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PubMed1998

Quantifying the venom dose of the spider Cupiennius salei using monoclonal antibodies.

The variation in venom dose with prey size of the neotropical wandering spider Cupieinnius salei was examined experimentally. Monoclonal antibodies were raised against the venom toxins of C. salei. Mab 9H3, recognizing the main toxin CSTX-1, was used to quantify the venom by enzyme-linked immunosorbent assay (ELISA). Crickets (Achta domesticus) in four size classes were randomly offered to sixteen mature female spiders at 14d intervals. The prey items were removed from spiders five minutes after the initial bite and subsequently homogenized for ELISA measurements. The quantity of venom expended was significantly related to the size of prey, ranging from 0.15 microl for the smallest (100 110 mg) to 1.53 microl for the largest (600-660 mg) crickets. Adaptations to prey size were also reflected in capturing behavior. None of the smallest, but almost 50% of the largest crickets were wrapped in silk following the spiders bite. Some other behavioral features may reduce the energetic costs of venom production. In 22% of the smallest crickets no venom was detectable, with the majority showing mechanical damage as a result of fang contact. This indicates. that C. salei does not rely exclusively on its venom when feeding on small prey. Some other aspects such as the site of the bite and the speed of paralyzation and their consequences associated with the amount of venom expended are discussed.

Animals↗
PubMed2000

Structural and ultrastructural description of the venom gland of Loxosceles intermedia (brown spider).

The brown spider, genus Loxosceles, is becoming of great medical importance, with envenomation (Loxoscelism) occurring throughout the world. The biological activities of the brown spider venom usually include dermonecrotic lesions at the bite site accompanied by hemolytic and haemorrhagic effects and also by renal failure. The objective of the present study was to describe the histology of the venom gland of L. intermedia using glands from adult spiders which were investigated by light microscopy, using immunohistochemical and staining methods, by transmission electron microscopy, and by scanning electron microscopy. The organization of the venom gland of Loxosceles intermedia follows the general architecture of spiders' venom glands. Using light microscopy and transmission electron microscopy we observed that the venom glands of L. intermedia present two layers of striated muscle fibers, an external layer and an internal layer in touch with an extracellular matrix which is a basement membrane structure and a fibrillar collagen matrix separating the muscular region from epithelial cells of the venom gland. Muscle cells are multinucleated, with nuclei peripherally placed and their cytoplasm rich in sarcoplasmic reticulum, myofibrills and continuous Z lines. By using scanning electron microscopy we can detect muscular cells from external layer as branching cells. Epithelial cells have their cytosol extremely rich in rough endoplasmic reticulum, mitochondria collection, Golgi apparatus, interdigitating membranes and secretory vesicles that ultimately accumulate the venom, a complex protein mixture.

Animals↗
PubMed1994

Dapsone or electric shock therapy of brown recluse spider envenomation?

STUDY OBJECTIVES: We tested the null hypothesis that no outcome differences exist among the treatment groups of animals with brown recluse spider envenomations. STUDY DESIGN: A prospective, placebo-controlled, experimental trial. Hartley guinea pigs were randomized into the following treatment groups: dapsone, Parali/azer stun gun, Guardian stun gun, and control. INTERVENTIONS: Brown recluse spider skin lesions were induced with intradermal injections of 30 micrograms spider venom and were treated beginning 16 hours after inoculation. Shock regimens consisted of four cross shocks of 1-second duration on anesthetized animals. Oral dapsone treatment was 0.7 mg/kg twice daily for 3 days. Lesion areas were measured daily for 3 days. RESULTS: The dapsone therapy group demonstrated significantly less induration and necrosis (P < .05) than that shown by the other three groups 72 hours after envenomation. CONCLUSION: Dapsone therapy is more effective than either electric shock or no therapy for brown recluse spider envenomation in the guinea pig model.

Analysis of Variance↗
PubMed2002

Blood phobia and spider phobia: two specific phobias with different autonomic cardiac modulations.

Cardiac reactions to two fear-related and one control film were compared in individuals high in spider or blood/injury fear. Twelve subjects in each phobic group were selected on the basis of their scores in the Spider or Mutilation Questionnaires and a semi-structured interview. Cardiac responses and self-reported affective ratings to the films were investigated. Sympathetic and parasympathetic cardiac influences were indexed by T-wave amplitude and respiratory sinus arrhythmia measured during film viewing. Basal parasympathetic cardiac control was also assessed during a paced breathing task. Results indicate differential autonomic modulation of cardiac responses for blood and spider phobics. Although each group reacted with marked cardiac activation to its feared stimulus, a sympathetic increase followed by withdrawal over time was found in blood phobics. Greater vagal tone at rest was present in blood phobics compared with spider phobics.

Adult↗
PubMed2003

From EST sequence to spider silk spinning: identification and molecular characterisation of Nephila senegalensis major ampullate gland peroxidase NsPox.

Spider dragline silk is renowned as one of the toughest materials of its kind. In nature, spider silks are spun out of aqueous solutions under environmental conditions. This is in contrast to production of most synthetic fibres, where hazardous solvents, high temperatures and pressure are used. In order to identify some of the chemical processes involved in spider silk spinning, we have produced a collection of cDNA sequences from specific regions of Nephila senegalensis major ampullate gland. We examined in detail the sequence and expression of a putative Nephila senegalensis peroxidase gene (NsPox) from our EST collection. NsPox encodes a protein with similarity to Drosophila melanogaster and Aedes aegypti peroxidases. Northern analysis and in situ localisation experiments revealed that NsPox is expressed in major and minor ampullate glands of the spider where the main components of the dragline silk are produced. We suggest that NsPox plays a role in dragline silk fibre formation and/or processing.

Amino Acid Sequence↗
PubMed2004

From a comb to a tree: phylogenetic relationships of the comb-footed spiders (Araneae, Theridiidae) inferred from nuclear and mitochondrial genes.

The family Theridiidae is one of the most diverse assemblages of spiders, from both a morphological and ecological point of view. The family includes some of the very few cases of sociality reported in spiders, in addition to bizarre foraging behaviors such as kleptoparasitism and araneophagy, and highly diverse web architecture. Theridiids are one of the seven largest families in the Araneae, with about 2200 species described. However, this species diversity is currently grouped in half the number of genera described for other spider families of similar species richness. Recent cladistic analyses of morphological data have provided an undeniable advance in identifying the closest relatives of the theridiids as well as establishing the family's monophyly. Nevertheless, the comb-footed spiders remain an assemblage of poorly defined genera, among which hypothesized relationships have yet to be examined thoroughly. Providing a robust cladistic structure for the Theridiidae is an essential step towards the clarification of the taxonomy of the group and the interpretation of the evolution of the diverse traits found in the family. Here we present results of a molecular phylogenetic analysis of a broad taxonomic sample of the family (40 taxa in 33 of the 79 currently recognized genera) and representatives of nine additional araneoid families, using approximately 2.5kb corresponding to fragments of three nuclear genes (Histone 3, 18SrDNA, and 28SrDNA) and two mitochondrial genes (16SrDNA and CoI). Several methods for incorporating indel information into the phylogenetic analysis are explored, and partition support for the different clades and sensitivity of the results to different assumptions of the analysis are examined as well. Our results marginally support theridiid monophyly, although the phylogenetic structure of the outgroup is unstable and largely contradicts current phylogenetic hypotheses based on morphological data. Several groups of theridiids receive strong support in most of the analyses: latrodectines, argyrodines, hadrotarsines, a revised version of spintharines and two clades including all theridiids without trace of a colulus and those without colular setae. However, the interrelationships of these clades are sensitive to data perturbations and changes in the analysis assumptions.

Animals↗
PubMed1995

Construction, cloning, and expression of synthetic genes encoding spider dragline silk.

Synthetic genes encoding recombinant spider silk proteins have been constructed, cloned, and expressed. Protein sequences were derived from Nephila clavipes dragline silk proteins and reverse-translated to the corresponding DNA sequences. Codon selection was chosen to maximize expression levels in Escherichia coli. DNA "monomer" sequences were multimerized to encode high molecular weight synthetic spider silks using a "head-to-tail" construction strategy. Multimers were cloned into a prokaryotic expression vector and the encoded silk proteins were expressed in E. coli upon induction with IPTG. Four multimer, ranging in size from 14.7 to 41.3 kDa, were chosen for detailed analysis. These proteins were isolated by immobilized metal affinity chromatography and purified using reverse-phase HPLC. The composition and identity of the purified proteins were confirmed by amino acid composition analysis, N-terminal sequencing, laser desorption mass spectroscopy, and Western analysis using antibodies reactive to native spider dragline silk. Circular dichroism measurements indicate that the synthetic spider silks have substantial beta-sheet structure.

Amino Acid Sequence↗
PubMed2004

Primary structure elements of spider dragline silks and their contribution to protein solubility.

Spider silk proteins have mainly been investigated with regard to their contribution to mechanical properties of the silk thread. However, little is known about the molecular mechanisms of silk assembly. As a first step toward characterizing this process, we aimed to identify primary structure elements of the garden spider's (Araneus diadematus) major dragline silk proteins ADF-3 and ADF-4 that determine protein solubility. In addition, we investigated the influence of conditions involved in mediating natural thread assembly on protein aggregation. Genes encoding spider silk-like proteins were generated using a cloning strategy, which is based on a combination of synthetic DNA modules and PCR-amplified authentic gene sequences. Comparing secondary structure, solubility, and aggregation properties of the synthesized proteins revealed that single primary structure elements have diverse influences on protein characteristics. Repetitive regions representing the largest part of dragline silk proteins determined the solubility of the synthetic proteins, which differed greatly between constructs derived from ADF-3 and ADF-4. Factors, such as acidification and increases in phosphate concentration, which promote silk assembly in vivo generally decreased silk protein solubility in vitro. Strikingly, this effect was pronounced in engineered proteins comprising the carboxyl-terminal nonrepetitive regions of ADF-3 or ADF-4, indicating that these regions might play an important role in initiating assembly of spider silk proteins.

Amino Acid Motifs↗
PubMedDate not supplied

Modeling of mechanical properties and structural design of spider web.

With a unique combination of strength and toughness among materials, spider silk is the model for engineering materials. This paper presents the stress-strain behavior of Nephila clavipes spider silk under tension, transverse compression, and torsional deformation obtained by a battery of micro testing equipment. The experimental results showed significantly higher toughness than the state-of-the-art fibers in tension and in transverse compression. Higher shear modulus was also observed for the spider silk comparing to other liquid crystalline fibers such as aramid fibers. On the basis of the experimental results finite element analysis is used to simulate static and dynamic properties of spider web and to explore the role of both material properties and architectural design in its structural integrity and mechanical performance.

Animals↗
PubMed2004

WISE NMR characterization of nanoscale heterogeneity and mobility in supercontracted Nephila clavipes spider dragline silk.

The addition of water to spider dragline silk results in fiber contraction to 50% its initial length and significant changes to the mechanical properties of the silk. This event has been termed supercontraction. A decrease in strength and increase in elasticity have been reported when the silk is in contact with water. Two-dimensional wide-line separation (WISE) nuclear magnetic resonance (NMR) is implemented to correlate (13)C chemical shifts with mobility by observing the corresponding (1)H line widths and line shapes in water-saturated spider dragline silk. The WISE NMR spectrum of the native silk exhibits (1)H line widths that are approximately 40 kHz for all carbon environments characteristic of a rigid organic system. In contrast, the water-saturated case displays a component of the (1)H line that is narrowed to approximately 5 kHz for the glycine C(alpha) and a newly resolved alanine helical environment while the alanine C(beta) corresponding to the beta-sheet conformation remains broad. These results indicate that water permeates the amorphous, glycine-rich matrix and not the crystalline, polyalanine beta-sheets. A delay time is added to the WISE NMR pulse sequence to monitor spin diffusion between the amorphous, mobile region and the crystalline domains. The time required for spin diffusion to reach spatial equilibrium is related to the length scale of the polyalanine crystallites. This technique is employed to measure crystalline domain sizes on the nanometer length scale in water-solvated spider dragline silk. These results provide further insight into the structure of spider silk and mechanism of supercontraction.

Amino Acid Sequence↗
PubMed2002

Mimicry of host cuticular hydrocarbons by salticid spider Cosmophasis bitaeniata that preys on larvae of tree ants Oecophylla smaragdina.

The salticid spider Cosmophasis bitaeniata preys on the larvae of the green tree ant Oecophylla smaragdina. Gas chromatography (GC) and gas chromatography-mass spectrometry (GC-MS) reveal that the cuticle of C. bitaeniata mimics the mono- and dimethylalkanes of the cuticle of its prey. Recognition bioassays with extracts of the cuticular hydrocarbons of ants and spiders revealed that foraging major workers did not respond aggressively to the extracts of the spiders or conspecific nestmates, but reacted aggressively to conspecific nonnestmates. Typically, the ants either failed to react (as with control treatments with no extracts) or they reacted nonaggressively as with conspecific nestmates. These data indicate that the qualitative chemical mimicry of ants by C. bitaeniata allows the spiders to avoid detection by major workers of O. smaragdina.

Aggression↗
PubMed2006

Phylo- and ontogenetic fears and the expectation of danger: differences between spider- and flight-phobic subjects in cognitive and physiological responses to disorder-specific stimuli.

To compare specific phobias with an assumed phylogenetic or ontogenetic origin in responses to fear-relevant (FR) stimuli, 17 spider- and 17 flight-phobic participants were exposed to pictures of spiders, flight accidents, or mushrooms randomly followed by either a startling noise or nothing else. While both groups showed a disorder-specific expectancy bias, only spider-phobic participants exhibited a disorder-specific covariation bias. Spider-phobic participants also showed enhanced skin conductance responses (SCRs), event-related brain potentials (ERPs), and startle responses triggered by disorder-specific FR pictures while flight-phobic participants showed only disorder-specific enhanced SCRs. In sum, our direct comparison between ontogenetic and phylogenetic phobias revealed that the former is characterized by biased and enhanced responses triggered by disorder-specific FR stimuli presumably based on a biological preparedness.

Adult↗
PubMed2003

Palaeontology: spider-web silk from the Early Cretaceous.

The use of viscid silk in aerial webs as a means to capture prey was a key innovation of araneoid spiders and has contributed largely to their ecological success. Here I describe a single silk thread from a spider's web that bears glue droplets and has been preserved in Lebanese amber from the Early Cretaceous period for about 130 million years. This specimen not only demonstrates the antiquity of viscid silk and of the spider superfamily Araneoidea, but is also some 90 million years older than the oldest viscid spider thread previously reported in Baltic amber from the Eocene epoch.

Adhesives↗
PubMed2003

Molecular nanosprings in spider capture-silk threads.

Spider capture silk is a natural material that outperforms almost any synthetic material in its combination of strength and elasticity. The structure of this remarkable material is still largely unknown, because spider-silk proteins have not been crystallized. Capture silk is the sticky spiral in the webs of orb-weaving spiders. Here we are investigating specifically the capture spiral threads from Araneus, an ecribellate orb-weaving spider. The major protein of these threads is flagelliform protein, a variety of silk fibroin. We present models for molecular and supramolecular structures of flagelliform protein, derived from amino acid sequences, force spectroscopy (molecular pulling) and stretching of bulk capture web. Pulling on molecules in capture-silk fibres from Araneus has revealed rupture peaks due to sacrificial bonds, characteristic of other self-healing biomaterials. The overall force changes are exponential for both capture-silk molecules and intact strands of capture silk.

Animals↗
PubMed2002

Amyloidogenic nature of spider silk.

In spiders soluble proteins are converted to form insoluble silk fibres, stronger than steel. The final fibre product has long been the subject of study; however, little is known about the conversion process in the silk-producing gland of the spider. Here we describe a study of the conversion of the soluble form of the major spider-silk protein, spidroin, directly extracted from the silk gland, to a beta-sheet enriched state using circular dichroism (CD) spectroscopy. Combined with electron microscopy (EM) data showing fibril formation in the beta-sheet rich region of the gland and amino-acid sequence analyses linking spidroin and amyloids, these results lead us to suggest that the refolding conversion is amyloid like. We also propose that spider silk could be a valuable model system for testing hypotheses concerning beta-sheet formation in other fibrilogenic systems, including amyloids.

Amino Acid Sequence↗
PubMed2002

Segmented nanofibers of spider dragline silk: atomic force microscopy and single-molecule force spectroscopy.

Despite its remarkable materials properties, the structure of spider dragline silk has remained unsolved. Results from two probe microscopy techniques provide new insights into the structure of spider dragline silk. A soluble synthetic protein from dragline silk spontaneously forms nanofibers, as observed by atomic force microscopy. These nanofibers have a segmented substructure. The segment length and amino acid sequence are consistent with a slab-like shape for individual silk protein molecules. The height and width of nanofiber segments suggest a stacking pattern of slab-like molecules in each nanofiber segment. This stacking pattern produces nano-crystals in an amorphous matrix, as observed previously by NMR and x-ray diffraction of spider dragline silk. The possible importance of nanofiber formation to native silk production is discussed. Force spectra for single molecules of the silk protein demonstrate that this protein unfolds through a number of rupture events, indicating a modular substructure within single silk protein molecules. A minimal unfolding module size is estimated to be around 14 nm, which corresponds to the extended length of a single repeated module, 38 amino acids long. The structure of this spider silk protein is distinctly different from the structures of other proteins that have been analyzed by single-molecule force spectroscopy, and the force spectra show correspondingly novel features.

Amino Acid Sequence↗
PubMed1991

Chemical attraction of kleptoparasitic flies to heteropteran insects caught by orb-weaving spiders.

Insects of the heteropteran families Pentatomidae (stink bugs) and Coreidae (squash bugs), when being eaten by the orb-weaving spider Nephila clavipes, attract flies of the family Milichiidae. The flies aggregate on the bugs and, as kleptoparasites, share in the spider's meal. Stink bugs and squash bugs typically eject defensive sprays when attacked; they do so when caught by Nephila, but the spray only minimally affects the spider. Evidence is presented indicating that it is the spray of the bugs that attracts milichiids to the spider's catch.

Aldehydes↗
PubMed1999

Horseshoe crab hemocyte-derived antimicrobial polypeptides, tachystatins, with sequence similarity to spider neurotoxins.

Antimicrobial peptides, named tachystatins A, B, and C, were identified from hemocytes of the horseshoe crab Tachypleus tridentatus. Tachystatins exhibited a broad spectrum of antimicrobial activity against Gram-negative and Gram-positive bacteria and fungi. Of these tachystatins, tachystatin C was most effective. Tachystatin A is homologous to tachystatin B, but tachystatin C has no significant sequence similarity to tachystatins A and B. Tachystatins A and B showed sequence similarity to omega-agatoxin-IVA of funnel web spider venom, a potent blocker of voltage-dependent calcium channels. However, they exhibited no blocking activity of the P-type calcium channel in rat Purkinje cells. Tachystatin C also showed sequence similarity to several insecticidal neurotoxins of spider venoms. Tachystatins A, B, and C bound significantly to chitin. A causal relationship was observed between chitin binding activity and antifungal activity. Tachystatins caused morphological changes against a budding yeast, and tachystatin C had a strong cell lysis activity. The septum between mother cell and bud, a chitin-rich region, was stained by fluorescence-labeled tachystatin C, suggesting that the primary recognizing substance on the cell wall is chitin. As horseshoe crab is a close relative of the spider, tachystatins and spider neurotoxins may have evolved from a common ancestral peptide, with adaptive functions.

Amino Acid Sequence↗