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Compared chemical properties of dermonecrotic and lethal toxins from spiders of the genus Loxosceles (Araneae).

Loxosceles spider venom usually causes a typical dermonecrotic lesion in bitten patients, but it may also cause systemic effects that may be lethal. Gel filtration on Sephadex G-100 of Loxosceles gaucho, L. laeta, or L. intermedia spider venoms resulted in three fractions (A, containing higher molecular mass components. B containing intermediate molecular mass components, and C with lower molecular mass components). The dermonecrotic and lethal activities were detected exclusively in fraction A of all three species. Analysis by SDS-PAGE showed that the major protein contained in fraction A has molecular weight approximately 35 kDa in L. gaucho and L. intermedia, but 32 kDa in L. laeta venom. These toxins were isolated from venoms of L. gaucho, L. laeta, and L. intermedia by SDS-PAGE followed by blotting to PVDF membrane and sequencing. A database search showed a high level of identity between each toxin and a fragment of the L. reclusa (North American spider) toxin. A multiple sequence alignment of the Loxosceles toxins showed many common identical residues in their N-terminal sequences. Identities ranged from 50.0% (L. gaucho and L. reclusa) to 61.1% (L. intermedia and L. reclusa). The purified toxins were also submitted to capillary electrophoresis peptide mapping after in situ partial hydrolysis of the blotted samples. The results obtained suggest that L. intermedia protein is more similar to L. laeta toxin than L. gaucho toxin and revealed a smaller homology between L. intermedia and L. gaucho. Altogether these findings suggest that the toxins responsible for most important activities of venoms of Loxosceles species have a molecular mass of 32-35 kDa and are probably homologous proteins.

Amino Acid Sequence↗

Web-building spiders attract prey by storing decaying matter.

The orb-weaving spider Nephila edulis incorporates into its web a band of decaying animal and plant matter. While earlier studies demonstrate that larger spiders utilise these debris bands as caches of food, the presence of plant matter suggests additional functions. When organic and plastic items were placed in the webs of N. edulis, some of the former but none of the latter were incorporated into the debris band. Using an Y-maze olfactometer, we show that sheep blowflies Lucilia cuprina are attracted to recently collected debris bands, but that this attraction does not persist over time. These data reveal an entirely novel foraging strategy, in which a sit-and-wait predator attracts insect prey by utilising the odours of decaying organic material. The spider's habit of replenishing the debris band may be necessary to maintain its efficacy for attracting prey.

Animals↗

Host selection by a kleptobiotic spider.

Why do kleptobiotic spiders of the genus Argyrodes seem to be associated with spiders of the genus Nephila worldwide? Observations following introduction of experimental insect prey of different sizes and weights on to host webs revealed that: (1) small prey are more effectively retained on the web of Nephila clavipes than on the web of another common host, Leucauge venusta. (2) N. clavipes did not consume small prey that accumulated on the web whereas larger, heavier prey were enveloped and stored. (3) We observed clear partitioning of prey items between N. clavipes and Argyrodes spp.; diet selection by Argyrodes did not overlap with that of N. clavipes but closely overlapped with that of L. venusta. (4) L. venusta responds very quickly to prey impact whereas N. clavipes is slower, offering a temporal window of opportunity for Argyrodes foraging. (5) The ability of L. venusta to detect and respond to small items also means that it acts aggressively to Argyrodes spp., whereas N. clavipes does not. Consequently, food-acquisition behaviours of Argyrodes were clearly less risky with N. clavipes compared with L. venusta. We conclude that when a kleptobiotic organism has a choice of various host species, it will opt for the least risky host that presents the highest rate of availability of food items. The fact that Nephila species present such characteristics explains the worldwide association with Argyrodes kleptobiotic spiders.

Animals↗

Evolution of satellite DNAs in a radiation of endemic Hawaiian spiders: does concerted evolution of highly repetitive sequences reflect evolutionary history?

Satellite DNAs are known for an unusual and nonuniform evolution characterized by rapid evolutionary change between species and concerted evolution leading to molecular homogeneity within species. In this paper we use satellite DNAs for phylogenetic analysis of a rapidly evolving lineage of spiders and compare the phylogeny with a hypothesis previously generated based on mitochondrial DNA and allozymes. The spiders examined include almost all species within a monophyletic clade of endemic Hawaiian Tetragnatha species, the spiny-leg clade. The phylogeny based on satellite sequences is largely congruent to those produced by mtDNA and allozymes, except that the satellite DNA yields much longer branches, with higher levels of support for any given node. Closely related species that have differentiated ecologically within an island are well resolved with satellite DNA but much less so with mtDNA. These results suggest that Tetragnatha stDNA repeats seem to be evolving gradually and cohesively during the diversification of these endemic Hawaiian spiders. The study also reveals gain-loss of satellite DNA copies during species diversification. We conclude that satellite DNA sequences may potentially be very useful for resolving relationships between rapidly evolving taxa within an adaptive radiation. In addition, satellite DNA as a nuclear marker suggests that hybridization or peripatry could play a possible role in species formation that cannot be revealed by mitochondrial markers due to its maternal inheritance.

Animals↗

Acetylcholine receptors in spider peripheral mechanosensilla.

Peripherally located parts of spider mechanosensory neurons are modulated by several neurotransmitters released from apposed efferent fibers. Activities of acetylcholine (ACh) synthesizing enzyme choline acetyltransferase (ChAT) and ACh degrading enzyme acetylcholine esterase (AChE) were previously found in some efferent fibers. ChAT activity was also present in all the mechanosensory neurons, while AChE activity was only found in some. We show that spider mechanosensory neurons and probably some efferent neurons are immunoreactive to a monoclonal antibody against muscarinic ACh receptors (mAChRs). However, application of muscarinic agonists did not change the physiological responses or membrane potentials of neurons in the lyriform organ VS-3. Similarly, the sensitivities of the neurons of trichobothria (filiform hairs) remained unchanged after application of these agonists. Therefore, activation of mAChRs may only modulate the function of spider mechanosensory neurons indirectly, for example, by affecting the release of other transmitter(s). However, a subgroup of VS-3 neurons was inhibited by ACh, which also depolarized the membrane similar to these neurons' responses to GABA, suggesting that ACh activates anion channels in these neurons. Interestingly, all of the neurons responding to ACh were the rapidly adapting Type A neurons that were previously shown to express AChE activity.

Acetylcholine↗

Measuring and quantifying dynamic visual signals in jumping spiders.

Animals emit visual signals that involve simultaneous, sequential movements of appendages that unfold with varying dynamics in time and space. Algorithms have been recently reported (e.g. Peters et al. in Anim Behav 64:131-146, 2002) that enable quantitative characterization of movements as optical flow patterns. For decades, acoustical signals have been rendered by techniques that decompose sound into amplitude, time, and spectral components. Using an optic-flow algorithm we examined visual courtship behaviours of jumping spiders and depict their complex visual signals as "speed waveform", "speed surface", and "speed waterfall" plots analogous to acoustic waveforms, spectrograms, and waterfall plots, respectively. In addition, these "speed profiles" are compatible with analytical techniques developed for auditory analysis. Using examples from the jumping spider Habronattus pugillis we show that we can statistically differentiate displays of different "sky island" populations supporting previous work on diversification. We also examined visual displays from the jumping spider Habronattus dossenus and show that distinct seismic components of vibratory displays are produced concurrently with statistically distinct motion signals. Given that dynamic visual signals are common, from insects to birds to mammals, we propose that optical-flow algorithms and the analyses described here will be useful for many researchers.

Algorithms↗

Expression patterns of leg genes in the mouthparts of the spider Cupiennius salei (Chelicerata: Arachnida).

The leg genes extradenticle, homothorax, dachshund, and Distal-less define three antagonistic developmental domains in the legs, but not in the antenna, of Drosophila. Here we report the expression patterns of these leg genes in the prosomal appendages of the spider Cupiennius salei. The prosoma of the spider bears six pairs of appendages: a pair of cheliceres, a pair of pedipalps, and four pairs of walking legs. Three types of appendages thus can be distinguished in the spider. We show here that in the pedipalp, the leg-like second prosomal appendage, the patterns are very similar to those in the legs themselves, indicating the presence of three antagonistic developmental domains in both appendage types. In contrast, in the chelicera, the fang-like first prosomal appendage, the patterns are different and there is no evidence for antagonistic domains. Together with data from Drosophila this suggests that leg-shaped morphology of arthropod appendages requires an underlying set of antagonistic developmental domains, whereas other morphologies (e.g. antenna, chelicera) may result from the loss of such antagonistic domains.

Animals↗

Double-stranded RNA interference in the spider Cupiennius salei: the role of Distal-less is evolutionarily conserved in arthropod appendage formation.

Chelicerates represent a basal arthropod group, which makes them an excellent system for the study of evolutionary processes in arthropods. To enable functional studies in chelicerates, we developed a double-stranded RNA-interference (RNAi) protocol for spiders while studying the function of the Distal-less gene. We isolated the Distal-less gene from the spider Cupiennius salei. Cs-Dll gene expression is first seen in cells of the prosomal segments before the outgrowth of the appendages. After the appendages have formed, Cs-Dll is expressed in the distal portion of the prosomal appendages, and in addition, in the labrum, in two pairs of opisthosmal (abdominal) limb buds, in the head region, and at the posterior-most end of the spider embryo. In embryos, in which Dll was silenced by RNAi, the distal part of the prosomal appendages was missing and the labrum was completely absent. Thus, Dll also plays a crucial role in labrum formation. However, the complete lack of labrum in RNAi embryos may point to a different nature of the labrum from the segmental appendages. Our data show that the expression of Dll in the appendages is conserved among arthropods, and furthermore that the role of Dll is evolutionarily conserved in the formation of segmental appendages in arthropods.

Animals↗

A Hox class 3 orthologue from the spider Cupiennius salei is expressed in a Hox-gene-like fashion.

The class 3 Hox gene orthologue in insects, zerknüllt (zen), is not expressed along the anterior-posterior axis, but only in extra-embryonic tissues, suggesting that it has lost its function as a normal Hox gene. To analyse whether this loss of Hox gene function has already occurred in a basal arthropod lineage, we have isolated a Hox3 orthologue from the spider Cupiennius salei. In contrast to the insect zen sequences, which have a highly diverged homeobox, the spider Hox3 gene orthologue, Cs-Hox3, shows a high sequence similarity to the class 3 Hox genes of other phyla, including chordates. In situ hybridization in early embryos shows that it is expressed in a continuous region covering the pedipalp segment and the four leg-bearing segments. This expression pattern suggests a Hox-gene-like function for Cs-Hox3. On the other hand, the expression pattern does not strictly follow the colinearity rule, as it overlaps fully with the expression domain of the class 1 orthologue of the spider, Cs-lab. Still, our data suggest that the ancestor of the arthropods must have had a class 3 Hox gene with a function in anterior-posterior axis specification and that this function has been lost in the lineage leading to the insects.

Amino Acid Sequence↗

Selection on male size, leg length and condition during mate search in a sexually highly dimorphic orb-weaving spider.

Mate search plays a central role in hypotheses for the adaptive significance of extreme female-biased sexual size dimorphism (SSD) in animals. Spiders (Araneae) are the only free-living terrestrial taxon where extreme SSD is common. The "gravity hypothesis" states that small body size in males is favoured during mate search in species where males have to climb to reach females, because body length is inversely proportional to achievable speed on vertical structures. However, locomotive performance of males may also depend on relative leg length. Here we examine selection on male body size and leg length during mate search in the highly dimorphic orb-weaving spider Argiope aurantia, using a multivariate approach to distinguish selection targeted at different components of size. Further, we investigate the scaling relationships between male size and energy reserves, and the differential loss of reserves. Adult males do not feed while roving, and a size-dependent differential energy storage capacity may thus affect male performance during mate search. Contrary to predictions, large body size was favoured in one of two populations, and this was due to selection for longer legs. Male size was not under selection in the second population, but we detected direct selection for longer third legs. Males lost energy reserves during mate search, but this was independent of male size and storage capacity scaled isometrically with size. Thus, mate search is unlikely to lead to selection for small male size, but the hypothesis that relatively longer legs in male spiders reflect a search-adapted morphology is supported.

Animals↗

Regenerated spider silk as a new biomaterial for MEMS.

Spider drag-line silk is introduced for the first time as a new biomaterial for Micro-Electro-Mechanical Systems (MEMS). The tasks accomplished in this paper were focused on mechanical characterization of regenerated spider silk under two conditions: (1) spin-coated thin film formed onto a silicon substrate; and (2) formation of a free-standing microbridge (800 x 800 x 40 microm3) obtained by a surface micromachining process. Micromechanical tests using a nano indentation machine showed the spider silk film having an elastic modulus of 7.3 GPa, a loss tangent of 0.044 and an UTS (Ultimate Tensile Strength) of 85.1 MPa.

Animals↗

An experimental study of spider-related covariation bias in 8- to 13-year-old children.

Covariation bias can be defined as phobic subjects' tendency to overestimate the association between phobic stimuli and aversive outcomes. The current study presents two experiments that examined this type of cognitive bias in children aged 8-13 years (N=147 in Experiment 1, N=240 in Experiment 2). Children completed a self-report questionnaire for measuring spider fear and then participated in a card game in which fear-relevant (i.e., spider) and fear-irrelevant (i.e., weapon and Pokémon) pictures were equally paired with negative and positive outcomes (respectively losing and winning candy). No evidence was found for a relationship between children's level of spider fear and the tendency to link negative consequences to fear-relevant pictures. Various methodological and theoretical explanations for this null finding are discussed.

Adolescent↗

Molecular diversification in spider venoms: a web of combinatorial peptide libraries.

Spider venoms are a rich source of novel pharmacologically and agrochemically interesting compounds that have received increased attention from pharmacologists and biochemists in recent years. The application of technologies derived from genomics and proteomics have led to the discovery of the enormous molecular diversity of those venoms, which consist mainly of peptides and proteins. The molecular diversity of spider peptides has been revealed by mass spectrometry and appears to be based on a limited set of structural scaffolds. Genetic analysis has led to a further understanding of the molecular evolution mechanisms presiding over the generation of these combinatorial peptide libraries. Gene duplication and focal hypermutation, which has been described in cone snails, appear to be common mechanisms to venomous mollusks and spiders. Post-translational modifications, fine structural variations and new molecular scaffolds are other potential mechanisms of toxin diversification, leading to the pharmacologically complex cocktails used for predation and defense.

Amino Acid Sequence↗

Molecular mechanism of spider silk elasticity.

Spider major ampullate (drag-line) silk is an extracellular fibrous protein which has impressive characteristics of strength and elasticity. This silk has been hypothesized to predominantly consist of a single protein, containing regions of antiparallel beta-sheets which are interspersed with amorphous segments responsible for its elastic properties. A rubber-like mechanism has been suggested to account for this elasticity, but the specific molecular mechanism is unknown. Using Fourier transform infrared spectroscopy (FTIR) we found evidence of either helix formation or reorientation of preexisting helices when axial tension is applied to the spider silk fiber. CD studies of a peptide derived from the silk gene repeat sequence show that it can form beta-sheets at high temperatures while alpha-helices are induced in 2,2,2-trifluoroethanol. These results suggest a possible molecular mechanism for the elasticity of spider silk fibers. It is proposed that the elastic process involves the formation and disruption of alpha-helical Ala-rich regions which are interspersed among stable beta-sheet domains.

Amino Acid Sequence↗

Fear of Spiders Questionnaire.

The Fear of Spiders Questionnaire (FSQ), an 18-item self-report questionnaire assessing spider phobia, was developed in an attempt to complement the information provided by the Spider Phobia Questionnaire (SPQ). Data obtained from 338 undergraduates revealed that the FSQ was able to discriminate phobics from nonphobics, and indicated decrements in phobic responding from pretest to posttest following cognitive therapy. Test-retest data, obtained from non-treatment control groups, indicated that scores on the FSQ are stable over a one month period. The FSQ also demonstrated adequate convergent validity due to its significant correlations with the SPQ and a behavioral avoidance test. Finally, a factor analysis revealed two factors accounting for 55% of the variance. It is argued that, compared to the SPQ, items on the FSQ are more explicit regarding the time period to be assessed, and may be more sensitive to differences between phobics and nonphobics and decrements in phobic responding following treatment.

Adolescent↗

Molecular action mechanism of spider toxin on glutamate receptor: role of 2,4-dihydroxyphenylacetic acid in toxin molecule.

Joro spider toxin (JSTX) isolated from Nephila clavata was shown to inhibit L-glutamate binding to rat brain synaptic membranes in a dose-dependent manner. 2,4-Dihydroxyphenylacetic acid (2,4-DHPA), a common moiety of spider toxins, also inhibited specifically L-glutamate binding at a concentration similar to that of the toxin. The binding activity inhibited by 2,4-DHPA or JSTX was recoverable on addition of ferric compound. These results suggest that 2,4-DHPA is a functional moiety in the toxin molecule and the biological action of spider toxin is explained by direct interaction with an Fe-S center which is known to play an important role for the glutamate binding.

Animals↗

A spider toxin (JSTX) inhibits L-glutamate uptake by rat brain synaptosomes.

Joro spider toxin (JSTX), a specific blocker of glutamate receptors, was found to exert a prominent suppressive action on the Na+-dependent binding of L-glutamate to synaptic membranes and on glutamate uptake by synaptosomes in a dose-dependent manner. In contrast, the synthesized 2,4-dihydroxyphenylacetylasparagine (2,4-DHPA-ASN), a common moiety of spider toxins, which has been shown to exhibit almost the same activity as intact JSTX with respect to the inhibition of Na+-independent glutamate binding to its synaptic membrane receptors, shows lower potency in inhibiting Na+-dependent binding and uptake of L-glutamate. From these findings, it is clear that JSTX has the ability to inhibit not only L-glutamate binding to its synaptic membrane receptors but also L-glutamate uptake by synaptosomes, and that polyamines linked to 2,4-DHPA-ASN in the molecule of spider toxins may participate in the inhibition of L-glutamate uptake.

Animals↗

Spider silkglands contain a tissue-specific alanine tRNA that accumulates in vitro in response to the stimulus for silk protein synthesis.

The large ampullate glands of the orb-web spider, Nephila clavipes provide massive amounts of fibroin throughout the lifetime of the adult female. We have developed methods to culture the glands and manipulate their biosynthetic activity. This has allowed us to monitor a series of molecular events that precede silk production in glands excised from appropriately stimulated animals. In this paper, we demonstrate that prior to the transient dramatic production of fibroin, such glands accumulate large amounts of tRNAs cognate to the abundant amino acids in spider silk. One of these, alanine tRNA, appears to consist of two isoaccepting forms--one constitutive, and the other silkgland specific. Moreover, the silkgland-specific form appears to accumulate preferentially in response to stimulation. This phenomenon of tissue-specific tRNA production appears similar to that found in the silkglands of Bombyx mori, but the spider system has the unique property of permitting manipulation in vitro. Thus, it provides an unusual opportunity to study the mechanism of regulated tRNA synthesis.

Animals↗