Toxicity during early development of the mouse nervous system of a scorpion neurotoxin active on sodium channels

The lethal effects of scorpion envenomation is due to neurotoxins active on voltage-sensitive sodium channels. Dysfunctions of the peripheral and central nervous systems with neurological manifestations are commonly observed after scorpion stings, specially in young children. Since the neurotoxicity of venom fraction is greatly higher by intracerebroventricular than by subcutaneous injections, a direct effect of venom on CNS cannot be excluded specially in infants where the blood-brain barrier is not fully functional. We investigated the activity of a neurotoxin from the scorpion Androctonus australis hector (AahII) in newborn mice at 3, 7 and 14 days after birth and in adults. Young mice (P3, P7) were more sensitive to AahII injected subcutaneously than were adults, but were less sensitive to intracerebroventricular injection. The affinity of AahII for its receptor site on brain synaptosomes from P3 and P7 mice was slightly higher and the density of the binding sites was half that of adult mice. After subcutaneous injection of [125I]-AahII it was also observed that a small amount of radioactivity was found in brains of neonate mice but not in that of adults. This amount is however extremely lower than the value of the LD50 determined by intracerebroventricular injection. Results are consistent with a peripheral action of AahII and show that its toxic activity changes during the mouse nervous system development.     

Two New Species of Coronigoniella Young from Brazil with Taxonomic and Distributional Notes on the Genus (Hemiptera: Cicadellidae: Cicadellinae)

Two new species of Coronigoniella Young are described and illustrated: C. osborni, sp. n., from SE. and S. Brazil (Espírito Santo, Minas Gerais, Rio de Janeiro, and Paraná states), and C. formosa, sp. n., from central W. and SE. Brazil (Goiás and Minas Gerais states, and the Distrito Federal). The presence of a sclerotized plate from the abdominal sternum VIII is reported in females of Coronigoniella for the first time. Taxonomic and morphological notes are given for C. spinosa (Osborn) based on specimens from its type-locality in Mato Grosso State. Tettigonia ruficaput (Walker), a species from Colombia previously considered of uncertain taxonomic position, is transferred to Coronigoniella. Possible host plants of C. osborni, sp. n. are Coleus sp. (Lamiaceae), Symphytum sp. (Boraginaceae), and Vernonia sp. (Asteraceae). The genus Coronigoniella is newly recorded from Alagoas, Bahia, Espírito Santo, Goiás, Minas Gerais, Paraná, and São Paulo states, as well as from the Distrito Federal, Brazil. A map showing the known geographic distribution of the two new species is presented. Resumo Duas novas espécies de Coronigoniella Young são descritas e ilustradas: C. osborni, sp. n., do sudeste e sul do Brasil (estados do Espírito Santo, Minas Gerais, Rio de Janeiro e Paraná), e C. formosa, sp. n., do centro-oeste e sudeste do Brasil (estados de Goiás e Minas Gerais, assim como o Distrito Federal). A presença de uma placa esclerosada do esterno abdominal VIII é registrada em fêmeas de Coronigoniella pela primeira vez. Notas taxonômicas e morfológicas sobre C. spinosa (Osborn) são fornecidas, com base em espécimes provenientes da sua localidade-tipo no Estado do Mato Grosso. Tettigonia ruficaput (Walker), uma espécie da Colômbia anteriormente considerada como de posição taxonômica incerta, é transferida para Coronigoniella. Coleus sp. (Lamiaceae), Symphytum sp. (Boraginaceae) e Vernonia sp. (Asteraceae) são possíveis plantas hospedeiras de C. osborni, sp. n. O gênero Coronigoniella é pela primeira vez registrado nos estados de Alagoas, Bahia, Espírito Santo, Goiás, Minas Gerais, Paraná e São Paulo, assim como no Distrito Federal, Brasil. Um mapa mostrando a distribuição geográfica conhecida das duas novas espécies é apresentado.     

Was high grouse hag in early 20th century Norway due to a program for extermination of small game predators?

A Norwegian program for extermination of (small game) predators (NPEP) was run during 1900‐ 1914, This initiative is believed to have caused larger small game stocks and more regional synchrony in rodents. To investigate the effectiveness of the NPEP time series of predators bountied (1885–1914), rodent dynamics (1885–1914), ptarmigan hunting index (1885–1914 and 1900/1‐1914), and willow ptarmigan and berry export statistics (1885–1914) were analyzed for three different regions: south, east and central Norway. In south and east Norway there were higher ptarmigan export in the period 1907–1914 than the years before, but not in central Norway. There was not bountied higher number of red fox. eagle or goshawk in any of the three regions when comparing years a) before and after 1900. and b) when comparing the periods 1900–1906 and 1907–1914. This suggests that predator removal was not the cause of increased ptarmigan export. The ptarmigan hunting index and rodent index for south and east Norway were correlated in the period 1885 1914, while the ptarmigan export from east Norway was correlated with berry export from the year before. However, for central Norway the rodent index was not correlated with the hunting index. There were cross correlation between berry and ptarmigan export with lag from minus one to nine years for south and east Norway, The NAO (North Atlantic Oscillation) ‐ an indication of the winter weather variation ‐ had higher values during 1900–1914 than 1885–1899, indicating moister and warmer winters in the last period. This analysis indicates that NPEP generally did not increase predator removal. The results suggest, however, that it was a series of years with high rodent density, that caused the increase in ptarmigan populations In south and cast Norway, which, in turn, may have been caused by favourable weather conditions leading to among others good berry crops. Conclusions based on old data series must, however, be drawn with caution.     

A new feather mite species of the genus <Emphasis Type="Italic">Picalgoides</Emphasis> Černý, 1974 (Astigmata: Psoroptoididae) from a passerine host in Costa Rica

A new feather mite species, Picalgoides giganteus n. sp. (Psoroptoididae: Pandalurinae), is described from the tawny-throated leaftosser Sclerurus mexicanus Sclater (Passeriformes: Furnariidae) in Costa Rica. Among the 10 species of Picalgoides Černy, 1974, including the new one, this is the third recorded from a passerine host; the remaining seven nominal species are associated with hosts of the order Piciformes. Brief data on the host-parasite associations of Picalgoides spp. are provided. Megninia megalixus Trouessart, 1885 from the short-tailed green magpie Cissa thalassina (Temminck) is transferred to Picalgoides as P. megalixus (Trouessart, 1885) n. comb.     

The Lower Devonian scorpionWaeringoscorpio and the respiratory nature of its filamentous structures, with the description of a new species from the Westerwald area, Germany

The fossil scorpionWaeringoscorpio hefteri Størmer, 1970 (Arachnida: Scorpiones) from the Lower Devonian of the Rhenish Massif of Germany is redescribed based on both the original type and newly collected material. A second, more tuberculate species from Siegenian strata near Bürdenbach in the Westerwald (also part of the Rhenish Massif) is described asW. westerwaldensis n. sp. Details of the coxo-sternal region — including the lack of an oral tube — and the number of ventral mesosomal plates are discussed.Waeringoscorpio Størmer, 1970 is best known for its possession of externally-projecting ‘gills’. Our new material reveals that these are indeed pair-wise bundles of rigid, branching filaments which originate laterally, quite possibly from those segments of the mesosoma associated with the book lungs in extant scorpions. Their gross morphology is most consistent with a respiratory organ adapted for use in water. Indeed their closest modern analogues are the tracheal gills of secondarily aquatic insects. We suggest that the morphology and likely palaeoenvironment ofWaeringoscorpio could indicate an aquatic animal, but we draw attention to the uniqueness of its gill-structures, which may not be part of the scorpion ground-pattern. Thus,Waeringoscorpio was perhaps a secondarily aquatic scorpion adapted for benthic life in oxygen-stressed, freshwater-brackish environments.     

Redescription and palaeobiology of Palaeoscorpius devonicus Lehmann, 1944 from the Lower Devonian Hunsrück Slate of Germany

Abstract: Palaeoscorpius devonicus Lehmann, 1944 is known from only a single specimen, found in the Eschenbach Pit near Bundenbach in the Lower Devonian Hunsrück Slate of Germany. It is a key fossil, having been interpreted both as the most basal member of the Scorpiones and as one of the order’s most likely candidates for an aquatic mode of life. Prepared both ventrally and dorsally, some aspects of its morphology remain problematic. Here, with the aid of new techniques, including computed tomography, we present a re‐investigation of this scorpion’s anatomy and a new reconstruction, with a particular focus on the species’ original habitat. On the basis of the environmental interpretation of the Hunsrück Slate and the completeness of the specimen, previous authors concluded that P. devonicus was marine, but none offered convincing morphological evidence. Recent studies of the deposit’s environment suggest that the Hunsrück Sea was part of an intrashelf basin, relatively close to the coastline, and fossils of land plants show that terrestrial wash‐in occasionally occurred. Our revised interpretation of the fossil’s morphology demonstrates that the scorpion was most probably terrestrial. Internal mesosomal organs are interpreted as book lungs, but other terrestrial adaptations are lacking. The absence of both coxapophyses and gnathobases makes determining the scorpion’s feeding mechanism difficult. Interpreting the scorpion’s character states within a phylogenetic framework, especially the possible presence of book lungs, implies either that the plesiomorphic position of P. devonicus is no longer supported or that the development of book lungs had already taken place early in the scorpion lineage.     

Gene responsible for protecting Escherichia coli from sodium dodecyl sulfate and toluidine blue plus light.

Escherichia coli cells were killed by visible light irradiation in the presence of the photosensitizing dye, toluidine blue. Two uvrB mutant strains of E. coli K-12 (AB1885 and N3-1) were much more sensitive than the isogenic uvrA and uvrC strains to treatment with toluidine blue plus light, suggesting that the uvrB+ gene product was involved in repair of DNA damage induced by the treatment. The uvrB+ gene cloned in a high- or low-copy-number plasmid was transformed into the uvrB strain (AB1885). Although all the transformants showed the same resistance as its wild-type strain (AB1157) to UV irradiation, they were as sensitive as AB1885 was to treatment with toluidine blue plus light. The two uvrB strains were more sensitive to sodium dodecyl sulfate than the other strains, suggesting that these strains had a defect in the cell surface. A sodium dodecyl sulfate-resistant revertant obtained from AB1885 was more resistant than AB1885 was to treatment with toluidine blue plus light. The two uvrB strains (AB1885 and N3-1) appear to have a defective gene (tentatively called dvl) different from uvrB. Its map position was around 7 min on the E. coli map.     

Genotypic differences in phosphorus efficiency of wheat

In an attempt to evaluate whether breeding and selection for high yielding capacity did change the P requirements of modern wheat cultivars, the response of two wheat cultivars to different levels of P supply was investigated. A traditional cultivar ("Peragis") and a modern cultivar ("Cosir") were cultivated in a C-loess low in available P and high in CaCO₃ in 120 cm high PVC pots. Shoot and root growth at different developmental stages was compared. The grain yield of the modern cultivar Cosir was higher at limiting and non-limiting P supply and, therefore, this cultivar can be considered as more P-efficient than the traditional cultivar. From the results it can be concluded that the main factors contributing to the higher P efficiency of the modern cultivar are (i) efficient use of assimilates for root growth characteristics which enhance P acquisition: smaller root diameter, and longer root hairs, (ii) efficient remobilization of P from vegetative plant organs to the grains, and (iii) lower P requirement for grain yield formation because of lower ear number per plant but higher grain number per ear.     

A spider toxin that induces a typical effect of scorpion alpha-toxins but competes with beta-toxins on binding to insect sodium channels

Delta-palutoxins from the spider Paracoelotes luctuosus (Araneae: Amaurobiidae) are 36-37 residue long peptides that show preference for insect sodium channels (NaChs) and modulate their function. Although they slow NaCh inactivation in a fashion similar to that of receptor site 3 modifiers, such as scorpion alpha-toxins, they actually bind with high affinity to the topologically distinct receptor site 4 of scorpion beta-toxins. To resolve this riddle, we scanned by Ala mutagenesis the surface of delta-PaluIT2, a delta-palutoxin variant with the highest affinity for insect NaChs, and compared it to the bioactive surface of a scorpion beta-toxin. We found three regions on the surface of delta-PaluIT2 important for activity: the first consists of Tyr-22 and Tyr-30 (aromatic), Ser-24 and Met-28 (polar), and Arg-8, Arg-26, Arg-32, and Arg-34 (basic) residues; the second is made of Trp-12; and the third is made of Asp-19, whose substitution by Ala uncoupled the binding from toxicity to lepidopteran larvae. Although spider delta-palutoxins and scorpion beta-toxins have developed from different ancestors, they show some commonality in their bioactive surfaces, which may explain their ability to compete for an identical receptor (site 4) on voltage-gated NaChs. Yet, their different mode of channel modulation provides a novel perspective about the structural relatedness of receptor sites 3 and 4, which until now have been considered topologically distinct.     

Common features in the functional surface of scorpion beta-toxins and elements that confer specificity for insect and mammalian voltage-gated sodium channels

Scorpion beta-toxins that affect the activation of mammalian voltage-gated sodium channels (Navs) have been studied extensively, but little is known about their functional surface and mode of interaction with the channel receptor. To enable a molecular approach to this question, we have established a successful expression system for the anti-mammalian scorpion beta-toxin, Css4, whose effects on rat brain Navs have been well characterized. A recombinant toxin, His-Css4, was obtained when fused to a His tag and a thrombin cleavage site and had similar binding affinity for and effect on Na currents of rat brain sodium channels as those of the native toxin isolated from the scorpion venom. Molecular dissection of His-Css4 elucidated a functional surface of 1245 A2 composed of the following: 1) a cluster of residues associated with the alpha-helix, which includes a putative "hot spot" (this cluster is conserved among scorpion beta-toxins and contains their "pharmacophore"); 2) a hydrophobic cluster associated mainly with the beta2 and beta3 strands, which is likely to confer the specificity for mammalian Navs; 3) a single bioactive residue (Trp-58) in the C-tail; and 4) a negatively charged residue (Glu-15) involved in voltage sensor trapping as inferred from our ability to uncouple toxin binding from activity upon its substitution. This study expands our understanding about the mode of action of scorpion beta-toxins and illuminates differences in the functional surfaces that may dictate their specificities for mammalian versus insect sodium channels.     

Contractile activation in scorpion striated muscle fibers. Dependence on voltage and external calcium

Excitation-contraction coupling was characterized in scorpion striated muscle fibers using standard microelectrode techniques as employed in studies on vertebrate skeletal muscle. The action potential of scorpion muscle consists of two phases of regenerative activity. A relatively fast, overshooting initial spike is followed by a prolonged after- discharge of smaller, repetitive spikes. This after-discharge is accompanied by a twitch that relaxes promptly upon repolarization. Twitches fail in Na-free, tetrodotoxin (TTX)-containing, or Ca-free media. However, caffeine causes contractures in muscles paralyzed by Na- and Ca-free solutions. Experiments on muscle fibers voltage-clamped at a point with two microelectrodes in Na-free or TTX-containing media indicate that: (a) the strength-duration relation for threshold contractions has a shape similar to that in frog muscle, but mean values are displaced approximately 20 mV in the positive direction; (b) tetracaine exerts a parallel effect on strength-duration curves from scorpion and frog; (c) contractile activation in scorpion is abolished in Ca-free media; and (d) the contractile threshold is highly correlated with the occurrence of inward Ca current for pulses of all durations. Thus, the voltage dependence of contractile activation in scorpion and frog muscle is similar. However, the preparations differ in their dependence on extracellular Ca for contraction. These results are discussed in relation to possible mechanisms coupling tubular depolarization to Ca release from the sarcoplasmic reticulum in vertebrate and invertebrate skeletal muscle.     

蝎毒素基因分子生物学研究进展

介绍了克隆蝎毒素cDNA和基因组基因的策略以及蝎毒素cDNA的结构和毒素蛋白前体的翻译后加工过程,同时综述了蝎毒素基因组基因的组织结构及其mRNA前体的加工以及重组蝎毒素基因表达的研究进展.     

Binding of scorpion toxin to receptor sites associated with sodium channels in frog muscle. Correlation of voltage-dependent binding with activation.

Purified scorpion toxin (Leiurus quinquestriatus) slows inactivation of sodium channels in frog muscle at concentrations in the range of 17-170 nM. Mono[125I]iodo scorpion toxin binds to a single class of sites in frog sartorius muscle with a dissociation constant of 14 nM and a binding capacity of 13 fmol/mg wet weight. Specific binding is inhibited more than 90% by 3 microM sea anemone toxin II and by depolarization with 165 mM K+. Half-maximal inhibition of binding is observed on depolarization to -41 mV. The voltage dependence of scorpion toxin binding is correlated with the voltage dependence of activation of sodium channels. Removal of calcium from the bathing medium shifts both activation and inhibition of scorpion toxin binding to more negative membrane potentials. The results are considered in terms of the hypothesis that activation of sodium channels causes a conformational change in the scorpion toxin receptor site resulting in reduced affinity for scorpion toxin.     

Solution structure of kurtoxin: a gating modifier selective for Cav3 voltage-gated Ca(2+) channels

Kurtoxin is a 63-amino acid polypeptide isolated from the venom of the South African scorpion Parabuthus transvaalicus. It is the first and only peptide ligand known to interact with Cav3 (T-type) voltage-gated Ca(2+) channels with high affinity and to modify the voltage-dependent gating of these channels. Here we describe the nuclear magnetic resonance (NMR) solution structure of kurtoxin determined using two- and three-dimensional NMR spectroscopy with dynamical simulated annealing calculations. The molecular structure of the toxin was highly similar to those of scorpion α-toxins and contained an α-helix, three β-strands, and several turns stabilized by four disulfide bonds. This so-called "cysteine-stabilized α-helix and β-sheet (CSαβ)" motif is found in a number of functionally varied small proteins. A detailed comparison of the backbone structure of kurtoxin with those of the scorpion α-toxins revealed that three regions [first long loop (Asp(8)-Ile(15)), β-hairpin loop (Gly(39)-Leu(42)), and C-terminal segment (Arg(57)-Ala(63))] in kurtoxin significantly differ from the corresponding regions in scorpion α-toxins, suggesting that these regions may be important for interacting with Cav3 (T-type) Ca(2+) channels. In addition, the surface profile of kurtoxin shows a larger and more focused electropositive patch along with a larger hydrophobic surface compared to those seen on scorpion α-toxins. These distinct surface properties of kurtoxin could explain its binding to Cav3 (T-type) voltage-gated Ca(2+) channels.     

ARVC-related mutations in divergent region 3 alter functional properties of the cardiac ryanodine receptor

Two single-nucleotide polymorphisms in the type 2 ryanodine receptor (RyR2) leading to the nonsynonymous amino acid replacements G1885E and G1886S are associated with arrhythmogenic right ventricular cardiomyopathy in patients who are carrying both of the corresponding RyR2 alleles. The functional properties of HEK293 cell lines isogenically expressing RyR2 mutants associated with arrhythmogenic right ventricular cardiomyopathy, RyR2-G1885E, RyR2-G1886S, RyR2-G1886D (mimicking a constitutively phosphorylated Ser¹⁸⁸⁶), and the double mutant RyR2-G1885E/G1886S were investigated by analyzing the intracellular Ca²⁺ release activity resulting from store-overload-induced calcium release. The substitution of serine for Gly¹⁸⁸⁶ caused a significant increase in the cellular Ca²⁺ oscillation activity compared with RyR2 wild-type-expressing HEK293 cells. It was even more pronounced if glycine 1885 or 1886 was replaced by the acidic amino acids glutamate (G1885E) or aspartate (G1886D). Surprisingly, when both substitutions were introduced in the same RyR2 subunit (RyR2-G1885E/G1886S), the store-overload-induced calcium release activity was nearly completely abolished, although the Ca²⁺ loading of the intracellular stores was markedly enhanced, and the channel still displayed substantial Ca²⁺ release on stimulation by 5 mM caffeine. These results suggest that the adjacent glycines 1885 and 1886, located in the divergent region 3, are critical for the function and regulation of RyR2.     

Molecular cloning and nucleotide sequence analysis of encoded anti-insect toxin BotIT2 from the scorpion Buthus occitanus tunetanus venom

Numerous toxins from scorpion venoms are much more toxic to insects than to other animal classes, and possess high affinity to Na+ channels. Many of them active on insects were purified from the venom of Buthus occitanus tunetanus. Using amino acid sequences of BotIT2 and RACE-PCR amplification (Rapid amplification of cDNA ends) technique, we isolated, identified and sequenced the nucleotide sequence from the venom glands of the scorpion Buthus occitanus tunetanus. The cDNA encodes a precursor of an insect toxin of 60 amino acid residues. The deduced nucleotide sequence toxin was identical to the determined amino acid sequence of BotIT2. BotIT2 is more similar to the excitatory toxins in its mode of action and to the depressant toxins in its primary structure.     

Anti-tumoral effect of scorpion peptides: Emerging new cellular targets and signaling pathways

Scorpion toxins have been the subject of many studies exploring their pharmacological potential. The high affinity and the overall selectivity to various types of ionic channels endowed scorpion toxins with a potential therapeutic effect against many channelopathies. These are diseases in which ionic channels play an important role in their development. Cancer is considered as a channelopathy since overexpression of some ionic channels was highlighted in many tumor cells and was linked to the pathology progression.Interestingly, an increasing number of studies have shown that scorpion venoms and toxins can decrease cancer growth in vitro and in vivo. Furthermore through their ability to penetrate the cell plasma membrane, certain scorpion toxins are able to enhance the efficiency of some clinical chemotherapies. These observations back-up the applicability of scorpion toxins as potential cancer therapeutics.In this review, we focused on the anti-cancer activity of scorpion toxins and their effect on the multiple hallmarks of cancer. We also shed light on effectors and receptors involved in signaling pathways in response to scorpion toxins effect. Until now, the anticancer mechanisms described for scorpion peptides consist on targeting ion channels to (i) inhibit cell proliferation and metastasis; and (ii) induce cell cycle arrest and/or apoptosis through membrane depolarization leading to hemostasis deregulation and caspase activation. Putative targets such as metalloproteinases, integrins and/or growth factor receptors, beside ion channels, have been unveiled to be affected by scorpion peptides.