The formation of acetylcholine receptor clusters visualized with quantum dots

Background: Motor innervation of skeletal muscle leads to the assembly of acetylcholine receptor (AChR) clusters in the postsynaptic membrane at the vertebrate neuromuscular junction (NMJ). Synaptic AChR aggregation, according to the diffusion-mediated trapping hypothesis, involves the establishment of a postsynaptic scaffold that "traps" freely diffusing receptors into forming high-density clusters. Although this hypothesis is widely cited to explain the formation of postsynaptic AChR clusters, direct evidence at molecular level is lacking. Results: Using quantum dots (QDs) and live cell imaging, we provide new measurements supporting the diffusion-trap hypothesis as applied to AChR cluster formation. Consistent with published works, experiments on cultured Xenopus myotomal muscle cells revealed that AChRs at clusters that formed spontaneously (pre-patterned clusters, also called hot spots) and at those induced by nerve-innervation or by growth factor-coated latex beads were very stable whereas diffuse receptors outside these regions were mobile. Moreover, despite the restriction of AChR movement at sites of synaptogenic stimulation, individual receptors away from these domains continued to exhibit free diffusion, indicating that AChR clustering at NMJ does not involve an active attraction of receptors but is passive and diffusion-driven. Conclusion: Single-molecular tracking using QDs has provided direct evidence that the clustering of AChRs in muscle cells in response to synaptogenic stimuli is achieved by two distinct cellular processes: the Brownian motion of receptors in the membrane and their trapping and immobilization at the synaptic specialization. This study also provides a clearer picture of the "trap" that it is not a uniformly sticky area but consists of discrete foci at which AChRs are immobilized

Crosslinking-induced endocytosis of acetylcholine receptors by quantum dots

In a majority of patients with myasthenia gravis (MG), anti-acetylcholine receptor (AChR) antibodies target postsynaptic AChR clusters and thus compromise the membrane integrity of neuromuscular junctions (NMJs) and lead to muscle weakness. Antibody-induced endocytosis of AChRs in the postsynaptic membrane represents the initial step in the pathogenesis of MG; however, the molecular mechanisms underlying AChR endocytosis remain largely unknown. Here, we developed an approach to mimic the pathogenic antibodies for inducing the crosslinking and internalization of AChRs from the postsynaptic membrane. Using biotin-α-bungarotoxin and quantum dot (QD)-streptavidin, cell-surface and internalized AChRs could be readily distinguished by comparing the size, fluorescence intensity, trajectory, and subcellular localization of the QD signals. QD-induced AChR endocytosis was mediated by clathrin-dependent and caveolin-independent mechanisms, and the trafficking of internalized AChRs in the early endosomes required the integrity of microtubule structures. Furthermore, activation of the agrin/MuSK (muscle-specific kinase) signaling pathway strongly suppressed QD-induced internalization of AChRs. Lastly, QD-induced AChR crosslinking potentiated the dispersal of aneural AChR clusters upon synaptic induction. Taken together, our results identify a novel approach to study the mechanisms of AChR trafficking upon receptor crosslinking and endocytosis, and demonstrate that agrin-MuSK signaling pathways protect against crosslinking-induced endocytosis of AChRs. © 2014 Lee et al.published_or_final_versio

Effects of profile variability on aerodynamic performance of supersonic compressor cascade

ObjectivesThis study seeks to evaluate the effects of profile variability on the aerodynamic per-formance of a compressor and provide guidance for the robust design of compressor blades. MethodsA mathematical model of profile variability distribution with a single peak is established. The effects of the combined profile variability of the blade pressure and suction surface on the aerodynamic performance of two supersonic planar cascades are then investigated by numerical simulation. ResultsThe results show that the profile variability distribution on the suction surface is the key factor behind cascade total pressure loss. The total pressure loss coefficient decreases gradually with the position of maximum profile variability on the suction surface moving downstream. The profile variability distribution on the blade pressure and suction surface influences the flow turning angle and static pressure rise coefficient with opposite trends. The profile variability on the suction surface plays a dominant role in the flow turning angle and static pressure rise of cascade with lower incoming Mach number; for cascade with higher incoming Mach number, the profile variability on the pressure surface has a significant impact on the flow turning angle and static pressure rise. The position and intensity of the shockwave and the end wall profile of the expansion channel after the shockwave comprehensively determine the flow state on the blade surface and in the cascade blade passage. The flow loss near the blade suction surface increases, the flow loss near the blade pressure surface decreases, and the compound effect determines the change of cascade loss, flow turning angle and static pressure rise. ConclusionsThe results of this study can provide guidance for the design, manufacture and manufacturing variability evaluation of transonic compressors

Nanoindentation induced anisotropy of deformation and damage behaviors of MgF2 crystals

The competition mechanism between the slip motions and cleavage fractures is related to the anisotropy of deformation behaviors, which is essential to manufacture complex optical components. To identify competition mechanism between the slip motions and cleavage fractures and reveal the anisotropy of deformation and damage behaviors of MgF2 crystals, the nanoindentation tests were systematically conducted on different crystal planes. In addition, the stress induced by the nanoindentation was developed and decomposed along the slip systems and cleavage planes, and cleavage factors and Schmid factors were calculated. The stress, cleavage factors and Schmid factors indicated that the activation degree of the slip motions and cleavage fractures determined the indentation morphologies. Under the same indentation conditions, the nanoindentation of the (001) crystal plane activated most slip motions, so the plastic deformation is most prone to occur on this crystal plane. The nanoindentation of the (010) crystal plane activated less slip motions and most cleavage fractures, resulting in the severest brittle fractures on the (010) crystal plane. The theoretical results consisted well with the experimental results, which provides the theoretical guidance to the low-damage manufacturing of MgF2 components

The formation of acetylcholine receptor clusters visualized with quantum dots

Abstract Background Motor innervation of skeletal muscle leads to the assembly of acetylcholine receptor (AChR) clusters in the postsynaptic membrane at the vertebrate neuromuscular junction (NMJ). Synaptic AChR aggregation, according to the diffusion-mediated trapping hypothesis, involves the establishment of a postsynaptic scaffold that "traps" freely diffusing receptors into forming high-density clusters. Although this hypothesis is widely cited to explain the formation of postsynaptic AChR clusters, direct evidence at molecular level is lacking. Results Using quantum dots (QDs) and live cell imaging, we provide new measurements supporting the diffusion-trap hypothesis as applied to AChR cluster formation. Consistent with published works, experiments on cultured Xenopus myotomal muscle cells revealed that AChRs at clusters that formed spontaneously (pre-patterned clusters, also called hot spots) and at those induced by nerve-innervation or by growth factor-coated latex beads were very stable whereas diffuse receptors outside these regions were mobile. Moreover, despite the restriction of AChR movement at sites of synaptogenic stimulation, individual receptors away from these domains continued to exhibit free diffusion, indicating that AChR clustering at NMJ does not involve an active attraction of receptors but is passive and diffusion-driven. Conclusion Single-molecular tracking using QDs has provided direct evidence that the clustering of AChRs in muscle cells in response to synaptogenic stimuli is achieved by two distinct cellular processes: the Brownian motion of receptors in the membrane and their trapping and immobilization at the synaptic specialization. This study also provides a clearer picture of the "trap" that it is not a uniformly sticky area but consists of discrete foci at which AChRs are immobilized.</p

Quantifying Seepage‐Face Evaporation and Its Effects on Groundwater Flow and Solute Transport in Small‐Slope Tidal Flat

Abstract Large‐scale seepage faces occur on tidal flats with gentle slope, which are widely distributed worldwide. Evaporation on these seepage faces, leading to salt retention and accumulation, may significantly impact the density‐dependent groundwater flow beneath the tidal flats. However, due to nonlinear complexities of the groundwater flow and solute transport on seepage faces, explicit boundary conditions and numerical models to quantify these processes are lacking. In this study, we present both mathematical and numerical models to quantify these processes. Compared to the results of our previous study, this paper shows that seepage‐face evaporation can (a) significantly increase the groundwater salinity in the upper intertidal zone, and form multiple groundwater circulation cells in the intertidal zone, (b) cause the disappearance of multiple seepage‐faces and reduce the spatial extent of seepage faces notably, (c) and intensify the groundwater and salt exchange as well as the seawater‐groundwater circulation through the intertidal zone

Characterization of the complete chloroplast genome of medicinal tea tree (Melaleuca alternifolia)

Melaleuca alternifolia is commonly known as the medicinal tea tree. The complete chloroplast (cp) genome sequence is 160,104 bp in length, with a quantitative molecule structure comprising two copies of inverted repeats (IRa and IRb) of 26,737 bp separated by a large single copy (LSC) of 88,151bp, a small single copy (SSC) of 18,479 bp. A total of 131 genes were identified including 84 protein-coding genes, 37 tRNA genes, eight rRNA genes and two pseudogene (Ψycf1, ΨinfA), respectively. Phylogenomic analysis suggests that M. alternifolia is closely related to the rest species of Myrtaceae with strong bootstrap values