RESUMO
A useful approach to deepen our knowledge about the origin and evolution of venom systems in Reptilia has been exploring the vast biodiversity of this clade of vertebrates in search of orally produced proteins with toxic actions, as well as their corresponding delivery systems. The occurrence of toxins in anguimorph lizards has been demonstrated experimentally or inferred from reports of the toxic effects of the oral secretions of taxa within the Varanidae and Helodermatidae families. In the present study, we have focused on two alligator lizards of the Anguidae family, the Mexican alligator lizard, Abronia graminea, and the red-lipped arboreal alligator lizard, A. lythrochila. In addition, the fine morphology of teeth of the latter species is described. The presence of a conserved set of proteins, including B-type natriuretic peptides, cysteine-rich secretory proteins, group III phospholipase A2, and kallikrein, in submandibular gland extracts was demonstrated for both Abronia species. These proteins belong to toxin families found in oral gland secretions of venomous reptile species. This finding, along with previous demonstration of toxin-producing taxa in both paleo- and neoanguimorpha clades, provides further support for the existence of a handful of conserved toxin families in oral secretions across the 100+ million years of Anguimorpha cladogenesis.
RESUMO
Some butterflies and moths sequester and retain noxious phytochemicals for defence against predators. In the present study, three moth species, the garden tiger moth, Arctia caja, the death hawk moth, Acherontia atropos, and the oleander hawk moth, Daphnis nerii, were tested whether they sequester alkaloids from their host plants. Whereas A. caja consistently sequestered atropine from Atropa belladonna, also when atropine sulfate was added to the alkaloid-free diet of the larvae, A. atropos and D. nerii were found to be unable to sequester alkaloids, neither atropine nor eburnamenine from Vinca major, respectively. Instead of acquiring toxicity as chemical defence, nocturnal lifestyle and cryptic attitudes may improve their chances of survival.
Assuntos
Alcaloides , Borboletas , Mariposas , Animais , Larva , Derivados da AtropinaRESUMO
Based on its phylogenetic relationship to monitor lizards (Varanidae), Gila monsters (Heloderma spp.), and the earless monitor Lanthanotus borneesis, the Chinese crocodile lizard, Shinisaurus crocodilurus, has been assigned to the Toxicofera clade, which comprises venomous reptiles. However, no data about composition and biological activities of its oral secretion have been reported. In the present study, a proteomic analysis of the mandibular gland of S. crocodilurus and, for comparison, of the herbivorous Solomon Island skink Corucia zebrata, was performed. Scanning electron microscopy (SEM) of the teeth from S. crocodilurus revealed a sharp ridge on the anterior surface, but no grooves, whereas those of C. zebrata possess a flattened crown with a pointed cusp. Proteomic analysis of their gland extracts provided no evidence of venom-derived peptides or proteins, strongly supporting the non-venomous character of these lizards. Data are available via ProteomeXchange with identifier PXD039424.
Assuntos
Jacarés e Crocodilos , Lagartos , Animais , Jacarés e Crocodilos/metabolismo , Lagartos/metabolismo , Filogenia , Proteômica , Peçonhas/químicaRESUMO
Over the past decades, envenomation by caterpillars of Automeris spp. became an increasing health problem in Latin America. Accidental contact with the stinging spines of these caterpillars cause acute local pain, itching, inflammation and skin rashes that persists for days. Even when the cause is obvious, the exact molecular mechanisms responsible for the observed symptoms are yet to be elucidated. Here, we describe for the first time, an active compound in the venom and the study of the bioactivity of the venom extracted from the spines of the caterpillar Automeris zaruma. Electrophysiological screening of a library of membrane proteins important for pain and itch enabled us to investigate and reveal the mode of action of the venom of A. zaruma. Further mass spectrometric analysis (Q-TOF-MS) made it possible to establish a link between the bioactivity and the components found in the venom. We show that the spine extract of A. zaruma contains histamine that potently activates the four types of the human histamine receptors (H1R, H2R, H3R and H4R) with a selectivity preference towards H3R and H4R. Furthermore, a modulation of the target MRGPRX2 was found. Together, these findings are the first to explain the symptomology of A. zaruma envenomation, enabling us a better understanding of caterpillar envenomation and predict that the hurdle of the scarce efficacy of the currently used antihistaminic drugs can be overcome by including H3R and H4R blockers in the clinical used medication. Such an approach might be used for other caterpillar envenomation in the world and represent a significant improvement for the well-being of the patient.
Assuntos
Histamina , Manduca , Receptores Histamínicos , Peçonhas , Animais , Histamina/metabolismo , Humanos , Lepidópteros , Proteínas do Tecido Nervoso/genética , Proteínas do Tecido Nervoso/metabolismo , Dor/etiologia , Prurido/etiologia , Receptores Acoplados a Proteínas G/genética , Receptores Acoplados a Proteínas G/metabolismo , Receptores Histamínicos/genética , Receptores Histamínicos/metabolismo , Receptores Histamínicos H4/genética , Receptores Histamínicos H4/metabolismo , Receptores de Neuropeptídeos/genética , Receptores de Neuropeptídeos/metabolismo , Peçonhas/efeitos adversos , Peçonhas/química , Peçonhas/metabolismoRESUMO
Rough-skinned newts, Taricha granulosa, which contain tetrodotoxin (TTX), a potent neurotoxin, are widely distributed along the west-coast of North America up to British Columbia (BC), Canada, and Southeast Alaska. Their genetic population structure using DNA-microsatellites and the TTX-content of specimens from British Columbia (Prince Rupert area) and Alaska (Revillagigedo Island, Shelter Island, and Juneau) were analysed. TTX-concentrations were low in newts from BC and Revillagigedo Island, but high in specimens from mainland Juneau, which had been deliberately introduced from Shelter Island, where TTX was not detectable in the individuals sampled. No significant genetic differences were detected between these populations, which may correlate with the high intraspecies variability of TTX. It is still an open question, which factors favour or induce the toxin production in the newts.
Assuntos
Salamandridae , Animais , Humanos , América do Norte , Salamandridae/genética , TetrodotoxinaRESUMO
To date, there has been no published investigation on the trophic diversity in any tepui summit vertebrate. In this paper, we analyzed the dietary composition of a tepui summit endemic toad, Oreophrynella quelchii from Roraima-tepui, and compared it with that of O. nigra from Kukenán-tepui, to examine to what extent diet differs between these two sister species across isolated, although neighboring, tepui tops. The digestive tracts of a total of 197 toads were dissected: 111 from O. quelchii and 86 from O. nigra. The diet composition of O. quelchii was relatively diverse, with 13 major prey categories; mites (Acari, 36.5%) and beetles (Coleoptera, 21.0%) numerically dominated its diet. Despite occurring on two different tepui summits, O. quelchii and O. nigra exhibited a similar diet composition, although in O. nigra mites (Acari, 42.4%) and hymenopterans (especially ants, 16.9%) numerically dominated the diet. The present data suggest that tepui summit Oreophrynella species are flexible in their diet and are active foragers that also feed on aquatic arthropods, successful strategies in tepui competitive environments.
RESUMO
Tetrodotoxin (TTX) contents of wild-caught Chinese red-bellied newts, Cynops orientalis, and their offspring captive-reared from eggs to metamorphosed juveniles, were analysed using post-column LC-fluorescent detection (LC-FLD) and high resolution hydrophilic interaction liquid chromatography/mass spectrometry (HR-HILIC-LC/MS). TTX was detected in the parent newts and their eggs, but not in the larvae and juveniles raised under artificial condition over 20 months. However, juveniles reared in the presence of their parents, contained TTX-concentrations up to 8.05 µg/g. The origin of TTX may be implied from a close connection between the parents and their offspring.
Assuntos
Salamandridae , Animais , Cromatografia Líquida , Larva , Espectrometria de Massas , Tetrodotoxina/análise , Tetrodotoxina/toxicidadeRESUMO
Beetles of the genus Anthrenus are widespread worldwide and are quite common as grain and stock pests, especially in rural regions and suburban areas. The larvae of the beetles can trigger skin and occasionally mucosal reactions upon contact. The unfamiliar but typical image of "Anthrenus dermatitis" can easily be misinterpreted if beetle larvae are not detected or a causal link with them is not suspected. In the present article, the causes of Anthrenus dermatitis are discussed using as an example the larva of the woolly flower beetle (Anthrenus verbasci). For the examination of allergic causes, a prick-to-prick test with native larval material was performed in a patient and the specific IgE was determined by CAP coupling as well as in addition to a commercially available ImmunoCAP against larvae of a related beetle species. With the help of an immunoblot, an attempt was made to determine the responsible allergens in the native larval tissue. Important anatomical fine structures of the larva of a woolly herb flower beetle were analyzed using scanning electron microscopy to understand functional processes that lead to the clinical picture of Anthrenus dermatitis. Our allergological findings suggest an IgE-mediated, immediate-type allergy.
Assuntos
Besouros , Dermatite , Hipersensibilidade , Alérgenos , Animais , Humanos , Larva , Pele , Testes CutâneosRESUMO
Based on its mandibular gland secretion, the earless monitor lizard, Lanthanotus borneensis, has been considered a venomous animal like other members of the Toxicofera group, including Heloderma. In the present study, the gland structure and teeth of L. borneensis were examined by micro-tomography (µCT) and scanning electron microscopy (SEM), respectively, and proteomic analysis of the gland extract was performed. The mandibular gland consists of six compartments with separate ducts. The pleurodont teeth of the lower and upper jaw are not grooved but possess a sharp ridge on the anterior surface. Proteomic analysis of the gland extract confirmed previous studies that kallikrein enzymes are the major biologically active components. In view of the lizard's biology, its mandibular gland secretion is obviously not needed for prey capture or defence. It seems not justified the labelling of L. borneensis as a venomous animal. However, definitively answering this question requires toxinological studies on natural prey.
Assuntos
Lagartos , Peçonhas , Animais , Calicreínas , Proteômica , DenteRESUMO
Red-spotted newts, Notophthalmus viridescens, contain tetrodotoxin (TTX) and its analogue 6-epiTTX in variable concentrations. In a follow-up study, newts were sampled from a pond in Pennsylvania, USA, in 2010, 2014, and 2018. Their toxin levels were assayed by liquid-chromatography-fluorescence detection (LC-FLD), and assessment of their infection with endoparasites such as nematodes and helminths was performed by histological examination of internal organs. In the 2010 and 2014 samples, average prevalence of parasite infection was 53 and 60%, respectively, but reached 100% in the 2018 sample, where metacercaria stages of the digenean trematode genus Australapatemon/Apatemon (family: Strigeidae) were predominant causing severe tissue damage in liver and kidney. Mean values of TTX and 6-epiTTX were not significantly different in parasitized or parasite-free newts over the study period, confirming previous findings that host toxicity and parasite load are not negatively correlated. Whereas the role of TTX in defence against predators is undisputed, its efficacy to prevent parasitic infections is less obvious. Toxin-resistance of various metazoan parasites may promote their widespread occurrence in poisonous newts.
Assuntos
Antiplatelmínticos/uso terapêutico , Notophthalmus viridescens/parasitologia , Tetrodotoxina/uso terapêutico , Infecções por Trematódeos/veterinária , Animais , Notophthalmus viridescens/fisiologia , Pennsylvania , Substâncias Protetoras , Salamandridae , Trematódeos , Infecções por Trematódeos/tratamento farmacológicoRESUMO
Phlotoxin-1 (PhlTx1) is a peptide previously identified in tarantula venom (Phlogius species) that belongs to the inhibitory cysteine-knot (ICK) toxin family. Like many ICK-based spider toxins, the synthesis of PhlTx1 appears particularly challenging, mostly for obtaining appropriate folding and concomitant suitable disulfide bridge formation. Herein, we describe a procedure for the chemical synthesis and the directed sequential disulfide bridge formation of PhlTx1 that allows for a straightforward production of this challenging peptide. We also performed extensive functional testing of PhlTx1 on 31 ion channel types and identified the voltage-gated sodium (Nav) channel Nav1.7 as the main target of this toxin. Moreover, we compared PhlTx1 activity to 10 other spider toxin activities on an automated patch-clamp system with Chinese Hamster Ovary (CHO) cells expressing human Nav1.7. Performing these analyses in reproducible conditions allowed for classification according to the potency of the best natural Nav1.7 peptide blockers. Finally, subsequent in vivo testing revealed that intrathecal injection of PhlTx1 reduces the response of mice to formalin in both the acute pain and inflammation phase without signs of neurotoxicity. PhlTx1 is thus an interesting toxin to investigate Nav1.7 involvement in cellular excitability and pain.
Assuntos
Analgésicos/isolamento & purificação , Peptídeos/isolamento & purificação , Venenos de Aranha/química , Bloqueadores do Canal de Sódio Disparado por Voltagem/isolamento & purificação , Sequência de Aminoácidos , Analgésicos/química , Analgésicos/farmacologia , Analgésicos/uso terapêutico , Animais , Células CHO , Cricetulus , Feminino , Formaldeído , Camundongos Endogâmicos C57BL , Canal de Sódio Disparado por Voltagem NAV1.7/fisiologia , Oócitos , Dor/induzido quimicamente , Dor/tratamento farmacológico , Peptídeos/química , Peptídeos/farmacologia , Peptídeos/uso terapêutico , Dobramento de Proteína , Aranhas , Bloqueadores do Canal de Sódio Disparado por Voltagem/química , Bloqueadores do Canal de Sódio Disparado por Voltagem/farmacologia , Bloqueadores do Canal de Sódio Disparado por Voltagem/uso terapêutico , Xenopus laevisRESUMO
αδ-Bungarotoxins, a novel group of long-chain α-neurotoxins, manifest different affinity to two agonist/competitive antagonist binding sites of muscle-type nicotinic acetylcholine receptors (nAChRs), being more active at the interface of α-δ subunits. Three isoforms (αδ-BgTx-1-3) were identified in Malayan Krait (Bungarus candidus) from Thailand by genomic DNA analysis; two of them (αδ-BgTx-1 and 2) were isolated from its venom. The toxins comprise 73 amino acid residues and 5 disulfide bridges, being homologous to α-bungarotoxin (α-BgTx), a classical blocker of muscle-type and neuronal α7, α8, and α9α10 nAChRs. The toxicity of αδ-BgTx-1 (LD50 = 0.17-0.28â µg/g mouse, i.p. injection) is essentially as high as that of α-BgTx. In the chick biventer cervicis nerve-muscle preparation, αδ-BgTx-1 completely abolished acetylcholine response, but in contrast with the block by α-BgTx, acetylcholine response was fully reversible by washing. αδ-BgTxs, similar to α-BgTx, bind with high affinity to α7 and muscle-type nAChRs. However, the major difference of αδ-BgTxs from α-BgTx and other naturally occurring α-neurotoxins is that αδ-BgTxs discriminate the two binding sites in the Torpedo californica and mouse muscle nAChRs showing up to two orders of magnitude higher affinity for the α-δ site as compared with α-ε or α-γ binding site interfaces. Molecular modeling and analysis of the literature provided possible explanations for these differences in binding mode; one of the probable reasons being the lower content of positively charged residues in αδ-BgTxs. Thus, αδ-BgTxs are new tools for studies on nAChRs.
Assuntos
Bungarotoxinas/química , Bungarus , Proteínas de Peixes/química , Proteínas Musculares/química , Receptores Nicotínicos/química , Animais , Sítios de Ligação , Bungarotoxinas/metabolismo , Feminino , Proteínas de Peixes/metabolismo , Masculino , Camundongos , Proteínas Musculares/metabolismo , Receptores Nicotínicos/metabolismo , TorpedoRESUMO
Because of its bitter taste, quinine elicits strong antifeedant and toxic effects in animals including arthropods. In the present study, two mantis, Sphodromantis viridis, Hierodula membranacea, and two spider species, Nephila edulis, Selenocosmia javanensis, were offered a quinine solution or prey (crickets) contaminated or injected with quinine, which they ingested and survived without apparent toxic symptoms. Analysis of their faeces and, in the case of spiders, of silk from their web revealed that quinine was excreted over a period of 8-12 (mantids) or 7 days (spiders). Interestingly, the silk glands of the spiders served as an additional excretory organ of quinine. Both, mantids and spiders were shown to tolerate high amounts of quinine in their prey. Obviously, the bitter taste of this compound is not perceived by their gustatory receptors and consequently triggers no aversive reactions.
Assuntos
Mantódeos/fisiologia , Quinina , Aranhas/fisiologia , Animais , Fezes/química , Gryllidae , Mantódeos/metabolismo , Comportamento Predatório , Seda/química , Especificidade da Espécie , Aranhas/metabolismo , PaladarRESUMO
Frogs of the family Dendrobatidae are known to contain toxic alkaloids in their skin secretion, but Allobates species of the closely related Aromobatidae family are considered to lack toxic secretions. However, contradictory results have been reported. Analyses of alcohol extracts from three different Allobates species from South-America (Guiana Shield), Central America (Costa Rica), and from the dendrobatid frog Silverstoneia flotator confirm the absence of alkaloids and tetrodotoxin in aromobatids and in a dendrobatid of the subfamily Colostethinae.
Assuntos
Alcaloides/análise , Anuros , Tetrodotoxina/análise , Animais , Costa Rica , Guiana , Pele/químicaRESUMO
Toads of the genus Melanophryniscus contain toxic alkaloids sequestered from a diet of mainly ants and mites. Wild-caught specimens of M. klappenbachi were fed an alkaloid-free diet and their alkaloid profile was analyzed during 38 months in captivity. Individual alkaloid patterns varied considerably. Over time, the concentration of two alkaloids, pumiliotoxin 251D and 3,5-disubstituted indolizidine 195B, significantly declined, suggesting that the toads depend on continuous access to alkaloid-containing prey to maintain natural levels of their chemical defense.
Assuntos
Alcaloides/administração & dosagem , Alcaloides/metabolismo , Ração Animal/análise , Bufonidae/fisiologia , Dieta/veterinária , Pele/metabolismo , Alcaloides/química , Animais , Estrutura MolecularRESUMO
For the first time, alcohol extracts of Atelopus hoogmoedi from the Guiana Shield in Suriname and Guyana were analyzed for the presence of tetrodotoxin (TTX) and of its analogues by high resolution hydrophilic interaction liquid chromatography/mass spectrometry. One specimen from Suriname was found to contain TTX and 4-epiTTX. Using a monoclonal antibody-based immunohistochemical staining technique, TTX was localized mainly in the granular glands and epithelium of the skin, but not in internal organs except liver showing weak TTX-positive reaction. In two specimens collected in Guyana, none of the toxins were detected.
Assuntos
Bufonidae/fisiologia , Tetrodotoxina/metabolismo , Distribuição Animal , Animais , América do SulRESUMO
To cope with toxic metabolites plants use for defence, herbivorous insects employ various adaptive strategies. For oviposition, the fruit fly Dacus siliqualactis (Tephritidae) uses milkweed plants of the genus Gomphocarpus (Asclepiadaceae) by circumventing the plant's physical (gluey latex) and chemical (toxic cadenolides) defence. With its long, telescope-like ovipositor, the fly penetrates the exo- and endocarp of the fruit and places the eggs on the unripe seeds located in the centre of the fruit. Whereas most plant parts contain high concentrations of cardenolides such as gomphoside, calotropin/calacatin and gomphogenin, only the seeds exhibit low cardenolide levels. By surmounting physical barriers (fruit membranes, latex), the fly secures a safe environment and a latex-free food source of low toxicity for the developing larvae. One amino acid substitution (Q111V) at the cardenolide binding site of the fly's Na+, K+-ATPase was detected, but the significance of that substitution: reducing cardenolide sensitivity or not, is unclear. However, poisoning of the larvae by low levels of cardenolides is assumed to be prevented by non-resorption and excretion of the polar cardenolides, which cannot passively permeate the midgut membrane. This example of an insect-plant interaction demonstrates that by morphological and behavioural adaptation, a fruit fly manages to overcome even highly effective defence mechanisms of its host plant.
Assuntos
Apocynaceae/parasitologia , Oviposição , Tephritidae/fisiologia , Animais , Apocynaceae/anatomia & histologia , Apocynaceae/química , Cardenolídeos/análise , Feminino , Frutas/anatomia & histologia , Frutas/parasitologia , Interações Hospedeiro-Parasita , Larva/fisiologia , Látex , Análise de Sequência de DNA , Análise de Sequência de Proteína , ATPase Trocadora de Sódio-Potássio/química , Tephritidae/crescimento & desenvolvimentoRESUMO
Salamanders have evolved a wide variety of antipredator mechanisms and behavior patterns, including toxins and noxious or adhesive skin secretions. The high bonding strength of the natural bioadhesives makes these substances interesting for biomimetic research and applications in industrial and medical sectors. Secretions of toxic species may help to understand the direct effect of harmful substances on the cellular level. In the present study, the biocompatibility of adhesive secretions from four salamander species (Plethodon shermani, Plethodon glutinosus, Ambystoma maculatum, Ambystoma opacum) were analyzed using the MTT assay in cell culture and evaluated against toxic secretions of Pleurodeles waltl, Triturus carnifex, Pseudotriton ruber, Tylototriton verrucosus, and Salamandra salamandra. Their effect on cells was tested in direct contact (direct culture) or under the influence of the extract (indirect exposure) in accordance with the protocol of the international standard norm ISO 10993-5. Human dermal fibroblasts (NHDF), umbilical vein endothelial cells (HUVEC), and articular chondrocytes (HAC), as well as the cell lines C2C12 and L929 were used in both culture types. While the adhesive secretions from Plethodon shermani are cytocompatible and those of Ambystoma opacum are even advantageous, those of Plethodon glutinosus and Ambystoma maculatum appear to be cytotoxic to NDHF and HUVEC. Toxic secretions from Salamandra salamandra exhibited harmful effects on all cell types. Pseudotriton ruber and Triturus carnifex secretions affected certain cell types marginally; those from Pleurodeles waltl and Tylototriton verrucosus were generally well tolerated. The study shows for the first time the effect of salamander secretions on the viability of different cell types in culture. Two adhesive secretions appeared to be cell compatible and are therefore promising candidates for future investigations in the field of medical bioadhesives. Among the toxic secretions tested, only two of the five had a harmful effect on cells, indicating different cell toxicity mechanisms.