Presynaptic Activity of an Isolated Fraction from Rhinella schneideri Poison

Purpose: Rhinella schneideri is a toad found in many regions of the South America. The poison of the glands has cardiotoxic effect in animals and neuromuscular effects in mice and avian preparation. The purpose of this work was to identify the toxin responsible for the neuromuscular effect in avian...

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Main Authors: Sandro Rostelato-Ferreira, Cháriston André Dal Belo, Pedro Ismael da Silva Junior, Stephen Hyslop, Léa Rodrigues-Simioni, Thomaz Augusto Alves Rocha-e-Silva
Format: Article
Language:English
Published: Tabriz University of Medical Sciences 2018-08-01
Series:Advanced Pharmaceutical Bulletin
Subjects:
Online Access:http://apb.tbzmed.ac.ir/PDF/apb-8-517.pdf
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author Sandro Rostelato-Ferreira
Cháriston André Dal Belo
Pedro Ismael da Silva Junior
Stephen Hyslop
Léa Rodrigues-Simioni
Thomaz Augusto Alves Rocha-e-Silva
author_facet Sandro Rostelato-Ferreira
Cháriston André Dal Belo
Pedro Ismael da Silva Junior
Stephen Hyslop
Léa Rodrigues-Simioni
Thomaz Augusto Alves Rocha-e-Silva
author_sort Sandro Rostelato-Ferreira
collection DOAJ
description Purpose: Rhinella schneideri is a toad found in many regions of the South America. The poison of the glands has cardiotoxic effect in animals and neuromuscular effects in mice and avian preparation. The purpose of this work was to identify the toxin responsible for the neuromuscular effect in avian and mice neuromuscular preparation. Methods: The methanolic extract from R. schneideri poison was fractioned by reversed phase HPLC. The purity and molecular mass were determined by LC/MS mass spectrometry. Chick biventer cervicis and mouse phrenic-nerve diaphragm were used as neuromuscular preparations to identify the toxin. Results: The purification resulted in 32 fractions, which 4 of them were active in neuromuscular preparation. The toxin of fraction 20 were chosen for better reproducibility of the whole extract activity and its molecular mass was 730.6 Da. The toxin produced facilitation of the muscle contraction followed by a complete neuromuscular blockade in chick biventer cervicis preparation in 90 min without interfering with the exogenous response to ACh and KCl. The quantal content was increased from 128 ± 13 (control) to 216 ± 44 (after 5 min and sustained until 60 min) in the presence of the toxin. Conclusion: In conclusion, our results demonstrated that the neuromuscular action of the poison of Rhinella schneideri is a multitoxin effect. More, the present work first isolated a 730.6 Da toxin that better represent the whole poison neuromuscular effect, to which is attributed a presynaptic action in avian and mouse neuromuscular preparation.
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spelling doaj.art-2790089f888d4934958afb3f199000bd2022-12-22T01:08:03ZengTabriz University of Medical SciencesAdvanced Pharmaceutical Bulletin2228-58812251-73082018-08-018351752210.15171/apb.2018.060apb-19511Presynaptic Activity of an Isolated Fraction from Rhinella schneideri PoisonSandro Rostelato-Ferreira0Cháriston André Dal Belo1Pedro Ismael da Silva Junior2Stephen Hyslop3Léa Rodrigues-Simioni4Thomaz Augusto Alves Rocha-e-Silva5Departamento de Farmacologia, Faculdade de Ciências Médicas, Universidade Estadual de Campinas (UNICAMP), Zip Code 13083-970, Campinas, SP, Brazil.Centro de Ciências Rurais de São Gabriel, Universidade Federal do Pampa (UNIPAMPA), São Gabriel, RS, Brazil.Laboratório Especial de Toxinologia Aplicada, Instituto Butantan, São Paulo, SP, Brazil.Departamento de Farmacologia, Faculdade de Ciências Médicas, Universidade Estadual de Campinas (UNICAMP), Zip Code 13083-970, Campinas, SP, Brazil.Departamento de Farmacologia, Faculdade de Ciências Médicas, Universidade Estadual de Campinas (UNICAMP), Zip Code 13083-970, Campinas, SP, Brazil.School of Medicine Faculdade Israelita de Ciências da Saúde Albert Einstein, São Paulo, SP, Brazil.Purpose: Rhinella schneideri is a toad found in many regions of the South America. The poison of the glands has cardiotoxic effect in animals and neuromuscular effects in mice and avian preparation. The purpose of this work was to identify the toxin responsible for the neuromuscular effect in avian and mice neuromuscular preparation. Methods: The methanolic extract from R. schneideri poison was fractioned by reversed phase HPLC. The purity and molecular mass were determined by LC/MS mass spectrometry. Chick biventer cervicis and mouse phrenic-nerve diaphragm were used as neuromuscular preparations to identify the toxin. Results: The purification resulted in 32 fractions, which 4 of them were active in neuromuscular preparation. The toxin of fraction 20 were chosen for better reproducibility of the whole extract activity and its molecular mass was 730.6 Da. The toxin produced facilitation of the muscle contraction followed by a complete neuromuscular blockade in chick biventer cervicis preparation in 90 min without interfering with the exogenous response to ACh and KCl. The quantal content was increased from 128 ± 13 (control) to 216 ± 44 (after 5 min and sustained until 60 min) in the presence of the toxin. Conclusion: In conclusion, our results demonstrated that the neuromuscular action of the poison of Rhinella schneideri is a multitoxin effect. More, the present work first isolated a 730.6 Da toxin that better represent the whole poison neuromuscular effect, to which is attributed a presynaptic action in avian and mouse neuromuscular preparation.http://apb.tbzmed.ac.ir/PDF/apb-8-517.pdfNeurotransmitter releasePresynaptic toxinNeuromuscular junctionIsolated fraction
spellingShingle Sandro Rostelato-Ferreira
Cháriston André Dal Belo
Pedro Ismael da Silva Junior
Stephen Hyslop
Léa Rodrigues-Simioni
Thomaz Augusto Alves Rocha-e-Silva
Presynaptic Activity of an Isolated Fraction from Rhinella schneideri Poison
Advanced Pharmaceutical Bulletin
Neurotransmitter release
Presynaptic toxin
Neuromuscular junction
Isolated fraction
title Presynaptic Activity of an Isolated Fraction from Rhinella schneideri Poison
title_full Presynaptic Activity of an Isolated Fraction from Rhinella schneideri Poison
title_fullStr Presynaptic Activity of an Isolated Fraction from Rhinella schneideri Poison
title_full_unstemmed Presynaptic Activity of an Isolated Fraction from Rhinella schneideri Poison
title_short Presynaptic Activity of an Isolated Fraction from Rhinella schneideri Poison
title_sort presynaptic activity of an isolated fraction from rhinella schneideri poison
topic Neurotransmitter release
Presynaptic toxin
Neuromuscular junction
Isolated fraction
url http://apb.tbzmed.ac.ir/PDF/apb-8-517.pdf
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