Archives of Toxicology

, Volume 93, Issue 7, pp 2065–2086 | Cite as

Neurotoxicity of Micrurus lemniscatus lemniscatus (South American coralsnake) venom in vertebrate neuromuscular preparations in vitro and neutralization by antivenom

  • Rafael S. Floriano
  • Raphael Schezaro-Ramos
  • Nelson J. SilvaJr.
  • Fábio Bucaretchi
  • Edward G. Rowan
  • Stephen HyslopEmail author


We investigated the effect of South American coralsnake (Micrurus lemniscatus lemniscatus) venom on neurotransmission in vertebrate nerve–muscle preparations in vitro. The venom (0.1–30 µg/ml) showed calcium-dependent PLA2 activity and caused irreversible neuromuscular blockade in chick biventer cervicis (BC) and mouse phrenic nerve–diaphragm (PND) preparations. In BC preparations, contractures to exogenous acetylcholine and carbachol (CCh), but not KCl, were abolished by venom concentrations ≥ 0.3 µg/ml; in PND preparations, the amplitude of the tetanic response was progressively attenuated, but with little tetanic fade. In low Ca2+ physiological solution, venom (10 µg/ml) caused neuromuscular blockade in PND preparations within ~ 10 min that was reversible by washing; the addition of Ca2+ immediately after the blockade temporarily restored the twitch responses, but did not prevent the progression to irreversible blockade. Venom (10 µg/ml) did not depolarize diaphragm muscle, prevent depolarization by CCh, or cause muscle contracture or histological damage. Venom (3 µg/ml) had a biphasic effect on the frequency of miniature end-plate potentials, but did not affect their amplitude; there was a progressive decrease in the amplitude of evoked end-plate potentials. The amplitude of compound action potentials in mouse sciatic nerve was unaffected by venom (10 µg/ml). Pre-incubation of venom with coralsnake antivenom (Instituto Butantan) at the recommended antivenom:venom ratio did not neutralize the neuromuscular blockade in PND preparations, but total neutralization was achieved with a tenfold greater volume of antivenom. The addition of antivenom after 50% and 80% blockade restored the twitch responses. These results show that M. lemniscatus lemniscatus venom causes potent, irreversible neuromuscular blockade, without myonecrosis. This blockade is apparently mediated by pre- and postsynaptic neurotoxins and can be reversed by coralsnake antivenom.


Antivenom Coralsnake venom Neuromuscular blockade α-Neurotoxin Neutralization Phospholipase A2 (β-neurotoxin) 



RSF was supported by a post-doctoral fellowship from Fundação de Amparo à Pesquisa do Estado de São Paulo—Brasil (FAPESP, Grant No. 2014/24409-8) and RSR was supported by a PhD studentship from Coordenação de Aperfeiçoamento de Pessoal de Nível Superior—Brasil (CAPES, Grant No. 02-P-4572/2018, Finance code 001). NJS and SH are supported by research fellowships from Conselho Nacional de Desenvolvimento Científico e Tecnológico—Brasil (CNPq, Grant Nos. 309320/2016-0 and 310547/2014-8, respectively).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest with this work.

Ethical approval

The relevant international, national, and/or institutional guidelines for the care and use of animals were followed in this work, as indicated in the section ‘animals’ of the methods.

Supplementary material

204_2019_2476_MOESM1_ESM.docx (10.6 mb)
Supplementary material 1 (DOCX 10880 kb)


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Authors and Affiliations

  1. 1.Departamento de Farmacologia, Faculdade de Ciências MédicasUniversidade Estadual de Campinas (UNICAMP)CampinasBrazil
  2. 2.Programa de Pós-Graduação em Ciências Ambientais e Saúde, Escola de Ciências Médicas, Farmacêuticas e BiomédicasPontifícia Universidade Católica de Goiás (PUC-GO)GoiâniaBrazil
  3. 3.Departamento de Pediatria e Centro de Informação e Assistência Toxicológica de Campinas (CIATox), Faculdade de Ciências MédicasUniversidade Estadual de Campinas (UNICAMP)CampinasBrazil
  4. 4.Strathclyde Institute of Pharmacy and Biomedical SciencesUniversity of StrathclydeGlasgowUK

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