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      Presynaptic neuromuscular action of a methanolic extract from the venom of Rhinella schneideri toad

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          Abstract

          Background

          Rhinella schneideri, previously known as Bufo paracnemis, is a common toad in many regions of Brazil. Its venom exerts important cardiovascular effects on humans and other animals. Although this toad venom has been the subject of intense investigations, little is known about its neuromuscular activity.

          Methods

          The neurotoxicity of a methanolic extract of R. schneideri venom was tested on mouse phrenic nerve-diaphragm (PND) preparations mounted for conventional twitch tension recording – in response to indirect stimulation – and for electrophysiological measurements.

          Results

          Venom extract (50 μg/mL) increased the muscle twitch tension in PND preparations but did not significantly alter the resting membrane potential values. Electrophysiological evaluations showed that the extract (50 μg/mL) significantly augmented the frequency of miniature end-plate potential (from 38 ± 3.5 to 88 ± 15 after 60 minutes; n = 5; p < 0.05) and quantal content (from 128 ± 13 to 272 ± 34 after five minutes; n = 5; p < 0.05). Pretreatment with ouabain (1 μg/mL) for five minutes prevented the increase in quantal content (117 ± 18 and 154 ± 33 after five and 60 minutes, respectively).

          Conclusion

          These results indicate that the methanolic extract of R. schneideri venom acts primarily presynaptically to enhance neurotransmitter release in mouse phrenic-diaphragm preparations.

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          Most cited references40

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          An analysis of the end-plate potential recorded with an intracellular electrode.

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            Pharmacology and toxicology of toad venom.

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              Comparison of toad venoms from different Bufo species by HPLC and LC-DAD-MS/MS.

              Toad venom, called Chansu in China, has been widely used for the treatment of heart failure, sores, pains, and various cancers for a long time in clinic. The aim of the study is to investigate the chemical differences among a variety of toad venoms from different geographic locations and related Bufo species. Ten batches of commercial toad venom collected from different regions in China, one batch of fresh toad venom obtained from Bufo bufo gargarizans, and six batches of related Bufo species were analyzed by HPLC and LC-DAD-MS/MS. Individual components were identified by comparison of retention times, UV spectra, and mass spectra with authentic compounds, standard addition, as well as summarized MS fragmentation rules. Based on the profile of identified constituents and the content of cinobufagin and resibufogenin, the chemical differences observed among different samples are discussed. Overall, 43 compounds were identified in the methanolic extracts of the different samples of toad venom. Besides of suberoyl arginine, several free bufadienolides, bufadienolide sulfates, and suberoyl esters of bufadienolides were found. The total amounts of cinobufagin and resibufogenin, which are the only two control markers according to the current Chinese Pharmacopoeia, varied widely from 0.7% to 10.9% in the commercial Chansu samples collected in the different locations in China. Low levels of resibufogenin, but no cinobufagin was observed in the samples from Bufo melanosticus and Bufo marinus, and even neither of both compounds was found in the sample from Bufo viridis. The chemical profiles of the different commercial and collected toad venoms from related Bufo species differed significantly, not only in the absolute and relative contents, but also in the number and type of the constituents. The main reason for this variation are species-specific differences, but additional factors, such as the harvest and post-harvest processing, and adaption to environmental factors in different geographic locations, also seem to contribute. Copyright 2010 Elsevier Ireland Ltd. All rights reserved.
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                Author and article information

                Contributors
                Journal
                J Venom Anim Toxins Incl Trop Dis
                J Venom Anim Toxins Incl Trop Dis
                The Journal of Venomous Animals and Toxins Including Tropical Diseases
                BioMed Central
                1678-9199
                2014
                4 July 2014
                : 20
                : 30
                Affiliations
                [1 ]Departamento de Farmacologia, Faculdade de Ciências Médicas, Universidade Estadual de Campinas (UNICAMP), CP 6111, Campinas, SP 13083-970, Brasil
                [2 ]Laboratório de Neurobiologia e Toxinologia, (LANETOX), Universidade Federal do Pampa, (UNIPAMPA), Av. Antônio Trilha, 1847, Centro, CEP 97300-000 São Gabriel, RS, Brazil
                Article
                1678-9199-20-30
                10.1186/1678-9199-20-30
                4094905
                25024696
                394ffd0a-0a1d-43cc-8a55-993bd9ab4098
                Copyright © 2014 Rostelato-Ferreira et al.; licensee BioMed Central Ltd.

                This is an Open Access article distributed under the terms of the Creative Commons Attribution License ( http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver ( http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

                History
                : 17 January 2014
                : 30 June 2014
                Categories
                Research

                neurotransmitter release,ouabain,presynaptic,rhinella schneideri,toad venom

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