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J. Venom. Anim. Toxins incl. Trop. Dis.

V.20, 2014.

Original paper - ISSN 1678-9199.

 

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


Sandro Rostelato-Ferreira1*, Cháriston A Dal Belo2, Gildo B Leite1, Stephen Hyslop1, Léa Rodrigues-Simioni1

 

1Departamento de Farmacologia, Faculdade de Ciências Médicas, Universidade Estadual de Campinas (UNICAMP), CP 6111, Campinas, SP 13083-970, Brasil

2Laboratório de Neurobiologia e Toxinologia, (LANETOX), Universidade Federal do Pampa, (UNIPAMPA), Av. Antônio Trilha, 1847, Centro, CEP 97300-000 São Gabriel, RS, Brazil

 

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.

 

Key words: Neurotransmitter release; Ouabain; Presynaptic; Rhinella schneideri; Toad venom

 

Ethics committee approval

The present study was approved by the institutional Committee for Ethics in Animal Use (CEUA/UNICAMP, protocol no. 1552–1) and was performed in accordance with the ethical guidelines established by the Brazilian Society of Laboratory Animal Science (SBCAL, formerly the Brazilian College of Animal Experimentation – COBEA).

 

Received: January 17, 2014; Accepted: June 30, 2014

 

Corresponding author: sandrorostelato@yahoo.com.br

 

Competing interests

The authors declare that there are no competing interests.

 

Authors’ contributions

SRF and LRS are the lead researchers of this study. SRF, CADB and LRS designed the study. SRF and GBL performed experiments and their analysis. SRF, CADB, SH and LRS participated in the analysis of the results and article writing. All authors read and approved the final manuscript.