which may be responsible for pore gating.     

Emergence of preformed Cooper pairs from the doped Mott insulating state in Bi2Sr2CaCu2O8+delta

Superconductors are characterized by an energy gap that represents the energy needed to break the pairs of electrons (Cooper pairs) apart. At temperatures considerably above those associated with superconductivity, the high-transition-temperature copper oxides have an additional 'pseudogap'. It has been unclear whether this represents preformed pairs of electrons that have not achieved the coherence necessary for superconductivity, or whether it reflects some alternative ground state that competes with superconductivity. Paired electrons should display particle-hole symmetry with respect to the Fermi level (the energy of the highest occupied level in the electronic system), but competing states need not show such symmetry. Here we report a photoemission study of the underdoped copper oxide Bi(2)Sr(2)CaCu(2)O(8+delta) that shows the opening of a symmetric gap only in the anti-nodal region, contrary to the expectation that pairing would take place in the nodal region. It is therefore evident that the pseudogap does reflect the formation of preformed pairs of electrons and that the pairing occurs only in well-defined directions of the underlying lattice.     

物流管道内表面磁力研磨的数值仿真

由于物流管道内表面磁力研磨的特殊性，管道及励磁机构都不宜作高速旋转运动．提出了用电磁励磁产生回转磁场实现非机械方式产生相对运动的方法，并对励磁机构以简化模型为基础进行了数值仿真，使研磨压力、磁驱动力和简化模型的相关参数建立了数值上的关联，同时对数值计算的结果进行了具体的分析．研究结果表明：磁驱动力随间隙及相对位移增大都是先上升后下降，在大间隙区域内该力随相对位移变化趋缓；研磨压力随相对位移单边下降，在小间隙区域对研磨压力缺乏表达能力．     

信号控制交叉口左转相位协调设计方法

基于左转交通流的路径分析,给出了相邻交叉口左转相位协调优化的约束条件,包括流量守恒、绿灯最大最小时间、饱和度和周期时长等.考虑不同左转相位设置模式,建立了相邻交叉口间通行能力差计算模型.在此基础上,以通行能力差最小为目标,提出了相邻交叉口左转相位协调设计模型.最后,基于两个实际交叉口的几何条件和交通流数据,采用车均延误和最大通过量为指标对比分析了交叉口现有方案、Synchro优化方案和本文模型优化方案的信号控制效益.结果表明,基于本文模型的相位方案,能降低各交叉口间的通行能力差值,并有效降低交叉口群的车均延误,提高交叉口群整体通行能力,同时该协调设计方法对整个系统的负面影响较小.     

齿轮齿条式作动器性能分析与试验

为提高车辆悬架系统的减振性能,设计了一种用于车辆主动悬架系统的齿轮齿条式作动器。首先理论分析了作动器响应时间的影响因素,然后推导了作动器主动控制力的计算方法,最后通过台架试验进行了验证。试验结果表明:作动器主动控制力的实际值与理论值之间偏差较小;且与电机的供电电流之间呈现较好的线性关系,与理论分析一致;电流变化的响应时间基本相同;但作动器主动出力的响应时间随电流的减小而增大,分析认为这是由作动器的空程间隙造成的。因此,在作动器的设计过程中应尽量避免或减小空程间隙。