Knock-down of LAR protein tyrosine phosphatase induces insulin resistance

To test the role of the leukocyte common antigen-related protein tyrosine phosphatase (LAR) as a regulator of insulin receptor (IR) signalling, an siRNA probe against LAR was developed. Knock-down of LAR induced post-receptor insulin resistance with the insulin-induced activation of PKB/Akt and MAP kinases markedly inhibited. The phosphorylation and dephosphorylation of the IR and insulin receptor substrate (IRS) proteins were unaffected by LAR knock-down. These results identify LAR as a crucial regulator of the sensitivity of two key insulin signalling pathways to insulin. Moreover, the siRNA probe provides a molecular tool of general applicability for further dissecting the precise targets and roles of LAR.     

大黄葸醌提取物对建鲤抗应激及生长的影响

将750尾建鲤随机分成5组,第1组为对照组,投喂基础日粮,另外4组为试验组,投喂基础日粮中分别添加0．5％、1．0％、2．0％、4．0％大黄蒽醌提取物。从2005年7月到9月连续投喂70d后,对鱼体进行高密度应激,测定其生长、应激前后血液皮质醇、血糖、溶菌酶等变化。结果表明：与对照组相比,应激前添加大黄葸醌提取物提高了鱼体增重率、特定生长率、鱼体丰满度、血液溶菌酶活性,降低了饵料系数与鱼体死亡率,但是与大黄葸醌提取物的添加水平不成线性关系,其中2．0％试验组还显著降低了血液皮质醇。高密度应激1d后,各组血液中的皮质醇、血糖、溶菌酶都有不同程度的增加,其中对照组最高,添加1．0％和2．0％大黄蒽醌提取物的试验组相对较低。应激前后各组鱼的攻毒试验表明：除应激前添加1．0％和2．0％大黄蒽醌提取物的组没有死鱼外,其它各组都有死鱼,其中对照组死亡率最高。因此添加1．0％-2．0％大黄蒽醌提取物提高了机体抗应激能力,并对病原菌感染起一定的保护作用,促进了鱼体生长。     

Phylogeography and alpha taxonomy of the common dolphin (Delphinus sp.)

The resolution of taxonomic classifications for delphinid cetaceans has been problematic, especially for species in the genera Delphinus, Tursiops and Stenella. The frequent lack of correspondence between morphological and genetic differentiation in these species raises questions about the mechanisms responsible for their evolution. In this study we focus on the genus Delphinus, and use molecular markers to address questions about speciation and the evolution of population structure. Delphinus species have a worldwide distribution and show a high degree of morphological variation. Two distinct morphotypes, long-beaked and short-beaked, have been considered different species named D. capensis and D. delphis, respectively. However, genetic differentiation between these two forms has only been demonstrated in the Pacific. We analysed samples from eight different geographical regions, including two morphologically defined long-beaked form populations, and compared these with the eastern North Pacific populations. We found high differentiation among the populations described as long-beaked instead of the expected monophyly, suggesting that these populations may have evolved from independent events converging on the same morphotype. We observed low genetic differentiation among the short-beaked populations across a large geographical scale. We interpret these phylogeographical patterns in the context of life history and population structure in related species.     

Growth and reproduction in the Icelandic common seal (Phoca vitulina L., 1758)

Length, weight and maturity were studied in relation to age in the common seal (Phoca vitulina vitulina L., 1758), collected during the periods 1979-1983 and 1990-2000 in Icelandic waters. The maximum age of common seal observed was 36 years for females and 30 years for males. For common seal females and males, asymptotic length and weight were 161 cm and 93 kg and 174 cm and 97 kg, respectively, showing slight sexual dimorphism in size. The condition of adult females, measured as fat thickness at the lower end of the sternum, was lower in the period 1979-1983 than in 1990-2000 during June-September, the breeding and mating time of the Icelandic common seal. Males reached sexual maturity between 5 and 7 years, whereas 50% of females did so at age 4 years. Including the length and age interaction term in the logistic regression model for the maturity of females significantly improved it. Thus, body size matters in the onset of maturity. The mean birthing date for the Icelandic common seal was found to be in early June. A comparison of animals collected in the two periods 1979-1983 and 1990-2000 did not show significant differences in growth and the average age of sexual maturity for either males or females. The observed decline of the Icelandic common seal population is most probably caused by increased mortality, due to exploitation and accidental by-catch in gill-nets, rather than a decrease in fecundity.     

Intraspecific conflict over host manipulation between different larval stages of an acanthocephalan parasite

Competitive interactions between coinfecting parasites are expected to be strong when they affect transmission success. When transmission is enhanced by altering host behaviour, intraspecific conflict can lead to 'coinfection exclusion' by the first-in parasite or to a 'sabotage' of behavioural manipulation by the youngest noninfective parasite. We tested these hypotheses in the acanthocephalan parasite Pomphorhynchus laevis, reversing phototaxis in its intermediate host Gammarus pulex. No evidence was found for coinfection exclusion in gammarids sequentially exposed to infection. Behavioural manipulation was slightly weakened but not cancelled in gammarids infected with mixed larval stages. Therefore, coinfecting infective and noninfective larvae both suffered competition, potentially resulting in delayed transmission and increased risk of mortality, respectively. Consequently, noninfective larva is not just a 'passive passenger' in the manipulated host, which raises interesting questions about the selective pressures at play and the mechanisms underlying manipulation.     

Liprin-α1 affects the distribution of low-affinity β1 integrins and stabilizes their permanence at the cell surface

Integrins mediate the interaction between cells and extracellular matrix by assembling adhesive structures that need to be dynamically modulated to allow cell motility. We have recently identified liprin-α1 as an essential regulator of integrin dynamics required for efficient cell motility. Here we investigated the effects of liprin-α1 expression on β1 integrin receptors. We found that increased levels of liprin-α1 affected the localization of inactive, low-affinity integrins, while increasing the average size of β1 integrin-positive focal adhesions. Although a direct interaction between β1 integrins and liprin-α1 could not be revealed biochemically, a striking colocalization between redistributed inactive β1 integrins and liprin-α1 was observed. The tight association of overexpressed and endogenous liprin-α1 to the cytoplasmic side of the ventral plasma membrane suggested a possible role of liprin in stabilizing integrin receptors at the cell surface. In support of this hypothesis, we demonstrated an inhibitory effect of liprin overexpression on antibody-induced β1 integrin internalization. On the other hand, depletion of endogenous liprin-α by small interfering RNA increased the rate of integrin internalization. Overall, these results support the hypothesis that liprin-α1 exerts its action on focal adhesion turnover by influencing the localization and stability of integrin receptors at the cell surface.     

Mitochondrial respiration defects modulate differentiation but not proliferation of hematopoietic stem and progenitor cells

Mitochondrial energy production is involved in various cellular processes. Here we show that ATP content is significantly increased in lineage-restricted progenitor cells compared with hematopoietic stem and progenitor cells (HSPCs) or more differentiated cells. Transplantation analysis using a mouse model of mitochondrial disease revealed that mitochondrial respiration defects resulted in a significant decrease in the total number and repopulating activity of bone marrow cells, although the number of HSPCs increased. The proliferative activity of HSPCs and lineage-restricted progenitor cells was not impaired by reduction of ATP content and there seems to be no associated increase in reactive oxygen species levels and apoptosis. Our findings indicate that mitochondrial respiration defects modulate HSPC commitment/differentiation into lineage-restricted progenitor cells.     

Elucidation of the speciation history of three sister species of crown-of-thorns starfish (Acanthaster spp.) based on genomic analysis

The crown-of-thorns starfish (COTS) is a coral predator that is widely distributed in Indo-Pacific Oceans. A previous phylogenetic study using partial mitochondrial sequences suggested that COTS had diverged into four distinct species, but a nuclear genome-based analysis to confirm this was not conducted. To address this, COTS species nuclear genome sequences were analysed here, sequencing Northern Indian Ocean (NIO) and Red Sea (RS) species genomes for the first time, followed by a comparative analysis with the Pacific Ocean (PO) species. Phylogenetic analysis and ADMIXTURE analysis revealed clear divergences between the three COTS species. Furthermore, within the PO species, the phylogenetic position of the Hawaiian sample was further away from the other Pacific-derived samples than expected based on the mitochondrial data, suggesting that it may be a PO subspecies. The pairwise sequentially Markovian coalescent model showed that the trajectories of the population size diverged by region during the Mid-Pleistocene transition when the sea-level was dramatically decreased, strongly suggesting that the three COTS species experienced allopatric speciation. Analysis of the orthologues indicated that there were remarkable genes with species-specific positive selection in the genomes of the PO and RS species, which suggested that there may be local adaptations in the COTS species.     

A phylogeographic study of the cosmopolitan genus Phragmites (Poaceae) based on AFLPs

Within the genus Phragmites (Poaceae), the species P. australis (the common reed) is virtually cosmopolitan, and shows considerable variation in ploidy level and morphology. Genetic variation in Phragmites was studied using AFLPs, and analysed with parsimony and distance methods. Groups of P. australis strongly supported in the analyses include one that comprises all South American clones, a distinct group from the US Gulf Coast, and a group of E. Asian and Australian octoploids. Among the other species, the paleotropical P. vallatoria is supported as monophyletic and most closely related to the paraphyletic P. mauritianus and to the Gulf Coast and S. American groups. The E. Asian species P. japonicus is closely related to a group of P. australis clones mostly from central North America. Tetraploidy predominates in the genus, and optimisation of chromosome numbers onto the phylogeny shows that higher ploidy levels have evolved many times.     

Old fold in a new X‐ray diffraction dataset? Low‐resolution molecular replacement using representative structural templates can provide phase information

The advent of structural genomics has led to a dramatic increase in the number of structures deposited in the Protein Data Bank. The number of new folds, however, still remains a very small fraction of the total number of deposited structures. Recent data on the progress of the structural genomics initiative reveals that more than 85% of target proteins that progress to the stage of data collection and structure determination have a known fold. Enzymes, which tend to exploit reaction space while adopting a common stable scaffold, contribute significantly to this observation. Herein, we evaluate a method to examine the "old fold in a new dataset" scenario likely to be encountered in the structural genomics pipeline. We demonstrate that a fold detection strategy based on secondary structure signatures followed by molecular replacement using a minimalist model can be effectively used to solve the phase problem in X-ray crystallography without further recourse to heavy atom derivatives or multiple anomalous dispersion techniques. Three common folds-the triosephosphate isomerase (TIM), adenine nucleotide alpha hydrolase-like (HUP), and RNA recognition motif (RRM)-were examined using this approach. The results presented herein also provide an estimate of the extent of phase information that can be derived from a single domain in a large multidomain structure.