棒络新妇和悦目金蛛丝腺形态初步观察

研究比较了结网型蜘蛛棒络新妇Nephila clavata和悦目金蛛Argiope amoena的丝腺形态特征,为国内蜘蛛丝腺蛋白的研究提供原始的丝腺解剖图,同时结合对2种蜘蛛卵袋的解剖、网的特征和室内捕食黄粉虫Tenebrio molitor幼虫行为的观察比较,探讨了2种蜘蛛丝腺的生物学功能与其生存繁殖策略之间的关系。本文分别观察描述了棒络新妇和悦目金蛛的大壶状腺、小壶状腺、鞭状腺、柱状腺、葡萄状腺和梨状腺共6种丝腺。2种蜘蛛丝腺形态特征基本相似;部分丝腺在形态结构和颜色上有些差异;悦目金蛛的葡萄状腺比棒络新妇发达。观察表明2种蜘蛛的网和卵袋特征差异较大,两者捕食策略也不同,棒络新妇采用咬一捆缚（Bit—Wrapping）策略,悦目金蛛则采用捆缚一咬（Wrapping-Bit）策略。棒络新妇和悦目金蛛的网和卵袋特征与丝腺的颜色相一致。同时,其葡萄状腺数量和大小与其各自的捕食策略相关。     

三种类型蜘蛛丝的结构及生物学功能

利用付里叶变换红外光谱仪(FFIR)对棒络新妇(Nephila clavata)、悦目金蛛(Argiope amoena)的大壶状腺丝(拖丝)、悦目金蛛的捕丝(粘性螺旋丝)和卵袋丝这3种不同类型蜘蛛丝的二级结构进行了测试研究。结果表明：蜘蛛丝同时包含无规则卷曲、α-螺旋和β-折叠构象;对这3种蛛丝的红外光谱进行比较表明同一蜘蛛的不同类型蛛丝所含的这3种二级结构的比例不同,这种不同组成的二级结构就赋予了蜘蛛丝不同的特性,这种特性又与其不同的功能相适应。此外,还用扫描电镜(SEM)和光学显微镜对悦目金蛛和小悦目金蛛(A．minuta)的拖丝和捕丝做了形态结构观察。蜘蛛丝这种天然动物蛋白纤维所具有的特殊的形态结构、蛋白质二级结构与其特殊的性能和生物学功能是高度一致的。     

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。     

悦目金蛛3种蛛丝超微结构和理化特性的初步研究

利用扫描电镜、氨基酸分析仪、X-衍射仪和单纤维电子强力仪分别对悦目金蛛Argiopeamoena拖丝、网框丝和卵袋丝的超微结构和理化特性进行了测试和观察。结果表明,悦目金蛛卵袋不是由一种结构均一的丝纤维构成,而是由直径相差悬殊的Ⅰ型卵袋丝和Ⅱ型卵袋丝2种丝纤维共同组成,该结果对卵袋丝仅由管状腺产生的观点提出了疑问。在氨基酸组成上悦目金蛛拖丝和网框丝相似,但其卵袋丝的氨基酸组成与拖丝和网框丝相比差别明显。另外还发现卵袋丝的强度、结晶度大于拖丝和网框丝,而它的延伸性能却不及拖丝和网框丝。     

悦目金蛛和棒络新妇卵袋丝物理化学结构表征及其力学性能研究

蜘蛛丝是一种具有优良机械性能的天然动物蛋白纤维,它特有的结构和性能与其生物学功能密切相关。作者采用氨基酸自动分析仪、傅立叶转换红外光谱仪、扫描电镜和电子单纤强力仪对悦目金蛛（Argiope amoena）和棒络新妇（Nephila clavata）的卵袋丝进行了物理化学结构表征与力学性能的研究,结果表明两种蜘蛛卵袋均由微米级柱状腺丝、大壶状腺丝、亚微米级或纳米级葡萄状腺丝构成。卵袋丝的表面形貌特征、极性氨基酸含量、大侧链与小侧链氨基酸的比值、无定型区、β-折叠结构与结晶结构的含量等氨基酸组成种类与蛋白质二级结构特征,均满足各自生物学功能对断裂强度、延展性、初始模量等力学性能的要求。     

大腹园蛛拖丝蛋白一级结构初步研究

采用部分酸水解的方法对大腹园蛛（Ａｒａｎｅｕｓｖｅｎｔｒｏｃｏｓｕｓ）拖丝纤维蛋白进行水解,通过反相高液相色谱分离到一些重复小肽片段,对其序列分析表明,该蜘蛛拖丝蛋白与肖蛸科的棒络新妇蛛（Ｎｅｐｈｉｌａｃｌａｖｉｐｅｓ）丝蛋白的序列比较,有一个完全相同的肽段ＧＹＧＰＧ,其余所测片段测存在差异,显示不同种属的蜘蛛丝在一级结构上的同异,并探讨了结构和功能的关系。     

4种常见蜘蛛蛛丝光学显微镜观察及机械性能的测定

对4种常见蜘蛛大腹园蛛Araneus ventricosus、迷宫漏斗蛛Agelena labyrinthica、草间钻头蛛Hylyphantes graminicola和棒络新妇Nephila clavata蛛丝的物理特性和机械性能进行了初步研究.结果 表明:捕丝是由2根核心丝构成,4种蛛丝呈现不同的颜色;4种蜘蛛蛛丝的密度略有不同,棒络新妇的蛛丝密度最大,草间钻头蛛的蛛丝密度最小;蛛丝中棒络新妇的断裂伸长率和断裂强度都最大,分别为47.3%和852.6 Nmm-2,草间钻头蛛蛛丝的断裂伸长率和断裂强度都最小,分别为32.1%和652.3 Nmm-2.与其它物理化学材料相比,蛛丝具有优异的综合力学性能,生物学特性与生物学功能具有高度一致性.     

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。     

横纹金蛛捕食经验对其丝纤维力学行为与性能的影响

蛛网是蜘蛛的捕食工具,蛛网网丝的结构与性能不仅影响蜘蛛的捕食效率,也关系着蜘蛛的捕食投入。本文利用单纤维电子强力仪研究横纹金蛛(Argiope bruennichi)室内捕食面包虫(Tenebrio molitor)时上前与返回捕食拖丝的力学性能以及捕食经验对圆网半径丝修补前后的力学性能的影响。结果表明,与上前捕食拖丝相比,返回捕食拖丝减小了弹性区的投入,增加了屈服区和加强区的投入,且返回时捕食拖丝更具柔韧性。整体而言,与初始半径丝相比,在未喂食面包虫的条件下,网的半径修补丝增加了力学性能的投入;而在喂食面包虫的条件下,网的半径修补丝减少了力学性能的投入。所测试丝样中出现了两种类型的蛛丝力学行为:一种为典型的蛛丝力学行为;另外一种为似黏流性材料的力学行为,其反映的是满足蛛丝耗散猎物或自身下降时动能的另外一种力学性能的策略。本研究表明蜘蛛能根据其捕食经验遵循Cost-Benefit原则对蜘蛛丝的力学性能进行调节,从而调整捕食投入。     

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.     

2种织圆网蜘蛛个体大小与蛛网粘丝区面积间关系

在不惊扰蜘蛛的情况下对悦目金蛛Argiope amoena和圆尾肖蛸Tetragnatha vermiformis的个体大小与蛛网粘丝区面积问的相关性进行了野外观察。主要测量了蜘蛛体长、蛛网粘丝区直径、蛛网框丝固着点间最大距离、网中枢到地面或水面距离、网平面与水平线的夹角等参数;另外还对蛛网框丝固着点间最大距离作为圆网蛛网址选择行为量化指标的可行性进行了探讨。结果表明,2种蜘蛛体长与蛛网粘丝区面积以及蜘蛛体长与蛛网框丝固着点间最大距离均呈现明显的正相关。悦目金蛛体长与蛛网粘丝区面积的相关性高于圜尾肖蛸,这可能与悦目金蛛在其生境中易于找到框丝固着点、而圆尾肖蛸在其生境中较难找到框丝同着点有关;     

Polarized light microscopy, variability in spider silk diameters, and the mechanical characterization of spider silk

Abstract. Spider silks possess a remarkable combination of high tensile strength and extensibility that makes them among the toughest materials known. Despite the potential exploitation of these properties in biotechnology, very few silks have ever been characterized mechanically. This is due in part to the difficulty of measuring the thin diameters of silk fibers. The largest silk fibers are only 5–10 μm in diameter and some can be as fine as 50 nm in diameter. Such narrow diameters, coupled with the refraction of light due to the anisotropic nature of crystalline regions within silk fibers, make it difficult to determine the size of silk fibers. Here, we report upon a technique that uses polarized light microscopy (PLM) to accurately and precisely characterize the diameters of spider silk fibers. We found that polarized light microscopy is as precise as scanning electron microscopy (SEM) across repeated measurements of individual samples of silk and resulted in mean diameters that were ～0.10 μm larger than those from SEM. Furthermore, we demonstrate that thread diameters within webs of individual spiders can vary by as much as 600%. Therefore, the ability of PLM to non‐invasively characterize the diameters of each individual silk fiber used in mechanical tests can provide a crucial control for natural variation in silk diameters, both within webs and among spiders.     

Spinning an elastic ribbon of spider silk

The Sicarid spider Loxosceles laeta spins broad but very thin ribbons of elastic silk that it uses to form a retreat and to capture prey. A structural investigation into this spider's silk and spinning apparatus shows that these ribbons are spun from a gland homologous to the major ampullate gland of orb web spiders. The Loxosceles gland is constructed from the same basic parts (separate transverse zones in the gland, a duct and spigot) as other spider silk glands but construction details are highly specialized. These differences are thought to relate to different ways of spinning silk in the two groups of spiders. Loxosceles uses conventional die extrusion, feeding a liquid dope (spinning solution) to the slit-like die to form a flat ribbon, while orb web spiders use an extrusion process in which the silk dope is processed in an elongated duct to produce a cylindrical thread. This is achieved by the combination of an initial internal draw down, well inside the duct, and a final draw down, after the silk has left the spigot. The spinning mechanism in Loxosceles may be more ancestral.     

Does the Giant Wood Spider Nephila pilipes Respond to Prey Variation by Altering Web or Silk Properties?

Recent studies demonstrated that orb‐weaving spiders may alter web architectures, the amount of silk in webs, or the protein composition of silks in response to variation in amount or type of prey. In this study, we conducted food manipulations to examine three mechanisms by which orb‐weaving spiders may adjust the performance of webs to variation in prey by altering the architectures of webs, making structural changes to the diameters of silk threads, and manipulating the material properties or amino acid composition of silk fibers. We fed Nephila pilipes two different types of prey, crickets or flies, and then compared orb structure and the chemical and physical properties of major ampullate (MA) silk between groups. Prey type did not affect orb structures in N. pilipes, except for mesh size. However, MA silk diameter and the stiffness of orbs constructed by spiders fed crickets were significantly greater than for the fly group. MA fibers forcibly silked from N. pilipes fed crickets was significantly thicker, but less stiff, than silk from spiders fed flies. Spiders in the cricket treatment also produced MA silk with slightly, but statistically significantly, more serine than silk from spiders in the fly treatment. Percentages of other major amino acids (proline, glycine, and glutamine) did not differ between treatments. This study demonstrated that orb‐weaving spiders can simultaneously alter some structural and material properties of MA silk, as well as the physical characteristics of webs, in response to different types of prey.     

Short and long range order of the morphology of silk from Latrodectus hesperus (Black Widow) as characterized by atomic force microscopy.

The surfaces of both stretched and unstretched silk threads from the cobweb weaver, Latrodectus hesperus (Black Widow) have been examined by atomic force microscopy (AFM). AFM images of cobweb scaffolding threads show both unordered and highly ordered regions. Two types of fibers within the threads were observed: thicker (approximately 300 nm in diameter) fibers oriented parallel to the thread axis and thinner (10-100 nm) fibrils oriented across the thread axis. While regions which lacked parallel fibers or fibrils were observed on threads at all strain values, the probability of observing fibers and/or fibrils increased with strain. High-resolution AFM images show that with increasing strain, both mean fiber and fibril diameters decrease and that fibrils align themselves more closely with the thread axis. The observation of fibers and fibrils within the cobweb threads has implications for current models of the secondary and tertiary structure and organization of spider silk.     

Novel assembly properties of recombinant spider dragline silk proteins

Spider dragline silk, which exhibits extraordinary strength and toughness, is primarily composed of two related proteins that largely consist of repetitive sequences. In most spiders, the repetitive region of one of these proteins is rich in prolines, which are not present in the repetitive region of the other. The absence of prolines in one component was previously speculated to be essential for the thread structure. Here, we analyzed dragline proteins of the garden spider Araneus diadematus, ADF-3 and ADF-4, which are both proline rich, by employing the baculovirus expression system. Whereas ADF-3 represented an intrinsically soluble protein, ADF-4 was insoluble in vitro and self-assembled into filaments in the cytosol of the host insect cells. These ADF-4 filaments displayed the exceptional chemical stability of authentic silk threads. We provide evidence that the observed properties of ADF-3 and ADF-4 strongly depend on intrinsic characteristics such as hydropathicity, which differs dramatically between the two proteins, as in most other pairs of dragline silk proteins from other Araneoidea species, but not on their proline content. Our findings shed new light on the structural components of spider dragline silk, allowing further elucidation of their assembly properties, which may open the door for commercial applications.     

Silk elasticity as a potential constraint on spider body size

Silk is known for its strength and extensibility and has played a key role in the radiation of spiders. Individual spiders use different glands to produce silk types with unique sets of proteins. Most research has studied the properties of major ampullate and capture spiral silks and their ecological implications, while little is known about minor ampullate silk, the type used by those spider species studied to date for bridging displacements. A biomechanical model parameterised with available data shows that the minimum radius of silk filaments required for efficient bridging grows with the square root of the spider’s body mass, faster than the radius of minor ampullate silk filaments actually produced by spiders. Because the morphology of spiders adapted to walking along or under silk threads is ill suited for moving on a solid surface, for these species there is a negative relationship between body mass and displacement ability. As it stands, the model suggests that spiders that use silk for their displacements are prevented from attaining a large body size if they must track their resources in space. In particular, silk elasticity would favour sexual size dimorphism because males that must use bridging lines to search for females cannot grow large.