Structure of the human M2 muscarinic acetylcholine receptor bound to an antagonist

The parasympathetic branch of the autonomic nervous system regulates the activity of multiple organ systems. Muscarinic receptors are G-protein-coupled receptors that mediate the response to acetylcholine released from parasympathetic nerves. Their role in the unconscious regulation of organ and central nervous system function makes them potential therapeutic targets for a broad spectrum of diseases. The M2 muscarinic acetylcholine receptor (M2 receptor) is essential for the physiological control of cardiovascular function through activation of G-protein-coupled inwardly rectifying potassium channels, and is of particular interest because of its extensive pharmacological characterization with both orthosteric and allosteric ligands. Here we report the structure of the antagonist-bound human M2 receptor, the first human acetylcholine receptor to be characterized structurally, to our knowledge. The antagonist 3-quinuclidinyl-benzilate binds in the middle of a long aqueous channel extending approximately two-thirds through the membrane. The orthosteric binding pocket is formed by amino acids that are identical in all five muscarinic receptor subtypes, and shares structural homology with other functionally unrelated acetylcholine binding proteins from different species. A layer of tyrosine residues forms an aromatic cap restricting dissociation of the bound ligand. A binding site for allosteric ligands has been mapped to residues at the entrance to the binding pocket near this aromatic cap. The structure of the M2 receptor provides insights into the challenges of developing subtype-selective ligands for muscarinic receptors and their propensity for allosteric regulation.     

Palaeo-altimetry of the late Eocene to Miocene Lunpola basin, central Tibet

The elevation history of the Tibetan plateau provides direct insight into the tectonic processes associated with continent-continent collisions. Here we present oxygen-isotope-based estimates of the palaeo-altimetry of late Eocene and younger deposits of the Lunpola basin in the centre of the plateau, which indicate that the surface of Tibet has been at an elevation of more than 4 kilometres for at least the past 35 million years. We conclude that crustal, but not mantle, thickening models, combined with plate-kinematic solutions of India-Asia convergence, are compatible with palaeo-elevation estimates across the Tibetan plateau.     

Analysis of the DNA sequence and duplication history of human chromosome 15

Here we present a finished sequence of human chromosome 15, together with a high-quality gene catalogue. As chromosome 15 is one of seven human chromosomes with a high rate of segmental duplication, we have carried out a detailed analysis of the duplication structure of the chromosome. Segmental duplications in chromosome 15 are largely clustered in two regions, on proximal and distal 15q; the proximal region is notable because recombination among the segmental duplications can result in deletions causing Prader-Willi and Angelman syndromes. Sequence analysis shows that the proximal and distal regions of 15q share extensive ancient similarity. Using a simple approach, we have been able to reconstruct many of the events by which the current duplication structure arose. We find that most of the intrachromosomal duplications seem to share a common ancestry. Finally, we demonstrate that some remaining gaps in the genome sequence are probably due to structural polymorphisms between haplotypes; this may explain a significant fraction of the gaps remaining in the human genome.     

Planar cell polarity signalling couples cell division and morphogenesis during neurulation

Environmental and genetic aberrations lead to neural tube closure defects (NTDs) in 1 out of every 1,000 births. Mouse and frog models for these birth defects have indicated that Van Gogh-like 2 (Vangl2, also known as Strabismus) and other components of planar cell polarity (PCP) signalling might control neurulation by promoting the convergence of neural progenitors to the midline. Here we show a novel role for PCP signalling during neurulation in zebrafish. We demonstrate that non-canonical Wnt/PCP signalling polarizes neural progenitors along the anteroposterior axis. This polarity is transiently lost during cell division in the neural keel but is re-established as daughter cells reintegrate into the neuroepithelium. Loss of zebrafish Vangl2 (in trilobite mutants) abolishes the polarization of neural keel cells, disrupts re-intercalation of daughter cells into the neuroepithelium, and results in ectopic neural progenitor accumulations and NTDs. Remarkably, blocking cell division leads to rescue of trilobite neural tube morphogenesis despite persistent defects in convergence and extension. These results reveal a function for PCP signalling in coupling cell division and morphogenesis at neurulation and indicate a previously unrecognized mechanism that might underlie NTDs.     

Endothelial tubes assemble from intracellular vacuoles in vivo

The formation of epithelial tubes is crucial for the proper development of many different tissues and organs, and occurs by means of a variety of different mechanisms. Morphogenesis of seamless, properly patterned endothelial tubes is essential for the development of a functional vertebrate circulatory system, but the mechanism of vascular lumenization in vivo remains unclear. Evidence dating back more than 100 years has hinted at an important function for endothelial vacuoles in lumen formation. More than 25 years ago, in some of the first endothelial cell culture experiments in vitro, Folkman and Haudenschild described "longitudinal vacuoles" that "appeared to be extruded and connected from one cell to the next", observations confirmed and extended by later studies in vitro showing that intracellular vacuoles arise from integrin-dependent and cdc42/Rac1-dependent pinocytic events downstream of integrin-extracellular-matrix signalling interactions. Despite compelling data supporting a model for the assembly of endothelial tubes in vitro through the formation and fusion of vacuoles, conclusive evidence in vivo has been lacking, primarily because of difficulties associated with imaging the dynamics of subcellular endothelial vacuoles deep within living animals. Here we use high-resolution time-lapse two-photon imaging of transgenic zebrafish to examine how endothelial tubes assemble in vivo, comparing our results with time-lapse imaging of human endothelial-cell tube formation in three-dimensional collagen matrices in vitro. Our results provide strong support for a model in which the formation and intracellular and intercellular fusion of endothelial vacuoles drives vascular lumen formation.     

冷冻-解冻对烹饪后鸡胸肉的感官描述剖面的影响(英文)

冷冻是消费者用于延长肉类储藏期的一种常用方法,也是研究人员常用于肉类品质评估和加工的前道工序.目前尚无完善记录表明,冷冻对烹饪后的禽胸肉质的感官品质会产生影响.本研究的目的是比较新鲜与冷冻-解冻鸡胸肉的感官质量剖面属性.鸡胸肉块在宰后24 h内从胴体上分割,分别以新鲜状态直接烹饪或置于-20℃冷冻.冷冻样品采用三种不同解冻方式:冷冻后直接取出烹饪解冻(0 h);在20℃水中预解冻2 h后烹饪(2 h);或在4℃预解冻24 h后烹饪(24 h).对照组则为新鲜鸡胸肉直接烹饪.肉块加热到78℃时用于评价感官质量属性.该实验由经过训练的描述型品评员采用0~15强度标度法进行评价.结果显示:不同的处理方式并没有引起任何风味特征和5种质构特征(内聚力、硬度、多汁、肉块大小、肉块湿度)上的差异(P0.05).然而,不同处理方式引起了团块内聚性、分解率和咀嚼性上的显著差异(P0.05):冻后直接烹饪(0 h)和解冻24 h烹饪(24 h)的鸡肉块的团块内聚性评价要明显高于预解冻2 h的样品.冻后直接烹饪样品(0h)的分解率和咀嚼性显著高于新鲜对照组和预解冻2 h的样品.本研究结果表明,冷冻-解冻对鸡胸肉的感官风味品质无影响,但可能改变烹饪后鸡胸肉的质构特征.因此,冷冻-解冻对肉类质构的影响取决于烹饪前的解冻方法.