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Physiological and biochemical analysis of L. tredecimguttatus venom collected by electrical stimulation

Propiedades bioquímicas y fisiológicas del veneno del escorpión L. tredecimguttatus obenidor por estimulación eléctrica

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Abstract

The L. tredecimguttatus venom was collected by electrical stimulation and systematically analyzed. Gel electrophoresis and RP-HPLC showed that the venom consisted primarily of proteins with molecular weights above 10 kDa, most of which were high-molecular-mass acidic proteins, with fewer proteins and peptides below 10 kDa. The most abundant proteins in the venom were concentrated at around 100 kDa, which included latrotoxins- the principal toxic components of the venom. Injection of the venom in mice and cockroaches P. americana gave rise to obvious poisoned symptoms, with LD50 values of 0.16 mg/kg and 1.87 μg/g, respectively. Electrophysiological experiments showed that the venom could block the neuromuscular transmission in isolated mouse phrenic nerve-hemidiaphragm and rat vas deferens preparations. The low-molecular-weight fraction (<10 kDa) of the venom had no effect on the transmission. Enzymatic analysis indicated that the venom possess activities of several kinds of hydrolases including hyaluronidase and proteases. These results demonstrated that L. tredecimguttatus venom was basically a large-protein-constituted venom and is one of the most poisonous spider venoms known in the world. The mammalian toxicity of the venom was based on its larger proteins rather than on smaller proteins and peptides, and its hydrolase activities might be involved in the latrodectism. The use of electrical stimulation method to collect the venom has the advantages of avoiding contamination and repeated use of the valuable L. tredecimguttatus venom resources.

Resumen

En el presenta trabajo, se analiza el veneno de Latrodectus tredecimguttatus obtenido por estimulación eléctrica, método de recolección que evita la contaminación. Tras aplicación de técnicas de HPLC, espectrometría de masas y electroforesis en gel se deduce que el veneno contiene predominantemente proteínas ácidas de peso molecular superior a 10kDa. Las mas abundantes se concentran alrededor de 100 kDa, entre las que se encuentra la latrocina, principal componente tóxico del veneno. El análisis enzimático del veneno indica que incluye actividad enzimática de varios tipos de hidrolasas, incluyendo hialuronidasa y proteasas. Tras inyección del veneno en el ratón y en la cucaracha Periplaneta americana se obtuvieron valores de LD50 de 0,16 mg/kg y 1,87 microg/g respectivamente. Los experimentos electrofisiológicos mostraron que el veneno bloqueaba la transmisión neuromuscular tanto en preparado nervio frénico-hemidiafragma de ratón como en el de vas deferens de rata. La fracción del veneno que contenía péptidos de bajo peso molecular (inferior a 10 kDa) era inefectiva sobre la transmisión. En suma, los resultados muestran que la gran toxicidad del veneno de Latrodectus tredecimguttatus se relaciona con su contenido en proteínas de elevado peso molecular, cuyas actividades hidrolíticas están incluidas en el lactrodectismo.

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Authors and Affiliations

  1. College of Life Sciences, Hunan Normal University, 410081, Changsha, Hunan, China

    X. C. Wang, Z. G. Duan, J. Yang, X. J. Yan, H. Zhou, X. Z. He & S. P. Liang

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  1. X. C. Wang
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  2. Z. G. Duan
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Correspondence to S. P. Liang.

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Wang, X.C., Duan, Z.G., Yang, J. et al. Physiological and biochemical analysis of L. tredecimguttatus venom collected by electrical stimulation. J. Physiol. Biochem. 63, 221–230 (2007). https://doi.org/10.1007/BF03165785

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  • DOI: https://doi.org/10.1007/BF03165785

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