Changes in the material properties of spider glue droplet proteins accompanied shifts in prey capture biomechanics as cobweb spiders diverged from their orb weaving ancestors.

Although descended from orb weavers, spiders in the family Theridiidae spin cobwebs whose sticky prey capture gumfoot lines extend from a silk tangle to a surface below. When a crawling insect contacts glue droplets at the bottom of a gumfoot line, the line's weak pyriform anchor releases, causing the taut line to contract, pulling the insect from the surface and making its struggles to escape ineffective. To determine if this change in prey capture biomechanics was accompanied by a change in the material properties of theridiid glue, we characterized the elastic modulus and toughness of the glue droplet proteins of four theridiid species at 20-90 % relative humidity and compared their properties with those of 13 orb weaving species in the families Tetragnathidae and Araneidae. Compared to orb weavers, theridiid glue proteins had low extensions per protein volume and low elastic modulus and toughness values. These differences are likely explained by the loss of tension on a gumfoot line when its anchor fails, which may prioritize glue droplet adhesion rather than extension. Similarities in theridiid glue droplet properties did not reflect these species' evolutionary relationships. Instead, they appear associated with differences in web architecture. Two species that had stiffer gumfoot support lines and longer and more closely spaced gumfoot lines also had stiffer glue proteins. These lines may store more energy, and, when their anchors release, require stiffer glue to resist the more forceful upward thrust of a prey. STATEMENT OF SIGNIFICANCE: When a crawling insect contacts glue droplets on a theridiid cobweb's gumfoot line, this taut line's anchor fails and the insect is hoisted upward, rendering its struggles to escape ineffective. This strategy contrasts with that of orb weaving ancestors, which rely on more closely spaced prey capture threads to intercept and retain flying insects. A comparison of the elastic modulus and toughness of gumfoot and orb web glue proteins shows that this change in prey capture biomechanics is associated with reductions in the stiffness and toughness of cobweb glue. Unlike orb web capture threads, whose droplets extend in a coordinated fashion to sum adhesive forces, gumfoot lines become untethered, which prioritizes glue droplet adhesive contact over glue droplet extension.

Adhesive contact and protein elastic modulus tune orb weaving spider glue droplet biomechanics to habitat humidity.

Tiny glue droplets along the viscous capture threads of spider orb webs prevent insects from escaping. Each droplet is formed of a protein core surrounded by a hygroscopic aqueous layer, which cause the droplet's adhesion to change with humidity. As an insect struggles to escape the web, a thread's viscoelastic core proteins extend, transferring adhesive forces to the thread's support fibers. Maximum adhesive force is achieved when absorbed atmospheric moisture allows a flattened droplet to establish sufficient adhesive contact while maintaining the core protein cohesion necessary for force transfer. We examined the relationship between these droplet properties and adhesive force and the work of extending droplets at five relative humidities in twelve species that occupy habitats which have different humidities. A regression analysis that included both flattened droplet area and core protein elastic modulus described droplet adhesion, but the model was degraded when core protein area was substituted for droplet. Species from low humidity habitats expressed greater adhesion at lower humidities, whereas species from high humidity habitats expressed greater adhesion at high humidities. Our results suggest a general model of droplet adhesion with two adhesion peaks, one for low humidity species, which occurs when increasing droplet area and decreasing protein cohesion intersect, and another for high humidity species, which occurs when area and cohesion have diverged maximally. These dual peaks in adhesive force explain why some species from intermediate and high humidity habitats express high adhesion at several humidities. STATEMENT OF SIGNIFICANCE: We characterized the effect of humidity on the adhesion of twelve orb weaving spider species' glue droplets and showed how humidity-mediated changes in the contact area of a droplet's outer, hygroscopic aqueous layer and the stiffness of its protein core affect droplet performance. This revealed how droplet adhesion has been tuned to the humidity of a species' habitat and allowed us to revise a model that describes the environmental determinants of droplet biomechanics.

Correlated evolution between orb weaver glue droplets and supporting fibres maintains their distinct biomechanical roles in adhesion.

Orb weaving spiders employ a 'silken toolkit' to accomplish a range of tasks, including retaining prey that strike their webs. This is accomplished by a viscous capture spiral thread that features tiny glue droplets, supported by a pair of elastic flagelliform fibres. Each droplet contains a glycoprotein core responsible for adhesion. However, prey retention relies on the integrated performance of multiple glue droplets and their supporting fibres, with previous studies demonstrating that a suspension bridge forms, whose biomechanics sum the adhesive forces of multiple droplets while dissipating the energy of the struggling insect. While the interdependence of the droplet's glycoprotein and flagelliform fibres for functional adhesion is acknowledged, there has been no direct test of this hypothesized linkage between the material properties of each component. Spider mass, which differs greatly across orb weaving species, also has the potential to affect flagelliform fibre and glycoprotein material properties. Previous studies have linked spider mass to capture thread performance but have not examined the relationship between spider mass and thread material properties. We extend earlier studies to examine these relationships in 16 orb weaving species using phylogenetic generalized least squares. This analysis revealed that glycoprotein stiffness (elastic modulus) was correlated with flagelliform fibre stiffness, and that spider mass was related to the glycoprotein volume, flagelliform fibre cross-sectional area and droplets per unit thread length. By shaping the elastic moduli of glycoprotein adhesive and flagelliform fibres, natural selection has maintained the biomechanical integration of this adhesive system.

Humidity mediated performance and material properties of orb weaving spider adhesive droplets.

Capture thread glue droplets retain insects that strike an orb web and are key to the success of over 4,600 described spider species. Each droplet is a self-assembling adhesive system whose emergent biomechanical properties are centered on its viscoelastic, protein core. This bioadhesive is dependent on its surrounding hygroscopic aqueous layer for hydration and chemical conditioning. Consequently, a droplet's water content and adhesive performance track environmental humidity. We tested the hypothesis that natural selection has tuned a droplet's adhesive performance and material properties to a species' foraging humidity. At 55% relative humidity (RH) the adhesive properties of 12 species ranged from that of PEG-based hydrogels to that of silicone rubber, exhibiting a 1088-fold inter-specific difference in stiffness (0.02-21.76 MPa) and a 147-fold difference in toughness (0.14-20.51 MJ/m3). When tested over a 70% RH range, droplet extension lengths per protein core volume peaked at lower humidities in species from exposed, low humidity habitats, and at higher humidities in nocturnal species and those found in humid habitats. However, at the RH's where these species' maximum extension per protein volume indices were observed, the stiffness of most species' adhesive did not differ, documenting that selection has tuned elastic modulus by adjusting droplet hygroscopicity. This inverse relationship between droplet hygroscopicity and a species' foraging humidity ensures optimal adhesive stiffness. By characterizing the humidity responsiveness and properties of orb spider glue droplets, our study also profiles the range of its biomimetic potential. STATEMENT OF SIGNIFICANCE: Over 4,600 described species of orb weaving spider rely on tiny glue droplets in their webs to retain insect that the web intercepts. The aqueous layer that covers each droplet's core allows this adhesive to remain pliable and to stretch as an insect struggles to escape. The aqueous solution also attracts water from the air, causing the glue droplet's performance to change with humidity. By characterizing the droplet extensions and adhesive material properties of twelve species at relative humidities between of 20 and 90%, this study examined how this unique adhesive system responds to its environment and how it is tuned to the humidity of a species' habitat.

Protein Composition and Associated Material Properties of Cobweb Spiders' Gumfoot Glue Droplets.

The origin of aggregate silk glands and their production of wet adhesive silks is considered a key innovation of the Araneoidea, a superfamily of spiders that build orb-webs and cobwebs. Orb-web weavers place aggregate glue on an extensible capture spiral, whereas cobweb weavers add it to the ends of strong, stiff fibers, called gumfoot lines. Here we describe the material behavior and quantitative proteomics of the aggregate glues of two cobweb weaving species, the western black widow, Latrodectus hesperus, and the common house spider, Parasteatoda tepidariorum. For each species, respectively, we identified 48 and 33 proteins that were significantly more abundant in the portion of the gumfoot line with glue than in its fibers. These proteins were more highly glycosylated and phosphorylated than proteins found in silk fibers without glue, which likely explains aggregate glue stickiness. Most glue-enriched proteins were of anterior aggregate gland origin, supporting the hypothesis that cobweb weavers' posterior aggregate glue is specialized for another function. We found that cobweb weaver glue droplets are stiffer and tougher than the adhesive of most orb-web weaving species. Attributes of gumfoot glue protein composition that likely contribute to this stiffness include the presence of multiple protein families with conserved cysteine residues, a bimodal distribution of isoelectric points, and families with conserved functions in protein aggregation, all of which should contribute to cohesive protein-protein interactions. House spider aggregate droplets were more responsive to humidity changes than black widow droplets, which could be mediated by differences in protein sequence, post-translational modifications, the non-protein components of the glue droplets, and/or the larger amount of aqueous material that surrounds the adhesive cores of their glue droplets.

Linking properties of an orb-weaving spider's capture thread glycoprotein adhesive and flagelliform fiber components to prey retention time.

An orb web's adhesive capture spiral is responsible for prey retention. This thread is formed of regularly spaced glue droplets supported by two flagelliform axial lines. Each glue droplet features a glycoprotein adhesive core covered by a hygroscopic aqueous layer, which also covers axial lines between the droplets, making the entire thread responsive to environmental humidity.We characterized the effect of relative humidity (RH) on ability of Argiope aurantia and Argiope trifasciata thread arrays to retain houseflies and characterize the effect of humidity on their droplet properties. Using these data and those of Araneus marmoreus from a previous study, we then develop a regression model that correlated glycoprotein and flagelliform fiber properties with prey retention time. The model selection process included newly determined, humidity-specific Young's modulus and toughness values for the three species' glycoproteins.Argiope aurantia droplets are more hygroscopic than A. trifasciata droplets, causing the glycoprotein within A. aurantia droplets to become oversaturated at RH greater than 55% RH and their extension to decrease, whereas A. trifasciata droplet performance increases to 72% RH. This difference is reflected in species' prey retention times, with that of A. aurantia peaking at 55% RH and that of A. trifasciata at 72% RH.Fly retention time was explained by a regression model of five variables: glue droplet distribution, flagelliform fiber work of extension, glycoprotein volume, glycoprotein thickness, and glycoprotein Young's modulus.The material properties of both glycoprotein and flagelliform fibers appear to be phylogenetically constrained, whereas natural selection can more freely act on the amount of each material invested in a thread and on components of the thread's aqueous layer. Thus, it becomes easier to understand how natural selection can tune the performance of viscous capture threads by directing small changes in these components.

Orb weaver glycoprotein is a smart biological material, capable of repeated adhesion cycles.

Orb weavers produce webs that trap prey using a capture spiral formed of regularly spaced glue droplets supported by protein fibers. Each droplet consists of an outer aqueous layer and an adhesive, viscoelastic glycoprotein core. Organic and inorganic compounds in the aqueous layer make droplets hygroscopic and cause droplet features to change with environmental humidity. When droplets contact a surface, they adhere and extend as an insect struggles. Thus, a droplet's extensibility is as important for prey capture as its adhesion. Cursory observations show that droplets can adhere, extend, and pull off from a surface several times, a process called cycling. Our study cycled individual droplets of four species-Argiope aurantia, Neoscona crucifera, Verrucosa arenata, and Larinioides cornutus. Droplets were subjected to 40 cycles at two humidities to determine how humidity affected droplet performance. We hypothesized that droplets would continue to perform, but that performance would decrease. Droplet performance was characterized by filament length and force on droplets at pull-off, aqueous volume, and glycoprotein volume. As hypothesized, cycling decreased performance, notably extensibility and aqueous volume. However, humidity did not impact the response to cycling. In a natural context, droplets are not subjected to extensive cycling, but reusability is advantageous for orb-weaving spiders. Moreover, the ability to cycle, combined with their environmental responsiveness, allows us to characterize orb weaver droplets as smart materials for the first time.

Properties of orb weaving spider glycoprotein glue change during Argiope trifasciata web construction.

An orb web's prey capture thread relies on its glue droplets to retain insects until a spider can subdue them. Each droplet's viscoelastic glycoprotein adhesive core extends to dissipate the forces of prey struggle as it transfers force to stiffer, support line flagelliform fibers. In large orb webs, switchback capture thread turns are placed at the bottom of the web before a continuous capture spiral progresses from the web's periphery to its interior. To determine if the properties of capture thread droplets change during web spinning, we characterized droplet and glycoprotein volumes and material properties from the bottom, top, middle, and inner regions of webs. Both droplet and glycoprotein volume decreased during web construction, but there was a progressive increase in the glycoprotein's Young's modulus and toughness. Increases in the percentage of droplet aqueous material indicated that these increases in material properties are not due to reduced glycoprotein viscosity resulting from lower droplet hygroscopicity. Instead, they may result from changes in aqueous layer compounds that condition the glycoprotein. A 6-fold difference in glycoprotein toughness and a 70-fold difference in Young's modulus across a web documents the phenotypic plasticity of this natural adhesive and its potential to inspire new materials.

Elastic modulus and toughness of orb spider glycoprotein glue.

An orb web's prey capture thread features tiny glue droplets, each formed of an adhesive glycoprotein core surrounded by an aqueous layer. Small molecules in the aqueous layer confer droplet hygroscopicity and maintain glycoprotein viscoelasticity, causing droplet volume and glycoprotein performance to track changes in environmental humidity. Droplet extension combines with that of a thread's supporting flagelliform fibers to sum the adhesive forces of multiple droplets, creating an effective adhesive system. We combined measurements of the force on an extending droplet, as gauged by the deflection of its support line, with measurements of glycoprotein volume and droplet extension to determine the Young's modulus (E) and toughness of three species' glycoproteins. We did this at five relative humidities between 20-90% to assess the effect of humidity on these properties. When droplets of a thread span extend, their extensions are constrained and their glycoprotein filaments remain covered by aqueous material. This was also the case during the first extension phase of the individual droplets that we examined. However, as extension progressed, the aqueous layer was progresses disrupted, exposing the glycoprotein. During the first extension phase E ranged from 0.00003 GPa, a value similar to that of fibronectin, a glycoprotein that anchors cells in the extracellular matrix, to 0.00292 GPa, a value similar to that of resilin in insect ligaments. Second phase E increased 4.7-19.4-fold. When compared at the same humidity the E of each species' glycoprotein was less than 5% of the value reported for its flagelliform fibers. This difference may facilitate the coordinated extension of these two capture thread components that is responsible for summing the thread's adhesive forces.

Tuning orb spider glycoprotein glue performance to habitat humidity.

Orb-weaving spiders use adhesive threads to delay the escape of insects from their webs until the spiders can locate and subdue the insects. These viscous threads are spun as paired flagelliform axial fibers coated by a cylinder of solution derived from the aggregate glands. As low molecular mass compounds (LMMCs) in the aggregate solution attract atmospheric moisture, the enlarging cylinder becomes unstable and divides into droplets. Within each droplet an adhesive glycoprotein core condenses. The plasticity and axial line extensibility of the glycoproteins are maintained by hygroscopic LMMCs. These compounds cause droplet volume to track changes in humidity and glycoprotein viscosity to vary approximately 1000-fold over the course of a day. Natural selection has tuned the performance of glycoprotein cores to the humidity of a species' foraging environment by altering the composition of its LMMCs. Thus, species from low-humidity habits have more hygroscopic threads than those from humid forests. However, at their respective foraging humidities, these species' glycoproteins have remarkably similar viscosities, ensuring optimal droplet adhesion by balancing glycoprotein adhesion and cohesion. Optimal viscosity is also essential for integrating the adhesion force of multiple droplets. As force is transferred to a thread's support line, extending droplets draw it into a parabolic configuration, implementing a suspension bridge mechanism that sums the adhesive force generated over the thread span. Thus, viscous capture threads extend an orb spider's phenotype as a highly integrated complex of large proteins and small molecules that function as a self-assembling, highly tuned, environmentally responsive, adhesive biomaterial. Understanding the synergistic role of chemistry and design in spider adhesives, particularly the ability to stick in wet conditions, provides insight in designing synthetic adhesives for biomedical applications.

Humidity-mediated changes in an orb spider's glycoprotein adhesive impact prey retention time.

Properties of the viscous prey capture threads of araneoid orb spiders change in response to their environment. Relative humidity (RH) affects the performance of the thread's hygroscopic droplets by altering the viscoelasticity of each droplet's adhesive glycoprotein core. Studies that have characterized this performance used smooth glass and steel surfaces and uniform forces. In this study, we tested the hypothesis that these changes in performance translate into differences in prey retention times. We first characterized the glycoprotein contact surface areas and maximum extension lengths of Araneus marmoreus droplets at 20%, 37%, 55%, 72% and 90% RH and then modeled the relative work required to initiate pull-off of a 4 mm thread span, concluding that this species' droplets and threads performed optimally at 72% RH. Next, we evaluated the ability of three equally spaced capture thread strands to retain a house fly at 37%, 55% and 72% RH. Each fly's struggle was captured in a video and bouts of active escape behavior were summed. House flies were retained 11 s longer at 72% RH than at 37% and 55% RH. This difference is ecologically significant because the short time after an insect strikes a web and before a spider begins wrapping it is an insect's only opportunity to escape from the web. Moreover, these results validate the mechanism by which natural selection can tune the performance of an orb spider's capture threads to the humidity of its habitat.

Around the World in Eight Million Years: Historical Biogeography and Evolution of the Spray Zone Spider Amaurobioides (Araneae: Anyphaenidae).

Closely related organisms with transoceanic distributions have long been the focus of historical biogeography, prompting the question of whether long-distance dispersal, or tectonic-driven vicariance shaped their current distribution. Regarding the Southern Hemisphere continents, this question deals with the break-up of the Gondwanan landmass, which has also affected global wind and oceanic current patterns since the Miocene. With the advent of phylogenetic node age estimation and parametric bioinformatic advances, researchers have been able to disentangle historical evolutionary processes of taxa with greater accuracy. In this study, we used the coastal spider genus Amaurobioides to investigate the historical biogeographical and evolutionary processes that shaped the modern-day distribution of species of this exceptional genus of spiders. As the only genus of the subfamily Amaurobioidinae found on three Southern Hemisphere continents, its distribution is well-suited to study in the context of Gondwanic vicariance versus long-distance, transoceanic dispersal. Ancestral species of the genus Amaurobioides appear to have undergone several long-distance dispersal events followed by successful establishments and speciation, starting from the mid-Miocene through to the Pleistocene. The most recent common ancestor of all present-day Amaurobioides species is estimated to have originated in Africa after arriving from South America during the Miocene. From Africa the subsequent dispersals are likely to have taken place predominantly in an eastward direction. The long-distance dispersal events by Amaurobioides mostly involved transoceanic crossings, which we propose occurred by rafting, aided by the Antarctic Circumpolar Current and the West Wind Drift.

Spiders Tune Glue Viscosity to Maximize Adhesion.

Adhesion in humid conditions is a fundamental challenge to both natural and synthetic adhesives. Yet, glue from most spider species becomes stickier as humidity increases. We find the adhesion of spider glue, from five diverse spider species, maximizes at very different humidities that matches their foraging habitats. By using high-speed imaging and spreading power law, we find that the glue viscosity varies over 5 orders of magnitude with humidity for each species, yet the viscosity at maximal adhesion for each species is nearly identical, 10(5)-10(6) cP. Many natural systems take advantage of viscosity to improve functional response, but spider glue's humidity responsiveness is a novel adaptation that makes the glue stickiest in each species' preferred habitat. This tuning is achieved by a combination of proteins and hygroscopic organic salts that determines water uptake in the glue. We therefore anticipate that manipulation of polymer-salts interaction to control viscosity can provide a simple mechanism to design humidity responsive smart adhesives.

The impact of UVB radiation on the glycoprotein glue of orb-weaving spider capture thread.

Many spider orb-webs are exposed to sunlight and the potentially damaging effects of ultraviolet B (UVB) radiation. We examined the effect of UVB on the viscoelastic glycoprotein core of glue droplets deposited on the prey capture threads of these webs, hypothesizing that webs built by species that occupy sunny habitats are less susceptible to UVB damage than are webs built by species that prefer shaded forest habitats or by nocturnal species. Threads were tested shortly after being collected in the early morning and after being exposed to UVB energy equivalent to a day of summer sun and three times this amount. Droplets kept in a dark chamber allowed us to evaluate post-production changes. Droplet volume was unaffected by treatments, indicating that UVB did not damage the hygroscopic compounds in the aqueous layer that covers droplets. UVB exposure did not affect energies of droplet extension for species from exposed and partially to mostly shaded habitats (Argiope aurantia, Leucauge venusta and Verrucosa arenata). However, UVB exposure reduced the energy of droplet extension in Micrathena gracilis from shaded forests and Neoscona crucifera, which forages at night. Only in L. venusta did the energy of droplet extension increase after the dark treatment, suggesting endogenous molecular alignment. This study adds UVB irradiation to the list of factors (humidity, temperature and strain rate) known to affect the performance of spider glycoprotein glue, factors that must be more fully understood if adhesives that mimic spider glycoprotein glue are to be produced.

Temperature mediates the effect of humidity on the viscoelasticity of glycoprotein glue within the droplets of an orb-weaving spider's prey capture threads.

Sticky viscous prey capture threads retain insects that strike araneoid orb-webs. The threads' two axial fibers support a series of glue droplets, each featuring a core of adhesive viscoelastic glycoprotein covered by an aqueous solution. After sticking, the glue extends, summing the adhesion of multiple droplets, and dissipates some of the energy of a struggling prey. As a day progresses, threads experience a drop in humidity and an increase in temperature, environmental variables that have the potential to alter thread and web function. We hypothesize that thread droplets respond to these opposing environmental changes in a manner that stabilizes their performance, and test this by examining threads spun by Argiope aurantia, a species that occupies exposed, weedy habitats. We confirmed that decreased humidity increases glycoprotein viscosity and found that increased temperature had the opposite effect. To evaluate the combined effect of temperature and humidity on a droplet's ability to transfer adhesive force and dissipate energy, we extended a droplet and measured both the deflection of the axial line supporting the droplet and the duration of its tensive load. The cumulative product of these two indices, which reflects the energy required to extend a droplet, was greatest under afternoon (hot and dry) conditions, less under morning (cool and humid) conditions, and least under hot and humid afternoon conditions. Although the opposing effects of temperature and humidity tend to stabilize glycoprotein performance, A. aurantia thread droplets appear to function optimally during the afternoon, equipping this species to capture large orthopterans, which are most active at this time.

Environmental response and adaptation of glycoprotein glue within the droplets of viscous prey capture threads from araneoid spider orb-webs.

Viscous threads that form the prey capture spiral of araneoid orb-webs retain insects that strike the web, giving a spider more time to locate and subdue them. The viscoelastic glycoprotein glue responsible for this adhesion forms the core of regularly spaced aqueous droplets, which are supported by protein axial fibers. Glycoprotein extensibility both facilitates the recruitment of adhesion from multiple droplets and dissipates the energy generated by insects struggling to free themselves from the web. Compounds in the aqueous material make the droplets hygroscopic, causing an increase in both droplet volume and extensibility as humidity (RH) rises. We characterized these humidity-mediated responses at 20%, 37%, 55%, 72% and 90% RH in two large orb-weavers, Argiope aurantia, which is found in exposed habitats, and Neoscona crucifera, which occupies forests and forest edges. The volume-specific extension of A. aurantia glycoprotein reached a maximum value at 55% RH and then declined, whereas that of N. crucifera increased exponentially through the RH range. As RH increased, the relative stress on droplet filaments at maximum extension, as gauged by axial line deflection, decreased in a linear fashion in A. aurantia, but in N. crucifer increased logarithmically, indicating that N. crucifera threads are better equipped to dissipate energy through droplet elongation. The greater hygroscopicity of A. aurantia threads equips them to function in lower RH environments and during the afternoon when RH drops, but their performance is diminished during the high RH of the morning hours. In contrast, the lower hygroscopicity of N. crucifera threads optimizes their performance for intermediate and high RH environments and during the night and morning. These interspecific differences support the hypothesis that viscous capture threads are adapted to the humidity regime of an orb-weaver's habitat.

Humidity affects the extensibility of an orb-weaving spider's viscous thread droplets.

The prey-capture threads found in most spider orb webs rely on viscous droplets for their stickiness. Each droplet is formed of a central mass of viscoelastic glycoprotein glue surrounded by an aqueous covering, both of which incorporate hydrophilic components. We found that the extensibility of droplets on Larinioides cornutus threads increased as humidity increased. However, the deflection of the droplets' supporting axial lines did not change, indicating that atmospheric water uptake increases glycoprotein plasticity, but not glycoprotein adhesion. The extensibility of droplets, along with that of the thread's supporting axial fibers, is responsible for summing the adhesion of multiple thread droplets. Therefore, daily changes in humidity have the potential to significantly alter the performance of viscous threads and orb webs.

Constraints on the adhesion of viscous threads spun by orb-weaving spiders: the tensile strength of glycoprotein glue exceeds its adhesion.

In this study we tested the hypothesis that a viscous thread releases its hold on a surface because its glycoprotein glue pulls from the surface and not because its elongating droplets break near their attachment to the surface. We compared the values obtained when three species' viscous threads adhered to four smooth surfaces, which differed in their total surface energy and in the proportions of their dispersion and polar energy components. Although water comprised 43-70% of the volume of these viscous droplets, only the dispersion surface energies of test materials and not their polar surface energies impacted thread adhesion. These results support the droplet pull-off hypothesis and are consistent with a previous finding that capillary force contributes little to thread adhesion. Just as a viscous thread's stickiness is constrained by the tensile strength of its supporting axial fibers, our findings suggest that glycoprotein adhesion is constrained by glycoprotein tensile strength.

Adhesive compatibility of cribellar and viscous prey capture threads and its implication for the evolution of orb-weaving spiders.

Evolution of orb-weaving spiders that comprise the Orbiculariae clade involved a transition in the composition of prey capture thread that has been challenging to explain. The primitive cribellar threads spun by members of the Deinopoidea subclade resemble the capture threads of their non-orb-web-weaving ancestors and are formed of thousands of fine, dry, protein cribellar fibrils. In contrast, the derived viscous capture threads spun by members of the Araneoidea subclade have regularly spaced, aqueous adhesive droplets. When second instar deinopoid spiderlings emerge from egg sacs they are unable to spin cribellar threads, and, therefore, do not construct orb-webs; whereas second instar araneoids spin capture threads and construct orb-webs. If, as we hypothesize, viscous material evolved to enable second instar spiderlings to construct orb-webs, early araneoids may have spun composite cribellar-viscous capture threads. To examine the functional feasibility of such intermediate capture threads, we compared the adhesion of cribellar threads, viscous threads, and combined cribellar-viscous threads. The stickiness of these combined threads was greater than that of native cribellar or viscous threads alone. The viscous material of Araneus marmoreus threads exhibited a substantial increase in stickiness when combined with cribellar fibrils and that of Argiope aurantia threads a small increase in stickiness when combined with cribellar fibrils. Thus, if early araneoids retained their ability to spin cribellar threads after having evolved glands that produced viscous material, their composite threads could have formed a functional adhesive system that achieved its stickiness at no loss of material economy.

The adhesive delivery system of viscous capture threads spun by orb-weaving spiders.

The sticky viscous capture threads in araneoid orb-webs are responsible for retaining insects that strike these webs. We used features of 16 species' threads and the stickiness that they expressed on contact plates of four widths to model their adhesive delivery systems. Our results confirm that droplets at the edges of thread contact contribute the greatest adhesion, with each successively interior droplet contributing only 0.70 as much adhesion. Thus, regardless of the size and spacing of a thread's large primary droplets, little adhesion accrues beyond a span of 20 droplets. From this pattern we computed effective droplet number (EDN), an index that describes the total droplet equivalents that contribute to the stickiness of thread spans. EDN makes the greatest positive contribution to thread stickiness, followed by an index of the shape and size of primary droplets, and the volume of small secondary droplets. The proportion of water in droplets makes the single greatest negative contribution to thread stickiness, followed by a thread's extensibility, and the area of flattened droplets. Although highly significant, this six-variable model failed to convincingly describe the stickiness of six species, a problem resolved when species were assigned to three groups and a separate model was constructed for each. These models place different weights on the variables and, in some cases, reverse or exclude the contribution of a variable. Differences in threads may adapt them to particular habitats, web architectures or prey types, or they may be shaped by a species' phylogeny or metabolic capabilities.