A simple assay to study social behavior in Drosophila: measurement of social space within a group

We have established a new simple behavioral paradigm in Drosophila melanogaster to determine how genes and the environment influence the behavior of flies within a social group. Specifically, we measure social space as the distance between two flies. The majority of Canton-s flies, regardless of their gender, are within two body lengths from each other. Their social experience affects this behavior, with social isolation reducing and mating enhancing social space respectively, in both males and females. Unlike several other social behaviors in the fly, including the formation of social groups themselves (a well-described behavior), social space does not require the perception of the previously identified aggregation pheromone cis-vaccenyl acetate. Conversely, performance of the assay in darkness or mutations in the eye pigmentation gene white increased social space. Our results establish a new assay for the genetic dissection of a fundamental mode of social interaction.     

鹭兰种子无菌繁殖球根

为建立鹭兰（Habenaria radiata）种子繁殖法,对无菌培养条件下的种子发芽和幼苗生长进行了观测,并对无菌繁殖获得的球根进行了种植试验。结果表明,鹭兰种子不进行处理难以无菌培养发芽,种子通过75%乙醇10 s+1% NaClO 10 min处理,或者40 kHz超声波2.5 min+1% NaClO 10 min处理,在1/2MS培养基的发芽率可达58%以上;在1/2MS培养基中,添加GA-3 1 mg·L^-1对叶片生长有促进作用,添加IAA 1 mg·L^-1对根生长和球根形成有促进作用,球根形成率达55%;无菌繁殖的球根贮存至春季盆栽,成苗率约72%。     

Cruising specialists and accelerators--are different types of fish locomotion driven by differently structured myosepta?

Locomotor specialists, such as accelerators and cruisers, have clearly differing body designs. For physical reasons these designs are mutually exclusive, i.e. cruisers necessarily have poor accelerating capabilities and vice versa. For the first time, we examine whether differences in the anatomy of the musculo-tendinous system of the trunk are present in addition to the differences in external body design. We investigated the myoseptal series of two closely related locomotor specialists, the cruiser Scomber scombrus and the accelerator Channa obscura, by microdissections combined with polarized light microscopy and histology. Our comparison includes 3D-morphology of myosepta, spatial arrangement and length of myoseptal tendons, their relation to red and white muscles, rostrocaudal changes in all these aspects and the musculo-tendinous system of the caudal fin. Regarding all these features, Channa has retained the plesiomorphic condition of its actinopterygian ancestor. In contrast, the derived morphology of Scomber is characterized by (i) lateral (LT) and myorhabdoid tendons (MT) that are lengthened to up to 20% of body length (compared to a maximum of 8.2% in Channa), (ii) posterior myoseptal cones that are subsequently linked by horizontal projections of merged LTs and MTs, (iii) an increased area of red muscle fibers that insert to LTs of myosepta, (iv) the reduction of epineural (ENTs) and epipleural tendons (EPTs) that connect backbone and skin, (v) specific caudal tendons that are identified to be serial homologues of LTs and MTs of more anterior myosepta, (vi) and a partial reduction of intrinsic caudal muscles. These results suggest the following functional adaptations in the cruiser Scomber. Red muscle forces may be transmitted through LTs and posterior cones to the prominent tendons of the caudal fin. The length of LTs and the intersegmental connections along the posterior cones may facilitate posterior force transmission and may be correlated with the long propulsive wavelength generally observed in cruising carangiform swimmers. Epineural and epipleural tendons are interpreted to minimize lateral backbone displacement during high body curvatures. This is consistent with the lack of these tendons in Scomber, because high body curvatures are not displayed in stiffer-bodied carangiform swimmers. It remains to be tested whether the specializations revealed in this initial study for Scomber represent general specializations of carangiform swimmers. Taking into account the geometry of myoseptal tendons and the horizontal septum we evaluate how local bending according to beam-theory can be generated by white or red muscle activity in Channa and Scomber. In both species, the musculo-tendinous anatomy of the caudal fin explains the functional asymmetry of the caudal fin that was experimentally revealed in previous studies.     

CRISPR相关生物信息学基础

成簇规律间隔短回文序列（clustered regularly interspaced short palindromic repeats,CRISPR）是细菌和古细菌在不断进化的过程中获得的一种适应性免疫防御机制,该结构与一些功能相关的蛋白质（CRISPR associated, Cas）合称CRISPR Cas系统。由于其致突变效率高、操作简单及成本较低的特点,近年来对CRISPR/Cas系统的研究获得越来越广泛的关注。该系统迅速在各领域中得到广泛应用,被认为是一种具有广阔应用前景的基因组定点改造分子工具。但是,该系统存在脱靶效应、测序数据分析等挑战。为此,许多研究者开发出各种软件解决以上问题。本文着重从生物信息学的角度出发,对CRISPR/Cas系统中sgRNA的设计软件、CRISPR全基因组筛选功能基因的测序数据分析软件以及CRISPR在生物信息学中的运用作一系统综述。     

EDAR参与绵羊毛发性状与生长的调节

绵羊的背部、耳部和腹股沟部的毛发性状、生长速度存在差异,背部毛发弯曲、细长、密度高、生长速度快,耳部、腹股沟部毛发粗直、密度低、生长速度慢。研究表明,EDA/EDAR、IGFBP5/Krox 20、WNT等介导的信号通路及毛发角蛋白基因对毛发纤维弯曲的形成有重要的影响。本研究利用实时荧光定量PCR、原位杂交技术、Western 印迹和免疫组织化学等技术,对外异蛋白受体（ectodysplasin A receptor,EDAR）在绵羊背部、耳部和腹股沟皮肤中的mRNA、蛋白质表达水平和定位进行研究,以探讨EDAR与绵羊毛发的生长和性状的关系。实时荧光定量PCR结果显示,EDAR在绵羊背部皮肤中相对基因表达量是绵羊腹股沟皮肤的4.9倍(P＜ 0.01),耳部是腹股沟部的1.4倍（P＜0.05）,背部是耳部的3.4倍（P＜0.05）;原位杂交和免疫组化结果表明,EDAR基因mRNA和蛋白质在背部、耳部和腹股沟部毛囊均有表达。根据光密度值可知,背部表达量最高,耳部次之,而腹股沟部最低;Western 印迹结果显示,绵羊皮肤组织蛋白质提取物中存在与兔抗EDAR多克隆抗体发生免疫阳性反应的蛋白条带,绵羊皮肤背部平均蛋白质表达量最高,耳部次之,而腹股沟部最低,差异极显著(P ＜ 0.01)。研究结果提示,EDAR可能参与绵羊毛发卷曲的形成和调控,对毛发密度、生长速度等可能也有影响。     

Conserved folding pathways of alpha-lactalbumin and lysozyme revealed by kinetic CD, fluorescence, NMR, and interrupted refolding experiments

In this report, it is shown by a combination of stopped-flow CD, fluorescence, and time-resolved NMR studies that the Ca^2 +-induced refolding of bovine α-lactalbumin (BLA) at constant denaturant concentration (4 M urea) exhibits triple-exponential kinetics. In order to distinguish between parallel folding pathways and a strictly sequential formation of the native state, interrupted refolding experiments were conducted. We show here that the Ca^2 +-induced refolding of BLA involves parallel pathways and the transient formation of a folding intermediate on the millisecond timescale. Our data furthermore suggest that the two structurally homologous proteins BLA and hen egg white lysozyme share a common folding mechanism. We provide evidence that the guiding role of long-range interactions in the unfolded state of lysozyme in mediating intersubdomain interactions during folding is replaced in the case of BLA by the Ca^2 +-binding site. Time-resolved NMR spectroscopy, in combination with fast ion release from caged compounds, enables the measurement of complex protein folding kinetics at protein concentrations as low as 100 μM and the concomitant detection of conformational transitions with rate constants of up to 8 s^− 1.     

Does Eulemur cinereiceps exist? Preliminary evidence from genetics and ground surveys in southeastern Madagascar

Although appearing in the literature as early as 1890, the brown lemur form Eulemur cinereiceps has recently resurfaced as a potentially valid taxon, distinct from neighboring, presumably closely related species such as white-collared lemurs (Eulemur albocollaris). We propose two scenarios for the potential separation of E. cinereiceps and E. albocollaris: (1) coastal and interior populations represent two distinct taxa and (2) the coastal population north of the Manampatrana River (the locality for purported museum specimens of E. cinereiceps) represents a distinct species from E. albocollaris found south of the river and in the interior escarpment forests. We tested these hypotheses using data from ground surveys and genetic sampling. Surveys were conducted in coastal forest fragments both north and south of the Manampatrana River in July-August 2006. Genetic samples were collected at two coastal sites and one interior forest. We used maximum parsimony, maximum likelihood, and neighbor-joining analyses on mitochondrial DNA regions to determine if populations from different sites clustered into diagnosable clades. Results from field surveys confirmed the presence of forms commonly referred to as E. albocollaris at the two southern coastal forests; no consistent phenotypic differences across sites were observed. All genetic analyses yielded identical results: coastal and interior populations do not cluster into separate groups, thus rejecting the first hypothesis. Eulemur species and all other day-active lemurs have apparently been extirpated from coastal forests north of the Manampatrana. Owing to the absence of lemurs from the northern coastal localities, we could not conclusively support or reject the second scenario. However, based on examination of the original plates and museum specimens, as well as the biogeographic patterns typical of this region, we strongly suspect that all populations from this area belong to a single species. We conclude with remarks regarding the apparent priority of E. cinereiceps for this taxon.     

活化的T细胞核内因子（NFAT）的调节及其在神经系统中的功能

活化的T细胞核内因子（nuclear factor of activated T-cells, NFAT）作为细胞信号转导通路中的一类重要的转录因子参与细胞功能的调节. NFAT的活化主要是通过细胞内钙/钙调神经磷酸酶（Ca2+/calcineurin）的刺激启动,它脱磷酸后发生核转位并与DNA的特定序列结合,同时通过与其它转录因子的协同作用,调节目的基因的特定表达. NFAT在免疫系统中所调节的基因表达已经得到了充分的研究. 近年实验研究发现,NFAT的转录因子家族在脊椎动物的神经系统中也发挥着非常重要的作用. 本文综述了NFAT家族蛋白的分类、结构、磷酸酶与激酶对其出入核的调节及在神经系统中的研究进展,使得能够更加全面地认识calcineurin/NFAT信号通路的作用. 此外,由于环孢菌素A（cyclosporin A）等药物在神经系统应用的局限性,对于NFAT调节深入研究,也将为筛选或者开发更为高效、低毒药物提供新的思路.     

Soil Phosphorus Uptake by Continuously Cropped Lupinus albus: A New Microcosm Design

When grown in soils with sparingly available phosphorus (P), white lupin (Lupinus albus L.) forms special root structures, called cluster roots, which secrete large amounts of organic acids and concomitantly acidify the rhizosphere. Many studies dealing with the understanding of this P acquisition strategy have been performed in short time experiments either in hydroponic cultures or in small microcosm designs with sand or sand:soil mixtures. In the present study, we applied an experimental design which came nearer to the natural field conditions: we performed a one-year experiment on large microcosms containing 7 kg of soil and allowing separation of rhizosphere soil and bulk soil. We planted six successive generations of lupins and analysed P uptake, organic P desorption, phosphatase activities and organic acid concentrations in different soil samples along a spatio-temporal gradient. We compared the rhizosphere soil samples of cluster (RSC) and non-cluster roots (RSNC) as well as the bulk soil (BS) samples. A total shoot biomass of 55.69 ± 1.51 g (d.w.) y⁻¹ was produced and P uptake reached 220.59 ± 5.99 mg y⁻¹. More P was desorbed from RSC than from RSNC or BS (P < 0.05). RSC and RSNC showed a higher activity of acid and alkaline phosphatases than BS samples and a higher acid phosphatase activity was observed in RSC than in RSNC throughout the one-year experiment. Fumarate was the most abundant organic acid in all rhizosphere soil samples. Citrate was only present in detectable amounts in RSC while malate and fumarate were recovered from both RSC and RSNC. Almost no organic acids could be detected in the BS samples. Our results demonstrated that over a one-year cultivation period in the absence of an external P supply, white lupin was able to acquire phosphate from the soil and that the processes leading to this P uptake took place preferentially in the rhizosphere of cluster roots.