Dataset Information

Characterization of extracellular vesicles from Bothrops jararaca snake venom

ABSTRACT: Venoms are important evolutionary keys and innovation for several animal taxons., including snakes. Their use is specially related to feed, predation and defense. Snake venoms are composed mainly by free secreted proteins, about 98%, and stored in the lumen of the venom gland, where they are processed through poorly understood mechanisms. In the perspective of protein-diversity, venoms undergo evolutionary pressure which generate a rapid evolutionary response, causing a great diversity in their toxin-components. We know now that extracellular vesicles exist in snake venom, although their biological role are still unknown. We believe that understanding EV-mediated effects could change our way of seeing envenoming, especially long-term sequelae. From what we know about EVs and snake venoms, there is a lot of potential of cross-organism communication occurrence between snakes and human victims. To advance in the comprehension of venom EVs function, we used fresh B. jararaca venom as our model. Fresh venom was fractionated by sequential centrifugation, resulting in two populations of vesicles (Bj-EVs). Purified Bj-EVs were analyzed by electron microscopy, NTA and proteomics. The interaction of Bj-EVs with mammalian cells was accessed by fluorescence and electron microscopy.

INSTRUMENT(S): LTQ Orbitrap Velos

ORGANISM(S): Bothrops Jararaca (jararaca) (bothrops Jajaraca)

TISSUE(S): Venom

SUBMITTER: Larissa Gonçalves Machado

LAB HEAD: Russolina Benedeta Zingali

PROVIDER: PXD037375 | Pride | 2023-03-10

REPOSITORIES: Pride

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Publications

Extracellular Vesicles from <i>Bothrops jararaca</i> Venom Are Diverse in Structure and Protein Composition and Interact with Mammalian Cells.

Gonçalves-Machado Larissa L Verçoza Brunno Renato Farias BRF Nogueira Fábio César Sousa FCS Melani Rafael Donadélli RD Domont Gilberto Barbosa GB Rodrigues Silas Pessini SP Rodrigues Juliany Cola Fernandes JCF Zingali Russolina Benedeta RB

Toxins 20221119 11

Snake venoms are complex cocktails of non-toxic and toxic molecules that work synergistically for the envenoming outcome. Alongside the immediate consequences, chronic manifestations and long-term sequelae can occur. Recently, extracellular vesicles (EVs) were found in snake venom. EVs mediate cellular communication through long distances, delivering proteins and nucleic acids that modulate the recipient cell's function. However, the biological roles of snake venom EVs, including possible cross- ...[more]

PMID: 36422980

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Project description:Snake venoms are complex protein mixtures with different biological activities that can act in both their preys and human victims. Many of these proteins play a role in prey capture and in the digestive process of these animals. It is known that some snakes are resistant to the toxicity of their own venom by mechanisms not yet fully elucidated. However, it was observed in the Laboratory of Herpetology of Instituto Butantan that some Bothrops moojeni individuals injured by the same snake species showed mortalities caused by envenoming effects. This study analyzed the biochemical composition of 13 venom and plasma samples from Bothrops moojeni specimens to assess differences in their protein composition. Application of sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) showed distinct venom protein profiles, but very homogeneous plasma profiles. Western Blotting (WB) was performed with plasma samples, which were submitted to incubation with the respective venom. Some individuals showed an immunorecognized band zone around 25 kDa, indicating interaction between the same individual plasma and venom proteins. Crossed-WB assay using non-self-plasma and venom showed that this variability is due to venom protein composition instead of plasma composition. These venoms presented higher caseinolytic, collagenolytic and coagulant activities than the venoms without these regions recognized by WB. Mass spectrometry analysis revealed that these individuals present, in addition to higher protein concentrations, other exclusive proteins in their composition. When these samples were tested in vivo, the results also showed higher lethality in these venoms, but lower hemorrhagic activity than in the venoms without these regions recognized by WB. In conclusion, some Bothrops moojeni specimens differ in venom composition, which may have implications in envenomation. Moreover, the high individual venom variability found in this species demonstrates the importance to work with individual analysis in studies involving intraspecific venom variability and venom evolution.

Project description:Bothrops moojeni has a wide distribution in Brazil and represents a serious medical concern at some localities. Previous works reported that the manifestations of snakebites caused by B. moojeni juveniles were mainly related to coagulation, while those caused by adults had more prominent local damage. In this work, we aimed to analyze the venoms of B. moojeni at different life stages to better understand how the venom shifts along the ontogeny. Snakes were grouped by age and sex, and venom pools were formed accordingly. Compositional analyses by one-dimensional electrophoresis (1-DE), chromatography and mass spectrometry revealed that ontogenetic changes may be mostly related to phospholipase A2 (PLA2) and metalloproteases. Regarding the functional aspect of the venoms, in vitro assays of proteolytic, L-amino acid oxidase, PLA2 and coagulant activities were assayed, but only the first and the last showed age-related changes, with the venom of younger snakes displaying lower proteolytic and higher coagulant activities up to 1 year-old; from 2 years-old onward, the venoms presented the opposite relation. The venoms of 3 years-old snakes were exceptions to the compositional and functional pattern of adults. Sex-related differences were observed at specific groups and not related to age. In vivo experiments (median lethal dose and hemorrhagic activity) were statistically similar between neonates and adults, but observation of the time each venom took to kill the mice revealed that the adult one seemed to act faster than the venoms of the neonates. All venoms were mostly recognized by the antibothropic serum and displayed similar profiles to 1-DE in western blotting. In conclusion, the venoms showed ontogenetic shift in composition and activities. Furthermore, this change occurred from 1 to 2 years-old, and interestingly the 3 years-old pools had particular characteristics, which highlight the importance of extending the studies to better understand venom variability.

Dataset Information

Characterization of extracellular vesicles from Bothrops jararaca snake venom

Publications

Extracellular Vesicles from <i>Bothrops jararaca</i> Venom Are Diverse in Structure and Protein Composition and Interact with Mammalian Cells.

Similar Datasets

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