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Action of brown widow spider venom and botulinum toxin on the frog neuromuscular junction examined with the freeze-fracture technique.

1. Structural changes which normally accompany transmitter release at frog neuromuscular junctions are visualized with the freeze-fracture technique. The effects of brown widow spider venom and botulinum toxin were evaluated in terms of their ability to block or produce these structural changes. Changes produced by these neuropoisons were correlated with their known effects on neurotransmitter release. 3. Fusion of synaptic vesicles with the presynaptic plasmalemma, normally evoked by electrical stimulation, was abolished at neuromuscular junctions from frogs treated with botulinum toxin. 3. The concentration of large intramembranous particles in the presynaptic plasmalemma, an indication of the excess of synaptic vesicle fusion over recovery of synaptic vesicle membrane, was increased by treatment with brown widow spider venom, even in the presence of botulinum toxin. 4. When external calcium was present, sites of vesicle fusion induced by brown widow spider venom, as well as by electrical stimulation, were located mainly in the active zone. In the absence of external calcium, many plasmalemmal deformations, also though to be sites of vesicle fusion, were more evenly dispersed over the presynaptic surface of nerve terminals. 5. Botulinum toxin decreased the number of vesicle fusion sites in the active zone induced by spider venom in the presence of external calcium but had little effect on the number of fusion sites induced by spider venom in the absence of external calcium. 6. Nerve terminals soaked in a sodium-free Ringer solution were partially depleted of vesicles. Addition of spider venom to this Ringer did not cause additional depletion of vesicles. 7. Formation of cation-permeable channels in the presynaptic membrane could account for these effects of spider venom on the frog neuromuscular junction. Botulinum toxin blocks vesicle fusion by some means which is not yet understood.

Animals

Inactivation of complement by Loxosceles reclusa spider venom.

Zymosan depletion of serum complement in guinea pigs rendered them highly resistant to lesion by Loxosceles reclusa spider venom. Guinea pigs deficient in C4 of the complement system are as sensitive to the venom as normal guinea pigs. The injection of 35 micrograms of whole recluse venom intradermally into guinea pigs lowered their complement level by 35.7%. Brown recluse spider venom in concentrations as slight as 0.02 micrograms protein/ml can totally inactivate one CH50 of guinea pig complement in vitro. Bee, scorpion, and other spider venoms had no influence on the hemolytic titer of complement. Fractionation of recluse spider venom by Sephadex G-200 filtration separated the complement-inactivating property of the venom into three major regions which could be distinguished on the basis of heat stability as well as size. None was neutralized by antivenom. Polyacrylamide gel electrophoresis of venom resolved the complement inactivators into five fractions. Complement inactivated by whole venom or the Sephadex fractions could be restored to hemolytic activity by supplements of fresh serum but not by heat-inactivated serum, pure C3, pure C5, or C3 and C5 in combination.

Animals

Effect of concanavalin A on black widow spider venom activity at the neuromuscular junction: implications for mechanisms of venom action.

Concanavalin A (Con A) inhibits black widow spider venom-induced transmitter release at both tissue-cultured and adult neuromuscular junctions and also inhibits the venom-induced destruction of cultured neurites. This inhibitory action is partially or completely prevented by prior treatment with colchicine. Neither colchicine nor Con A interacts significantly with depolarization-induced transmitter release. These results are analogous to those obtained from experiments on lymphocyte surface receptor capping. They suggest that redistribution of neuronal membrane components may be a crucial step in spider venom action. This membrane redistribution appears to be modulated in neurons, as in other cell types, by microtubule-microfilament array. How such a redistribution causes increased transmitter release cannot as yet be specified. Changes in the ionic permeability of sodium and potassium were examined as likely mechanisms. Increased sodium influx (and seondary release of calcium from intracellular stores) cannot be the basis for spider venom action. Increased potassium efflux remains a possibility, but is not consistent with all of the data. Other possible mechanisms are also suggested.

Animals

Sex- and development-specific transcriptomic profiling of venom and silk genes in the wolf spider Pardosa astrigera provides insights into ecological adaptation and predatory strategies.

Spider venom and silk glands are two major secretory systems that contribute to prey capture, defense, and reproduction, but their sex- and development-specific molecular regulation in wandering wolf spiders remains poorly understood. Here, the transcriptome of Pardosa astrigera, an important agricultural natural enemy in China, revealed significant sex- and development-associated molecular differentiation among adult females, adult males, and spiderlings. A total of 100,025 unigenes were obtained, of which 23,852 were functionally annotated, providing a comprehensive transcriptomic resource for this species. Differential expression patterns showed marked variation among groups, with 531, 1792, and 832 DEGs detected in PAF vs PAS, PAM vs PAS, and PAF vs PAM, respectively. These genes were mainly associated with metabolic, oxidation-reduction, cuticle development, MAPK signaling, and lysosome pathways. Fifteen co-expression modules revealed distinct expression patterns. The turquoise, pink, yellow, and red modules were development-related, whereas the blue module was male-biased. Venom- and spidroin-related genes were distributed across multiple modules, suggesting coordinated regulation. Overall, 42 venom peptides, 21 venom proteins, and 11 spidroins were identified. Representative genes showed strongly biased expression, including spiderling-biased U3_Pp1a and U5_Pp1e, female-biased U4_Pp1a, and male-biased SMase D_108750 and PaTuSp_108466. These findings reveal sex- and development-biased expression patterns of venom- and silk-related candidate genes in P. astrigera and may provide molecular insights into ecological adaptation and predatory strategies in wandering wolf spiders.

Animals

Reference genome of the Californian trapdoor spider Aptostichus stephencolberti Bond 2008 (Araneae: Mygalomorphae: Euctenizidae).

We present a reference genome assembly for the trapdoor spider Aptostichus stephencolberti. This species, described in 2008, is endemic to the highly fragmented coastal dune habitats of Northern California from Monterey to the San Francisco Bay Area. Trapdoor spiders are ideal taxa for landscape scale genomic studies owing to their extreme site fidelity and limited dispersal capabilities; these same characteristics make them prone to extinction. Genomic studies of species like A. stephencolberti can reveal novel areas of endemism and high conservation value that may not be evident in species with wider ranges and greater dispersal capabilities. As part of the California Conservation Genomics Project, we constructed the A. stephencolberti reference genome from high quality long-read sequences, scaffolded with proximity ligation Omni-C data. The primary assembly comprises 551 scaffolds spanning 3.63 Gbp, a scaffold N50 of 62.2 Mbp and BUSCO completeness of 95.6%. We estimate 52 chromosomes yet find no (TTAGG)n telomer repeats. Expanding the telomeric repeat search finds an ancestral loss of the repeat from all spiders. Automated annotation using the NCBI refseq pipeline and RNAseq data from whole adults finds 14,067 genes with a BUSCO annotation completeness of 95.56%. Repeat annotation identified 77% of the genome to be interspersed repeats. This resource, the first for family Euctenizidae will facilitate future study and resulting conservation actions of A. stephencolberti and other Aptostichus sp. populations associated with the rapidly changing California coastal dune ecosystem.

Aptostichus stephencolberti

Phosphatases in the central nervous system of spiders (Arachnida, Araneae).

The central nervous systems of web-building spiders (Araneidae, Agelenidae) and hunting spiders (Lycosidae, Salticidae) were tested for non-specific and specific phosphatases. Acid phosphatase exhibited weakly to moderately positive reactions in the neuronal cell bodies and in the neuropile fibre mass of all species investigated. Alkaline phosphatase could only be demonstrated in the external and internal neural lamellae of the brain and ventral cord of several specimens of the araneid species investigated. Tests for thiamine pyrophosphatase were negative with both the lead and calcium-cobalt methods. Distinctive positive reactions for adenosine triphosphatase were visible in the nervous system of all the species used, being especially strong in the optic ganglia of the hunting spiders. The demonstration of adenosine triphosphatase was only possible when applying the calcium-cobalt method after Padykula and Herman, while the lead method after Wachstein and Meisel did not produce any staining reaction at all. Controls of the histochemical reaction showed that the enzyme was activated by Ca2+ and inhibited by sulphydryl destroying reagents (e.g. PCMB), but was insensitive to ouabain. It could be probably classified as a mitochondrial proton-translocating adenosine triphosphatase.

Acid Phosphatase

Fine structure of a spider joint receptor and associated synapses.

The fine structure of a joint receptor (R10) in a spider leg (Zygiella x-notata) was examined with light and electron microscopy. The R10 receptor consists of a compact ganglion which is situated near the dorsal joint membrane of the femur/patella joint. Each of the ten sensory cells comprising the ganglion sends one branching dendrite into the hypodermis underlying the joint membrane. All dendritic branches together form a sheet-like meshwork 50 microns wide and 1 microns thick, which is traversed obliquely by hypodermis cells. When the joint is stretched shearing forces are apparently transmitted to the receptive dendritic branches via microtubular bundles inside the hypodermis cells. The soma and dendrites of the sensory cells receive numerous synaptic input from presumably efferent fibres. The fine structure of these synapses is described and compared with other peripheral and central spider synapses. All R10 synapses contain small synaptic vesicles (32 nm diameter), whereas motor endplates possess large vesicles (38 nm). Central synapses have two significantly different vesicle populations which are either of the small or large variety. Since synapses with small vesicles are supposedly inhibitory, receptor cells in spiders might be under efferent control. Such a system is unknown in insects or crustaceans, but may be typical for arachnids.

Animals

Survey of 2144 cases of red-back spider bites: Australia and New Zealand, 1963--1976.

An analysis has been made of 2144 consecutive cases of latrodectism (envenomation by the red-back spider, Latrodectus mactans hasselti) reported to the Commonwealth Serum Laboratories. In the last eight years, notifications have averaged 240 cases per annum. Bites, usually on the extremities (74%), occurred most frequently in the summer months, and in the afternoon or evening. Most victims (79%) were aged between 18 and 50 years and 64.4% of them were males. Males are still often bitten on the genitals or buttocks (9.7% of cases). Local pain, redness and swelling were the most common symptoms, although significant pain was felt at other sites in 39% of the cases. The regional lymph nodes often became swollen and tender within 30 minutes. General effects included nausea, vomiting and sweating, but coma and respiratory failure were very uncommon, probably because of use of specific antivenom. This was administered within 24 hours of the bite in 92% of cases, and within two hours in 70%. Although 11 anaphylactic reactions (0.54%) were reported, no deaths resulted either from the venom or from reactions to the antivenom. Few delayed serum reactions (1.7%) were recorded. The action of the venom is described and the management of red-back spider bite is outlined.

Adolescent

Gene and Genome Duplication in Spiders.

Gene and genome duplications are widely observed across various organisms, including plants, yeasts, and animals. Numerous studies link gene duplications to the emergence of novel phenotypes, supporting the hypothesis that duplication events are advantageous for adaptive evolution. Whole-genome duplications (WGD) are especially prevalent in plants and have also occurred ancestrally in vertebrates. However, large-scale duplication events in other animal groups remain understudied, partly due to limited genomic resources. Arthropods, particularly insects, represent one of the most diverse animal clades in terms of both species and phenotypic diversity. With increasing availability of chromosome-level genomes, large-scale duplications appear to be rare in insects but are more frequent in chelicerates (e.g. spiders, scorpions, and horseshoe crabs). This makes chelicerates an intriguing group for comparing the mechanisms, fates, and evolutionary impacts of large-scale duplications with those seen in plants and vertebrates. In this review, we synthesize and discuss current research on WGD in spiders and discuss different scenarios for genes following gene duplication events (conservation, nonfunctionalization, subfunctionalization, specialization, drift, neofunctionalization) in the context of experimental studies. We hypothesize if there might be common trajectories after duplication and how these could be tested.

Animals

Divergent evolutionary strategies in spider venoms: A comparative proteomic profiling of four sympatric species from Yunnan.

Spider venoms comprise complex cocktails of bioactive molecules evolved for predation and defense, representing a valuable resource for biological research and pharmaceutical discovery. In this study, we performed a systematic analysis of venom gland extracts from four common spider species indigenous to Yunnan, China: Agelena limbata, Hippasa lycosina, Lycosa grahami, and Sinopoda pengi. Using an integrated transcriptomic and proteomic targeted profiling approach, we successfully annotated 141 distinct toxins. Comparative analysis revealed significant interspecific heterogeneity, suggesting distinct evolutionary trajectories and "weapon system economics." Both A. limbata and L. grahami exhibited a "peptide-dominant" profile anchored by neurotoxic peptides and isomerases, optimized for rapid chemical paralysis. In contrast, S. pengi displayed a distinct "protein-dominant" signature enriched with high-molecular-weight enzymes and CAP superfamily proteins, likely functioning to facilitate tissue degradation and toxin diffusion. Occupying an intermediate position, H. lycosina demonstrated a hybrid composition. These findings suggest that although these species share the same geographical range, their venom systems have undergone divergent evolutionary adaptations driven by specific ecological niches and hunting strategies. This study represents the first systematic proteomic characterization of these venom components, providing a valuable reservoir of molecular candidates while highlighting the bioinformatic nuances of analyzing whole-gland homogenates.

Animals

The origin of the muscle fasciculation caused by funnel-web spider venom.

The origin of the fasciculation of skeletal muscle produced by funnel-web spider venom (FSV) has been examined in mouse phrenic nerve hemi-diaphragm preparations, FSV from male spiders at concentrations greater than 10(-6) g/ ml invariably produced muscle fasciculation which could be prevented by d-tubocurarine (14micron), tetrodotoxin (0.3 micron) or by increasing the external magnesium concentration or calcium concentration. Diphenyl hydantoin (3-6 X 10(-5) M) was able to reduce these fasciculations in some experiments. In curarized preparations, multiple end plate potentials (EPPs) in response to single stimuli and bursts of spontaneous EPPs were seen in the presence of FSV (10(-5) g/ml). Extracellular recordings from phrenic nerves in the presence of FSV (10(-5) g/ml) revealed additional components in compound action potentials elicited by single stimuli, and "spontaneous" electrical activity was observed in unstimulated nerves. This spontaneous activity was abolished by raising the divalent cation concentration in the bathing solution. These results suggest that a primary site of action of FSV is the surface membrane of nerve fibres and that muscle fasciculation arises as a consequence of spontaneous action potentials produced by the venom in motor nerves.

Action Potentials

Chromosome-level genome assembly of hawthorn spider mite, Amphitetranychus viennensis (Acari: Tetranychidae).

The hawthorn spider mite, Amphitetranychus viennensis, is a major pest of orchards and ornamentals in the Palaearctic region, with adaptability and acaricide resistance. The lack of high-quality genomic resources limits understanding of its detoxification mechanisms and the development of RNAi-based pest control strategies. In this study, we utilized Illumina, Pacific Biosciences (PacBio), and Hi-C sequencing technologies to assemble a chromosome-level reference genome of A. viennensis. The assembled genome spans 141.96 Mb, with a contig N50 of 1.35 Mb. BUSCO analysis confirmed a high level of completeness, covering 91.6% of annotated genes. The assembly includes 50.97 Mb of repetitive sequences, representing 35.93% of the genome, and annotates 13,968 protein-coding genes. Using Hi-C sequencing, we anchored 47 contigs to three chromosomes, accounting for 97.27% of the estimated nuclear genome and achieving a contig N50 of 45.83 Mb. This high-quality genome assembly provides a valuable foundation for evolutionary and genomic research on spider mites, while also serving as a genetic resource to inform molecular control strategies and support sustainable pest management.

Animals

An analysis of adult social spacing tendencies and related social interactions in a colony of spider monkeys (Ateles geoffroyi) at the San Francisco zoo.

Observations on adult interindividual spacing and social interactions for 13 spider monkeys (Ateles geoffroyi) of an approximately 48 animal captive colony were made from mid-June through August 1972. In this analysis the interanimal distances, general activities, and social interactions per minute observation for each subject were compared for a sample of 7 sexually mature males and 6 sexually mature females. Association tendencies between members of the same sex were significantly high as were exchanges of the taxon-specific 'pectoral sniff and embrane" interactions and grooming. The direction and nature of agonistic interactions was predominantly intersexual and involved frequent attacks by single or multiple males against females. Comparisons with a feral population of spider monkeys (Ateles belzebuth) revealed marked similarities in spatial affinities and patterns of social interaction.

Agonistic Behavior

The unique kidney of the spider monkey (Ateles geoffroyi).

Among nonhuman primates, the renal anatomy of the spider monkey (Ateles geoffroyi) is unique, as it is multipyramidal and multipapillary. Renal function parameters (glomerular filtration rate, renal plasma flow, and concentrating ability) are compared to man and other primates. The kidneys of the spider monkey are similar both anatomically and functionally to man.

Animals

Hepatic amebiasis in spider monkeys.

In the past, investigators have been able to produce hepatic amebiasis in laboratory animals only by direct introduction of parasites into the liver or its vasculature, or by other artificial manipulations. A natural model of human visceral amebiasis has been lacking. We document an extensive outbreak of amebic dystentery which took place in a colony of spider monkeys; severe hepatic abscesses occurred in many animals. The spider monkey is highly susceptible to infection with Entamoeba histolytica and could provide a valuable model for the study of the pathogenesis of invasive amebiasis.

Animals

Vascular spider: a cutaneous manifestation of hyperdynamic blood flow in hepatic cirrhosis.

Arteriography and gas analysis of blood aspirated from a vascular spider in a patient with hepatic cirrhosis were done before excision and histologic examination of the lesion. This study supports earlier work suggesting that a spider angioma is a cutaneous manifestation of a generalized hyperdynamic circulation in hepatic cirrhosis. Moreover, based upon a variety of experimental and clinical observations on the splanchnic circulation in cirrhosis, it is suggested that the portal hypertension blood flow encounters the obstacle posed by the scarred liver ("active congestion").

Arm

Analysis of the venom of the Sydney funnel-web spider, Atrax robustus using gas chromatography mass spectrometry.

Thirteen compounds have been identified using gas chromatography mass spectrometry in the venom of the Sydney funnel-web spider, Atrax robustus. The compounds were identified as their trimethylsilyl or pentafluoropropionate derivatives and were citric acid, lactic acid, phosphoric acid, glycerol, urea, glucose, gamma-aminobutyric acid, glycine, spermidine, spermine, tyramine and octopamine. Female venom contained trace quantities of 5-methyoxytryptamine which was not detected in male venom. Quantitative determination of tyramine and octopamine was achieved using chemical ionization (CH4) gas chromatography mass spectrometry and deuterated internal standards.

Arthropod Venoms

Acetylcholine compartments in mouse diaphragm. Comparison of the effects of black widow spider venom, electrical stimulation, and high concentrations of potassium.

We have studied the effects of 25 mM potassium, electrical stimulation of the phrenic nerve, and crude black widow spider venom on the ultrastructure, electrophysiology, and acetylcholine (ACh) contents of mouse diaphragms. About 65% of the ACh in diaphragms is contained in a depletable store in the nerve terminals. The rest of the ACh is contained in a nondepletable store that may correspond to the store that remains in denervated muscles and includes, in addition, ACh in the intramuscular branches of the phrenic nerve. About 4% of the ACh released from the depletable store at rest is secreted as quanta and may come from the vesicles, while 96% is secreted in a nonquantized form and comes from an extravesicular pool. The size of the extravesicular pool is uncertain: it could be less than 10%, or as great as 50%, of the depletable store. K causes a highly (but perhaps not perfectly) selective increase in the rate of quantal secretion so that quanta account for about 50% of the total ACh released from K-treated diaphragms. K, or electrical stimulation of the phrenic nerve, depletes both the vesicular and extravesicular pools of ACh when hemicholinium no. 3 (HC-3) is present. However, most of the vesicles are retained under these conditions so that the diaphragms are able to increase slightly their rates of release of ACh when K is added. Venom depletes the terminals of their vesicles and abolishes the release of quanta of ACh. It depletes the vesicular pool of ACh (since it depletes the vesicles), but may only partially deplete the extravesicular pool (since it reduces resting release only 10--40%). The rate of release of ACh from the residual extravesicular pool does not increase when 25 mM K is added. Although we cannot exclude the possibility that stimulation may double the rate of release of ACh from the extravesicular pool, our results are compatible with the idea that the ACh released by stimulation comes mainly from the vesicles and that, when synthesis is inhibited by HC-3, ACh may be exchanged between the extravesicular pool and recycled vesicles.

Acetylcholine