悦目金蛛拖牵丝力学性能的变异

蜘蛛丝作为一种具有优良机械性能的天然动物蛋白纤维,其特有的结构和机械性能与其生物学功能密切相关。由大壶状腺纺出的拖牵丝在蜘蛛的行走、建网、捕食、逃生、繁殖等多种生命活动中均发挥了重要的功能,其机械性能会受到多种内外因素相互作用的影响。本文对在不同体重、不同猎物饲养和不同营养状态3种条件下人工抽出的悦目金蛛(Argiope amoena)拖牵丝与其不同单丝间的力学性能进行了比较研究。结果表明,悦目金蛛拖牵丝的力学性能在组间、组内不同个体,以及同一个体不同丝纤维间变异都较大。随着蜘蛛个体的增大,蛛丝横截面直径逐渐增大,这会使得蛛丝的力学性能更好,便于作为救命索的拖牵丝在遇到危险时承受蜘蛛体重;蜘蛛在经过1个月的饥饿后,蛛丝在屈服点附近的力学性能并未发生显著变化,而断裂点应变和断裂能均显著减小,同时也表明无论对于作为救命索还是网丝,拖牵丝的弹性形变性能在与蛛丝相关的微观进化中要优先于塑性形变。这是蜘蛛在能量摄入受到限制时对拖牵丝的投入权衡的结果。     

转蜘蛛拖牵丝蛋白基因家蚕蚕丝氨基酸组成及其机械性能

将以绿色荧光蛋白基因为报告基因、含有人工合成的1.6 kb的蜘蛛拖牵丝蛋白基因及转座子pig-gyBac的转基因载体成功导入减秋无滞育家蚕受精卵,得到转蜘蛛拖牵丝蛋白基因家蚕及绿色荧光茧。对转基因家蚕与对照家蚕丝素蛋白进行了氨基酸组成分析,结果表明转基因蚕茧丝素蛋白甘氨酸和丙氨酸的百分含量分别增加了1.65%和1.80%(平均值);对其生丝的机械性能进行了测试研究,结果表明转基因蚕茧生丝的伸长率降低,断裂强度和初始模量增加,且差异均显著,与理论预期结果吻合。结果表明转基因家蚕蚕丝的机械性能一定程度上得到了提高。     

悦目金蛛拖丝的超微结构研究

采用非固定的抽丝方法,从一只未麻醉的悦目金蛛(Argiope amoena)的纺器将拖丝抽出,然后用扫描电镜(SEM)对拖丝进行超微结构的观察,结果表明:悦目金蛛拖丝至少具有2根、3根、4根及多根单丝纤维构成的4种不同结构,其中有一种类似弹簧的结构;另外,丝的表面还出现一种小环结构,这两种结构可能是拖丝纤维具有优良机械性能的原因之一。"束状结构"和"小环结构"在文献中未见报道。拖丝的直径范围为0.25~10.77μm;悦目金蛛似乎能调节拖丝的结构和直径,以适应其所面临的即时环境。本文基于上述观察结果并结合前人的研究,提出了蜘蛛拖丝结构-生物学功能多样性假说,对蜘蛛丝的结构与生物学功能之间的关系作了探讨。     

兔抗仿蜘蛛牵丝蛋白抗体的制备及应用

将仿蜘蛛牵丝基因s6 0 0克隆到GST融合蛋白表达质粒pGEX KG中 ,利用大肠杆菌表达系统表达并纯化了仿蜘蛛牵丝蛋白S6 0 0 ,以之作为抗原制备了兔抗血清。S6 0 0的氨基酸组分分析与理论值相吻合。蛋白质免疫印迹发现该抗血清能与天然蜘蛛丝反应 ,表明设计的仿蜘蛛丝与天然蜘蛛丝有相似的免疫原性。为了定量检测仿蜘蛛牵丝蛋白在转基因家蚕丝腺中 (或茧壳中 )的表达 ,建立了用ELISA方法定量检测茧壳中仿蜘蛛牵丝蛋白量的工作系统     

棒络新妇和悦目金蛛拖丝超微结构与力学行为

采用SEM对棒络新妇Nephila clavata腹部向上和向下在水平纱窗上爬行时纺出的拖丝、悦目金蛛Argiope amoena捕食拖丝与垂直向下缓慢纺出的拖丝及其圆网的铆钉丝进行了超微结构观察,采用电子单纤强力仪对棒络新妇拖丝与悦目金蛛圆网铆钉丝进行了力学拉伸试验.结果 表明棒络新妇和悦目金蛛拖丝均呈现出一至多根细丝纤维的多样化超微结构特征,其中悦目金蛛圆网铆钉丝还呈现出"S"形似弹簧的结构.两种蜘蛛丝的力学行为和性能与各自的功能要求相一致.蜘蛛能调节拖丝的超微结构、纤维组成和直径大小以适应其在不同环境条件下对力学性能和功能的瞬时需要.研究结果有助于拓宽和加深人们对蜘蛛丝超微结构、力学性能与生物学功能之间关系的认识和理解.     

棒络新妇卵袋结构与组成

采用SEM和氨基酸自动分析仪对棒络新妇 Nephila clavata 卵袋进行了结构观察和氨基酸组成分析.结果 表明棒络新妇的卵袋呈开放的网筛椭球状,结构简单,分为框架(大壶状腺丝)、金黄色或白色外层(主要含大壶状腺丝)和白色内层(主要含柱状腺丝)三部分,内外层都是由大(approx. 6～13 μm)、中(1.25～3.0 μm)、小(approx. 0.7～0.8 μm)直径的丝纤维构成.卵袋框丝和外层丝的氨基酸组成相似,甘氨酸(分别占22.9%和32.5%)和丙氨酸(占24%左右)的含量最丰富,其次是谷氨酸(占16.5%左右)和丝氨酸(分别占8.3%和3%),苏氨酸含量低(分别占2.12%和0.68%).与外层丝相比,卵袋内层丝纤维丝氨酸和苏氨酸的含量显著增加,分别为19.1%和6.11%左右,同时甘氨酸的含量显著减少,为7.2%左右,谷氨酸和脯氨酸的含量较低.大小侧链氨基酸比值(LC:SC)分析表明卵袋内层丝(1.03)﹥卵袋框丝(0.85)＞卵袋外层丝(0.66)≈拖牵丝(0.64)＞蚕丝(0.34),似乎表明卵袋框丝是不同于同纺自大壶状腺的典型蛛丝(拖牵丝)的新型蛛丝纤维.卵袋内层的主要构成成分是另一种具有截然不同氨基酸组成的柱状腺丝.棒络新妇能调节卵袋层丝的颜色、直径、丝纤维的组成及覆盖层的氨基酸组成,使卵袋获得各方面最优化的功能,为卵或若蛛提供支持和保护.     

落叶松(Larix)细根形态特征沿纬度梯度的可塑性

选择5个不同纬度的落叶松人工林,比较了落叶松细根(2 mm)的直径、根长、比根长和比表面积等形态指标、外生菌根及其侵染率的纬度分异特征,分析了形态特征与主要影响因子间的关系。结果表明:随纬度增加,落叶松1、2级根的外生菌根真菌侵染率降低,菌根颜色变深,分叉减少;细根直径变细,根长缩短,细根比根长和比表面积显著增加;同一地点的细根在不同根序以及侵染与未侵染细根间形态差异明显;冗余分析和回归分析显示,细根直径和根长与年均温、年降水、土壤含水量呈显著正相关,而比根长和比表面积则与土壤全碳、土壤养分含量呈正相关。综上,落叶松细根的形态特征沿纬度梯度呈现出有规律的可塑性响应,外生菌根的宏观形态具有明显的地带性差异,直径和根长主要受气候因子影响,而比根长和比表面积则主要与土壤因子有关。     

辽河流域栽培稻对二化螟(Chilo suppressalis)的抗性

采用活体成株鉴定法测定了辽河流域主栽水稻品种对二化螟发育和为害的影响,同时测定了不同品种的主要形态特性、硅细胞数量和主要生化特性,评价这些品种对二化螟的抗性、分析其发生的可能机制。盐丰47对二化螟幼虫表现出较好的抗性,在沈农265上二化螟幼虫体重最小。在所测定的特性中,螟害株率与不同品种倒二叶叶角的自然对数之间具有显著的正相关关系,与稻茎Si含量之间具有显著的负线性相关关系;二化螟幼虫体重只与不同品种稻茎直径之间具有显著的正线性相关关系．研究结果对水稻品种杭件利用和未来水稻抗件育种的意义。     

不同体重对正常饮食下小鼠腹主动脉瘤进展影响的回顾性研究

目的 分析正常饮食情况下,术前体重对猪胰弹性蛋白酶(PPE)诱导的小鼠腹主动脉瘤进展的影响。方法 回顾分析本实验室最近半年44例PPE诱导的小鼠腹主动脉瘤模型中,术前不同体重对后期腹主动脉直径和腹主动脉瘤进展的影响,分析两者的相关性。对术前体重低于25 g的8例入选小鼠(模型组1)和术前体重高于30 g的8例入选小鼠(模型组2)进行体重、脂肪分布、腹主动脉直径、腹主动脉直径变化、腹主动脉瘤病变组织学和免疫组织化学比较。正常阴性对照小鼠未给与任何处理。结果 44例小鼠腹主动脉瘤血管直径和术前体重的相关系数为0.005,未发现体重对腹主动脉瘤进展有显著影响。进一步分析发现,模型1组和模型2组在术后14 d血管直径(1.35 mm vs 1.27 mm)和血管直径变化上(0.85 mm vs 0.72 mm)差异没有显著性。相比正常血管,模型组1和模型组2小鼠在术后14 d,腹主动脉肾下灌注段直径显著增加,全部形成动脉瘤。组织学呈弹性纤维断裂、平滑肌细胞耗竭、炎症反应增加和血管新生异常等典型的腹主动脉瘤病变特征。结论 在正常饮食小鼠,体重未见影响腹主动脉瘤的进展。     

单细胞机械性能检测方法与应用研究进展

细胞机械性能与细胞的生理状态与功能存在密切联系。早期对于细胞机械性能的研究受制于技术条件,只能获得细胞群的弹性或剪切模量,使得少量异质细胞的机械表型被淹没。近年来,单细胞机械性能检测技术得到了蓬勃发展。原子力显微镜、微吸管技术、光镊与光学拉伸、磁扭转流变仪与磁镊等单细胞机械性能检测技术展现出非常高的检测精度,但检测通量相对较低。新型微流控高通量检测方法的出现使检测通量呈几何式增长,有望解决大样本快速检测的需求。本文首先综述原子力显微镜、微吸管、光镊与光学拉伸和磁扭转流变仪与磁镊等单细胞机械性能检测技术。在此基础上,重点介绍细胞过孔、剪切诱导细胞变形和拉伸诱导细胞变形3种新兴微流控高通量检测技术的工作原理及最新研究进展,探讨各类方法的优缺点。最后,本文展望单细胞机械性能检测技术的未来发展方向。     

Consequences of forced silking

The forced silking of a spider to obtain major ampullate (MA) silk for experiments is a standard practice; however, this method may have profound effects on the resulting silk's properties. Experiments were performed to determine the magnitude of the difference in the forces required to draw silk from the MA gland between unrestrained spiders descending on their draglines and restrained spiders from which MA silk was drawn with a motor. The results show that freely falling spiders can spool silk with as little as 0.1 body weights of force, which generates a stress that is about 2% of the silk's tensile strength. In contrast, forcibly silked spiders apply as much as 4 body weights of force with an internal braking mechanism, and this force creates silk stresses in excess of 50% of the silk's tensile strength. The large forces observed in forced silking should strongly affect the draw alignment of the polymer network in the newly spun fibers, and this may account for the differences in material properties observed between naturally spun and forcibly spun MA silks. In addition, the heat produced by the internal friction brake during forced silking may set the upper limit of forced silking speed.     

Biological liquid crystal elastomers

Liquid crystal elastomers (LCEs) have recently been described as a new class of matter. Here we review the evidence for the novel conclusion that the fibrillar collagens and the dragline silks of orb web spiders belong to this remarkable class of materials. Unlike conventional rubbers, LCEs are ordered, rather than disordered, at rest. The identification of these biopolymers as LCEs may have a predictive value. It may explain how collagens and spider dragline silks are assembled. It may provide a detailed explanation for their mechanical properties, accounting for the variation between different members of the collagen family and between the draglines in different spider species. It may provide a basis for the design of biomimetic collagen and dragline silk analogues by genetic engineering, peptide- or classical polymer synthesis. Biological LCEs may exhibit a range of exotic properties already identified in other members of this remarkable class of materials. In this paper, the possibility that other transversely banded fibrillar proteins are also LCEs is discussed.     

Responses of an araneophagic assassin bug,Stenolemus bituberus,to spider draglines

Abstract 1. Animals, as they move through their environment, leave traces of their passage that can be informative to others and convey significant advantages to the animal producing them. However, such traces may also reveal presence, location or identity to enemies. 2. We studied an araneophagic (‘spider‐eating’) assassin bug, Stenolemus bituberus (Heteroptera, Reduviidae), testing whether it associated with areas containing chemotactile traces (e.g. draglines, excreta) left behind by nine sympatric spider species. Stenolemus bituberus were presented with a choice between a substrate containing draglines and a clean substrate. Each hour, for a duration of 12 h, we recorded which substrate was occupied. 3. Stenolemus bituberus tended to associate especially with draglines left by spiders from the genus Achaearanea, their most common prey in nature. 4. These results suggest that S. bituberus exploits draglines from these spiders as cues for indicating prey presence. We also found an increasing tendency to associate with draglines from some spider species through the day, which may be related to circadian patterns or slower response times of some individuals.     

Draglines and Assessment of Fighting Ability in Cannibalistic Jumping Spiders

The frequency of injury and death during female-female aggression varies in the jumping spider genus Portia, with interactions being more violent (likely to end in death or injury of one of the combatants) in P. labiata (from Sri Lanka) than in another two species (P. fimbriata from Australia and P. schultzi from Kenya). To investigate the role of draglines in the assessment of fighting ability, two types of tests were carried out: 1) dragline discrimination and 2) mirror image response (Portia's reaction to mirror images is comparable to interaction with conspecific rivals). For both types of testing, triplets of equal-size conspecific females were used: one female (the test spider) was exposed to draglines of two equal-size conspecific females they had not encountered before (donor spiders). The fighting abilities of donor spiders were determined directly by staging intraspecific contests between them. In dragline-discrimination tests (spider placed in petri dish containing draglines from two conspecific females with different fighting ability), females of P. labiata, but not the other two species, avoided draglines of the superior fighter (i.e., they spent the majority of their time on draglines of donor spiders with lesser fighting ability). For mirror-image testing, the test spider was placed in a petri dish containing a mirror and draglines. Each test spider was tested on two successive days, with donor draglines in the two tests coming from conspecific females with different fighting ability. In these tests, females of P. labiata (but not the other two species) spent less time embracing (each spider pressing its forelegs, palps and front of body against the other spider) and more time in a part of the petri dish where view of the mirror was obstructed when on the draglines of donor spiders with greater fighting ability than when on the other conspecific's draglines. Findings from this study suggest that P. labiata females use signpost cues associated with draglines to assess the relative fighting abilities of unknown opponents.     

雌星豹蛛性信息素的行为学证据

采用星豹蛛(Pardosa astrigera)成熟雄蛛求偶时潜伏时间、静止时间、身体震动和第一墩步足伸展次数等行为参数,利用行为学方法测定了不同性别、日龄和生殖状态的星豹蛛雌蛛释放的拖丝对雄蛛求偶行为的影响。结果表明,雄蛛第一对步足伸展和身体震动等典型求偶行为是进行星豹蛛性信息素生物测定的可靠评价指标。星豹蛛雄蛛能通过拖丝上的性信息素辨别星豹蛛的性别、日龄和生殖状态。雄蛛在成熟3周未交配雌蛛拖丝处理过滤纸上潜伏时间和静止时间都相应最短,在交配未产卵雌蛛、雌亚成蛛和成熟雄蛛拖丝上时间中等,在卵孵化雌蛛拖丝处理滤纸上潜伏时间和静止时间都相应最长。成熟3周未交配雌蛛和交配未产卵雌蛛释放的拖丝都能引起雄蛛第一对步足伸展和身体震动等典型求偶行为,雄蛛对成熟3周未交配雌蛛拖丝典型求偶行为的频率都相应高于交配未产卵雌蛛。卵孵化雌蛛释放的拖丝虽也能引起雄蛛第一对步足伸展行为,但其伸展频率显著降低;而其它拖丝都不能引起雄蛛典型求偶行为。     

Spider capture silk: performance implications of variation in an exceptional biomaterial

Spiders and their silk are an excellent system for connecting the properties of biological materials to organismal ecology. Orb-weaving spiders spin sticky capture threads that are moderately strong but exceptionally extensible, resulting in fibers that can absorb remarkable amounts of energy. These tough fibers are thought to be adapted for arresting flying insects. Using tensile testing, we ask whether patterns can be discerned in the evolution of silk material properties and the ecological uses of spider capture fibers. Here, we present a large comparative data set that allows examination of capture silk properties across orb-weaving spider species. We find that material properties vary greatly across species. Notably, extensibility, strength, and toughness all vary approximately sixfold across species. These material differences, along with variation in fiber size, dictate that the mechanical performance of capture threads, the energy and force required to break fibers, varies by more than an order of magnitude across species. Furthermore, some material and mechanical properties are evolutionarily correlated. For example, species that spin small diameter fibers tend to have tougher silk, suggesting compensation to maintain breaking energy. There is also a negative correlation between strength and extensibility across species, indicating a potential evolutionary trade-off. The different properties of these capture silks should lead to differences in the performance of orb webs during prey capture and help to define feeding niches in spiders.     

Assembly of type I collagen: fusion of fibril subunits and the influence of fibril diameter on mechanical properties.

Structural stability of the extracellular matrix is primarily a consequence of fibrillar collagen and the extent of cross-linking. The relationship between collagen self-assembly, consequent fibrillar shape and mechanical properties remains unclear. Our laboratory developed a model system for the preparation of self-assembled type I collagen fibers with fibrillar substructure mimicking the hierarchical structures of tendon. The present study evaluates the effects of pH and temperature during self-assembly on fibrillar structure, and relates the structural effects of these treatments on the uniaxial tensile mechanical properties of self-assembled collagen fibers. Results of the analysis of fibril diameter distributions and mechanical properties of the fibers formed under the different incubation conditions indicate that fibril diameters grow via the lateral fusion of discrete approximately 4 nm subunits, and that fibril diameter correlates positively with the low strain modulus. Fibril diameter did not correlate with either the ultimate tensile strength or the high strain elastic modulus, which suggests that lateral aggregation and consequently fibril diameter influences mechanical properties during small strain mechanical deformation. We hypothesize that self-assembly is mediated by the formation of fibrillar subunits that laterally and linearly fuse resulting in fibrillar growth. Lateral fusion appears important in generating resistance to deformation at low strain, while linear fusion leading to longer fibrils appears important in the ultimate mechanical properties at high strain.     

Strains in the exoskeleton of spiders

Summary This is a first attempt to measure strain in the exoskeleton of arthropods during locomotion. The selected sites of measurement at the tibia of large spiders allowed a direct estimation of the mechanical input of slit sense organs, which are biological strain receptors. This study includes (I) the development of techniques for measurement, (II) an anatomical and biomechanical investigation of the tibia-metatarsus joint of spiders, (III) the measurement of cuticular strains developed by external and internal forces in tethered theraphosids (Aviculariidae), and (IV) the measurement of strain in the legs of freely moving animals.