TFG clusters COPII‐coated transport carriers and promotes early secretory pathway organization

In mammalian cells, cargo‐laden secretory vesicles leave the endoplasmic reticulum (ER) en route to ER‐Golgi intermediate compartments (ERGIC) in a manner dependent on the COPII coat complex. We report here that COPII‐coated transport carriers traverse a submicron, TFG (Trk‐fused gene)‐enriched zone at the ER/ERGIC interface. The architecture of TFG complexes as determined by three‐dimensional electron microscopy reveals the formation of flexible, octameric cup‐like structures, which are able to self‐associate to generate larger polymers in vitro. In cells, loss of TFG function dramatically slows protein export from the ER and results in the accumulation of COPII‐coated carriers throughout the cytoplasm. Additionally, the tight association between ER and ERGIC membranes is lost in the absence of TFG. We propose that TFG functions at the ER/ERGIC interface to locally concentrate COPII‐coated transport carriers and link exit sites on the ER to ERGIC membranes. Our findings provide a new mechanism by which COPII‐coated carriers are retained near their site of formation to facilitate rapid fusion with neighboring ERGIC membranes upon uncoating, thereby promoting interorganellar cargo transport.     

Tarsal taste sensilla of the autumn gum moth, Mnesampela privata: morphology and electrophysiological activity

Mnesampela privata Guenée (Lepidoptera: Geometridae: Ennominae) is a native Australian geometrid that conducts considerable host assessment prior to ovipositing on its host plants, which belong to the genus Eucalyptus . The leaves of some of their hosts are covered with a particularly thick and waxy cuticle and we have shown that epicuticular waxes influence the oviposition preferences of females. This necessitates that M. privata has evolved specific chemosensory organs to assess the identity and perhaps even the quality of its hosts. In this work, we examined the morphology of tarsal taste sensilla and the sensitivity of their sensory neurones to a range of primary metabolites possibly influential on host assessment and oviposition. The ventral surface of the fifth tarsomere of females bear two parallel rows of up to eight sensilla, each loosely aligned with two parallel rows of five spines. Salts, sugars, and amino acids elicited phasi-tonic multicellular neuronal responses of variable magnitude and form. Two pairs of sensilla are closely apposed to the most distal spine in each row; the sensory neurones associated with these sensilla exhibited notably larger responses to alanine and serine compared with those of all other sensilla. The arrangement of the taste sensilla in close proximity to prominent tarsal spines is unique and could represent an adaptation that enables them to penetrate the wax layer and be brought into contact with primary metabolites present closer to the leaf surface.     

Branched-chain constituents of Brussels sprout wax

The presence of a substantial proportion of branched, in addition to normal compounds, was confirmed in the alkyl ester and primary alcohol components     

Chemotaxonomical relationships between penguins and tubenoses

The compositions of the uropygial gland wax of seven species of birds of the order Procellariiformes (tubenoses) belonging to four different families are compared with one another and with those of birds belonging to other orders. The results are discussed from a chemotaxonomical viewpoint. For this purpose, the various parameters have been depicted on a 3-dimensional matrix. Although all birds investigated are obviously related to each other, two of the species (Diomedea and Garrodia) also show a relationship to different orders. The results support the opinion that the Procellariiformes are closely related to the Sphenisciformes (penguins).     

Effect of proglumide, a cholecystokinin receptor antagonist, on caerulein-stimulated pancreatic enzyme secretion and pancreatic polypeptide release in the dog

In five conscious dogs we studied the effect of proglumide, a cholecystokinin (CCK) antagonist, on caerulein-stimulated pancreatic secretion and release of pancreatic polypeptide (PP). Graded doses of caerulein (15-240 ng/kg per h) were infused intravenously. Experiments were repeated with a fixed infusion of proglumide (40 mg/kg per h). Release of PP following increasing doses of caerulein was significantly inhibited by proglumide (P less than 0.01). However, proglumide did not significantly affect caerulein-stimulated pancreatic protein secretion. Proglumide might be useful in defining the physiological role of CCK.     

细菌Ⅵ型分泌系统的研究进展

Ⅵ型分泌系统(Type Ⅵ secretion system,T6SS)是最近发现的一种新的分泌系统,广泛存在于革兰氏阴性菌变形菌门细菌中,主要由构成分泌系统的结构蛋白、形成跨膜管道结构的转位蛋白、分泌蛋白以及一些对分泌系统起辅助功能的蛋白组成。T6SS能够增强细菌对外界环境的适应性,介导细菌对宿主细胞的致病力以及其他功能。     

细菌的IV型分泌系统

细菌的分泌系统与细菌的生存及致病性密切相关。细菌的分泌系统包括I-VI型,其中,IV型分泌系统是与细菌接合机制有关的一类分泌系统。IV型分泌系统不但可以转运DNA,还可以转运蛋白质及核糖核蛋白复合物等大分子物质,这点区别于其他几种分泌系统。IV型分泌系统介导基因水平转移,通过细菌间接合作用,传递抗性基因和毒力基因,有利于细菌进化;另一方面,IV型分泌系统转运效应蛋白质分子到宿主细胞,参与细菌致病。本文着重从IV型分泌系统几种主要类型的分泌机制等方面对IV型分泌系统进行概述。     

两种吹绵蚧蜡泌物超微结构的研究

采用扫描电镜技术研究了澳洲吹绵蚧lcerya purchase Maskell和埃及吹绵蚧Icerya aegyptiaca(Douglas)不同龄期主要泌蜡腺体和蜡泌物的超微结构.结果发现:成虫期背面的丝状蜡是由多格腺和皮毛共同分泌产生的.卵囊是由雌成虫成熟时腹部腹面的半圆形卵囊带所分泌的,也是实心丝状蜡结构.若虫期吹绵蚧背面覆盖着由多格腺分泌的丝状蜡,实心结构;腹面有长丝状的蜡质缠绕,紧贴腹面体壁的还有一层小蜡圈.     

Selection of CHO host cell subclones with increased specific antibody production rates by repeated cycles of transient transfection and cell sorting

Optimization of host cell lines both for transient and stable protein production is typically hampered by the inherent heterogeneity of cells within a population. This heterogeneity is caused not only by “hard fact” gene mutations, but also by subtle differences in the cellular network of regulation, which may include epigenetic variations. Taking advantage of this heterogeneity, we sorted for naturally occurring variants of CHO‐K1 and CHO‐S host cells that possess an improved cellular machinery for transient antibody production. The long‐term goal of this study was both to identify host cells that yield recombinant cell lines with on average higher productivity, but also to study the molecular differences that characterize such cells, independent of the site of gene integration or gene amplification. To identify such cells we optimized the procedure for transient transfection by electroporation to a degree that gave uniform transfer of plasmid DNA into nearly 100% of the cells and resulted in reproducible average productivities, with a standard deviation of 16% between independent experiments. Using this optimized protocol, the 1% of cells with the highest specific productivity was sorted and subcloned with a cold capture secretion assay. Upon re‐transfection, the resulting subclones showed the same specific productivity as their respective parental cell line. To enrich for cells with potentially stable improved properties, the 1% highest producers were sorted three times, 2 days after transient transfection each, and the enriched population was again sorted into microtiter plates for subcloning. For each of the two parental cell lines tested, three subclones were obtained that had a threefold higher specific productivity after transient transfection. This property was stable for approximately 3 months, indicating that the changes in productivity were regulatory and not mutational. Biotechnol. Bioeng. 2011;108: 386–394. © 2010 Wiley Periodicals, Inc.