A new class of neurotoxin from wasp venom slows inactivation of sodium current

The effects of alpha-pompilidotoxin (alpha-PMTX), a new neurotoxin isolated from the venom of a solitary wasp, were studied on the neuromuscular synapses in lobster walking leg and the rat trigeminal ganglion (TG) neurons. Paired intracellular recordings from the presynaptic axon terminals and the innervating lobster leg muscles revealed that alpha-PMTX induced long bursts of action potentials in the presynaptic axon, which resulted in facilitated excitatory and inhibitory synaptic transmission. The action or alpha-PMTX was distinct from that of other known facilitatory presynaptic toxins, including sea anemone toxins and alpha-scorpion toxins, which modify the fast inactivation of Na+ current. We further characterized the action of alpha-PMTX on Na+ channels by whole-cell recordings from rat trigeminal neurons. We found that alpha-PMTX stowed the Na+ channels inactivation process without changing the peak current-voltage relationship or the activation time course of tetrodotoxin (TTX)-sensitive Na+ currents, and that alpha-PMTX had voltage-dependent effects on the rate of recovery from Na+ current inactivation and deactivating tail currents. The results suggest that alpha-PMTX slows or blocks conformational changes required for fast inactivation of the Na+ channels on the extracellular surface. The simple structure of alpha-PMTX, consisting of 13 amino acids, would be advantageous for understanding the functional architecture of Na+ channel protein.

Identification of bradykinins in solitary wasp venoms

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Isolation and sequence determination of peptides in the venom of the spider wasp (Cyphononyx dorsalis) guided by matrix-assisted laser desorption/ionization time of flight (MALDI-TOF) mass spectrometry

Micro-scale (sub-pmol) isolation and sequence determination of three peptides from the venom of the solitary spider wasp Cyphononyx dorsalis is described. We isolated two novel peptides Cd-125 and Cd-146 and a known peptide Thr(6)-bradykinin from only two venom sacs of solitary spider wasp Cyphononyx dorsalis without bioassay-guided fractionation. but instead guided by MALDI-TOF MS. The MALDI-TOF MS analysis of each fraction showed the purity and molecular weight of the components, which led to the isolation of the peptides virtually without loss of sample amount. The sequences of the novel peptides Cd-125 (Asp-Thr-Ala-Arg-Leu-Lys-Trp-His) and Cd-146 (Ser-Glu-Thr-Gly-Asn-Thr-Val-Thr-Val-Lys-Gly-Phe-Ser-Pro-Leu-Arg) were determined by Edman degradation together with mass spectrometry. and finally corroborated by solid-phase synthesis. The known peptide Thr(6)-bradykinin (Arg-Pro-Pro-Gly-Phe-Thr-Pro-Phe-Arg) was identified by comparison with the synthetic authentic specimen. This is the first example for any kinins to be found in Pompilidae wasp venoms. The procedure reported here can be applicable to studies on many other components of solitary wasp venoms with limited sample availability. (C) 2001 Elsevier B.V. Ltd. All rights reserved.

Novel wasp toxin discriminates between neuronal and cardiac sodium channels

Pompilidotoxins (PMTXs), derived from the venom of solitary wasp has been known to facilitate synaptic transmission in the lobster neuromuscular junction, and a recent further study from rat trigeminal neurons revealed that the toxin slows Na+ channel inactivation without modifying activation process. Here we report that beta -PMTX modifies rat brain type II Na+ channel alpha -subunit (rBII) expressed in human embryonic kidney cells but fails to act on the rat heart alpha -subunit (rH1) at similar concentrations. We constructed a series of chimeric mutants of rBII and rH1 Na+ channels and compared modification of the steady-state Na+ currents by beta -PMTX. We found that a difference in a single amino acid between Glu-1616 in rBII and Gln-1615 in rH1 at the extracellular loop of D4S3-S4 is crucial for the action of beta -PMTX. PMTXs, which are small peptides with 13 amino acids, would be a potential tool for exploring a new functional moiety of Na+ channels.

Sequencing wasp venom peptides by endopeptidase digestion and nested collision-induced dissociation/post-source decay methods

A method incorporating nested collision-induced dissociation/post-source decay (CID/PSD) combined with endopeptidase digestion is described as an approach to determine the sequence of N-terminally modified peptides. The information from immonium and related ions observed in the CID/PSD spectrum was used for the selection of a suitable endopeptidase for the digestion of peptides. Rapid and reliable assignment of peptide sequence was performed by the comparison of CID/PSD spectra of both intact and endopeptidese-digested peptide fragments, since the assignments of the observed fragment ions to either N- or C-terminal ions can thus be carried out unambiguously. This nested CID/PSD method was applied to the sequence determination of two peptides from the solitary wasps Anoplius samariensis and Batozonellus maculifrons (pompilid wasps), which could not be sequenced by the Edman method due to N-terminal modification. Copyright (C) 2002 John Wiley Sons, Ltd.

Winter aggregations, colony cycle, and seasonal phenotypic change in the paper wasp Polistes versicolor in subtropical Brazil

Social wasps from temperate zones have clear annual colony cycles, and the young queens hibernate during winter. In the subtropics, the only previously reported evidence for the existence of hibernation is the facultative winter aggregations of females during harsh climate conditions. As in temperate-zone species analyzed so far, we show in this study that in the paper wasp, Polistes versicolor, a subtropical species, body size increases as an unfavorable season approaches. Our morphological studies indicate that larger females come from winter aggregations-that is, they are new queens. Multivariate analyses indicate that size is the only variable analyzed that shows a relationship to the differences. Given the absence of a harsh climate, we suggest that the occurrence of winter aggregations in tropical P. versicolor functions to allow some females to wait for better environmental conditions to start a new nest, rather than all being obliged to start a new nest as soon as they emerge.

Eumenitin, a novel antimicrobial peptide from the venom of the solitary eumenine wasp Eumenes rubronotatus

A novel antimicrobial peptide, eumenitin, was isolated from the venom of the solitary eumenine wasp Eumenes rubronotatus. The sequence of eumenitin, Leu-Asn-Leu-Lys-Gly-Ile-Phe-Lys-Lys-Val-Ala-Ser-Leu-Leu-Thr, was mostly analyzed by mass spectrometry together with Edman degradation, and corroborated by solid-phase synthesis. This peptide has characteristic features of cationic linear a-helical antimicrobial peptides, and therefore, can be predicted to adopt an amphipathic a-helix secondary structure. In fact, the CD spectra of eumenitin in the presence of TFE or SDS showed a high content of alpha-helical conformation. Eumenitin exhibited inhibitory activity against both Gram-positive and Gram-negative bacteria, and moderately stimulated degranulation from the rat peritoneal mast cells and the RBL-2H3 cells, but showed no hemolytic activity against human erythrocytes. This antimicrobial peptide in the eumenine wasp venom may play a role in preventing potential infection by microorganisms during prey consumption by their larvae. (c) 2006 Elsevier B.V. All rights reserved.

Decoralin, a novel linear cationic alpha-helical peptide from the venom of the solitary eumenine wasp Oreumenes decoratus

A novel peptide, decoralin, was isolated from the venom of the solitary eumenine wasp Oreumenes decoratus. its sequence, Ser-Leu-Leu-Ser-Leu-Ile-Arg-Lys-Leu-Ile-Thr, was determined by Edman degradation and corroborated by solid-phase synthesis. This sequence has the characteristic features of linear cationic a-helical peptides; rich in hydrophobic and basic amino acids with no disulfide bond, and accordingly, it can be predicted to adopt an amphipathic a-helix secondary structure. In fact, the CD spectra of decoralin in the presence of TFE or SDS showed a high a-helical conformation content. In a biological evaluation, decoralin exhibited a significant broad-spectrum antimicrobial activity, and moderate mast cell degranulation and leishmanicidal activities, but showed virtually no hemolytic activity. A synthetic analog with C-terminal amidation showed a much more potent activity in all the biological assays. (c) 2007 Elsevier B.V. All rights reserved.

Morphological caste differences and non-sterility of workers in Brachygastra augusti (Hymenoptera, Vespidae, Epiponini), a neotropical swarm-founding wasp

Morphological caste differences and ovary conditions were analyzed in four colonies of Brachygastra augusti collected in different stages of the colony cycle. Differences between castes are distinct suggesting pre-imaginal determination. Many intermediates (non-inseminated ovary-developed females) were found. Because these intermediate females were morphologically similar to workers, it is suggested that they are young ovary-developed workers. Because there is a more evident overlap between queens and workers in the colony in pre-emergence of workers stage, and in three colonies in latter stages, especially in worker-production colonies, it is suggested that smaller queens are probably less viable than larger queens as observed in previously studied epiponines.

Anoplin, a novel antimicrobial peptide from the venom of the solitary wasp Anoplius samariensis

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Polybitoxins: A group of phospholipases A2 from the venom of the neotropical social wasp paulistinha (Polybia paulista)

The neotropical wasp Polybia paulista is very aggressive and endemic in south-east Brazil, where it frequently causes stinging accidents. By using gel filtration on Sephadex G-200, followed by ion-exchange chromatography on DEAE-Cellulose under a pH gradient, a group of four toxins (designated as polybitoxins-I, II, lII and IV) presenting phospholipase A2 (PLA2) activities was purified. These toxins are dimeric with mol. wts ranging from 115,000 to 132,000 and formed by different subunits. The four toxins contain very high sugar contents attached to their molecules (22-43% w/w) and presented different values of pH optimum from 7.8 to 9.0; when dissociated, only residual catalytic activities were maintained. The catalytic activities of polybitoxins (from 18 to 771 μmoles/mg per minute) are lower than that of PLA2 from Apis mellifera venom and hornetin from Vespa basalis. The polybitoxins presented a non-linear steady-state kinetic behavior for the hydrolysis of phosphatidylcholine at pH 7.9, compatible with the negative co- operativity phenomena. All of the polybitoxins were very potent direct hemolysins, especially the polybitoxins-III and IV, which are as potent as the lethal toxin from V. basalis and hornetin from Vespa flavitarsus, respectively; polybitoxin-IV presented hemolytic action 20 times higher than that of PLA2 from A. mellifera, 17 times higher than that of neutral PLA2 from Naja nigricolis and about 37 times higher than that of cardiotoxin from Naja naja atra venom.

Social organization of the eusocial wasp Mischocyttarus cerberus styx (Hymenoptera, Vespidae)

The present study was carried out under field conditions in Rio Claro, Southeastern Brazil. The following average life-spans of adults of Mischocyttarus cerberus styx were observed: queens 191.1 days, workers 14.0 days, and males 10.7 days. The short longevity of workers is attributed to the early foraging activity which was performed since the first week of adult age. The life table of the workers revealed high mortality rates during the first three weeks of life. Twenty-four behavioral categories were reported for these wasps and a comparative behavioral catalogue showed that the solitary foundresses (pre-emergence stage) are significantly more active than the queens (post-emergence stage) because they have to perform all the tasks for the nest maintenance. On the other hand, the queens were highly active even in the presence of the workers. The small number of individuals on the nest was the probable cause of the high activity of the queens in the post-emergence colonies. There was no clear division of tasks among workers according to their age (in weeks), so that most of the behavior categories were potentially performed by any worker of any age. Dead queens were replaced by their daughters.

Dominance hierarchy in different stages of development in colonies of the primitively eusocial wasp Mischocyttarus cerberus styx (Hymenoptera, Vespidae)

In primitively eusocial insect societies, reproductive division of labor is established by dominance-submission interactions which determine a linear dominance hierarchy. As previously observed for other species, in Mischocyttarus cerberus styx (Hymenoptera, Vespidae), the dominant female is the main egg layer. Most of her attacks were to the females directly beneath in the rank establishing a hierarchy. During the pre-emergence stage, the hierarchy was already defined and in the post-emergence stage, pre-male substage, the frequency of dominance interactions were strong but the hierarchy was still maintained by the first-ranked female. In the decline stage there were many neutral individuals which could be the reproductives of a new colony.

Patterns of honey storage in nests of the neotropical paper wasp Polistes simillimus Zikán, 1951 (Hymenoptera, Vespidae)

This study describes, for the first time, the occurrence of two patterns of honey storage in nests of the paper wasp Polistes simillimus. During the period of January of 1997 to September of 2000 we visited 262 colonies of P. simillimus, found at several places in States of São Paulo and Minas Gerais, in the southeast region of Brazil. Only in 2.67% of the colonies of P. simillimus searched presented the behavior of honey storage. Two of these colonies were in the pre-emergency phase, two in the post-emergency and three in decline. The results suggest that the honey storage can be correlated with the number of present individuals in the colony, because the colonies in decline, with larger number of individuals, presented many more cells occupied with honey than the nests in pre and post-emergency. There was not a pattern for the distribution of the cells with honey in the nests in decline. The honey storage in the colonies in pre and post-emergency of P. simillimus, suggests a strategy to increase success of foraging activity in the function of parental care. Already for the colonies in decline, this strategy seems to be associated with the individuals' survival in the aggregation, since the wasps are awaiting the passage of the unfavorable climatic conditions for the dispersion and foundation of new colonies.

Influence of the colony cycle on physiological and morphological caste variation in the perennial neotropical swarm-founding social wasp, Protonectarina sylveirae (Hymenoptera, Vespidae, Epiponini)

This paper describes the influence of the colony cycle on caste differences in Protonectarina sylveirae. Despite invariably keeping the main aspects of the conspicuous dimorphism between the castes, it is shown that: 1- queen-worker differences varied according to the progress of the colony cycle, 2- queens in pre-emergence colonies were smaller than those in male-producing ones, 3- workers were larger in pre-emergence than in other colony stages, 4- at particular steps of the colony cycle, non-inseminated ovary-developed queens and workers were detected. The size of the spermatheca differed between inseminated and non-inseminated queens with the highest values appearing in those bearing the most developed kind of ovary development. Differences between inseminated and non-inseminated queens were found in tergites III and IV, ovarian development and amount of fatty tissues. Because tergite size relates to gaster size, this character may be an important stimulus for selection of larger queens in the course of the colony cycle.