A new crotamine-like from the rattlesnake (Crotalus durissus cumanensis) venom causing damages: Qualitative and Quantitative Cytotoxic Studies on subcellular and neuromuscular structures

  • Estefanie García Universidad Central de Venezuela, Faculty of Sciences, Electron Microscopy Centre. Caracas, Bolivarian Republic of Venezuela
  • Hector José Finol† Universidad Central de Venezuela, Faculty of Sciences, Electron Microscopy Centre. Caracas, Bolivarian Republic of Venezuela
  • Roschman González Universidad Central de Venezuela, Faculty of Sciences, Electron Microscopy Centre. Caracas, Bolivarian Republic of Venezuela
  • Alexis Rodríguez-Acosta Universidad Central de Venezuela, Anatomical Institute, Laboratory of Immunochemistry and Ultrastructure. Caracas, Bolivarian Republic of Venezuela https://orcid.org/0000-0003-1234-7522
Keywords: Crotamine-like peptide, Crotalus durissus cumanensis, mitochondria, motor- endplate, myotoxin, rattlesnake, smooth endoplasmic reticulum

Abstract

A quantitative model to expose the adrenal gland sub-cellular alterations produced by crotamine-like (C-L) from rattlesnake venom during 3, 6 and 24 hours (h), and also qualitative changes on mice neuromuscular structures in vivo were observed and calculated by transmission electron microscopy. A pure crotamine-like (C-L) isoform was obtained using a cationic exchange chromatography column from the rattlesnake Crotalus durissus cumanensis venom. The C-L SDS-PAGE (15.5 %) under non-reduced conditions exhibited a molecular mass of ~3 kDa single band. The C-L in vivo qualitative experiments induced ultrastructural changes in mouse neuromuscular structures at 3, 6 and 24 h, such as reduction in the number of acetylcholine vesicles, disorganisation of the secondary synaptic clefts, enlargement of the sub-sarcolemma space and alteration of the mitochondria morphology, number and cristae. Regarding neurotoxic actions in vivo, the animals injected with C-L presented spastic paralysis of the hind limbs. The quantitative alterations studied on the capillaries, the nucleus, the mitochondria the lipid inclusions, and the smooth endoplasmic reticulum were observed from 3 to 24 h after C-L injection. As far as it is known from the literature review, there are no quantitative records of similar sub-cellular alterations caused by crotamine.

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References

AGUILAR, I.; GUERRERO, B.; SALAZAR, A.M.; GIRÓN, M.E.; PÉREZ, J.C.; SÁNCHEZ, E.E.; RODRÍGUEZ-ACOSTA, A. Individual venom variability in the South American rattlesnake Crotalus durissus cumanensis. Toxicon. 50: 214-224. 2007.

REIS, L.P.G.; BOTELHO, A.F.M.; NOVAIS, C.R.; FIÚZA, A.T.L.; BARRETO, M.S.O.; FERREIRA, M.G.; BONILLA, C.; CHAVEZ-OLÓRTEGUI, C.; MELO, M.M. Cardiotoxic Effects of Micrurus surinamensis (Cuvier, 1817) Snake Venom. Cardiovasc. Toxicol. 21(6): 462-471. 2021.

MACIEL, F.V.; RAMOS-PINTO, Ê.K.; VALÉRIO-SOUZA, N.M.; GONÇALVES DE A., T.A.; ORTOLANI, P.L.; FORTES-DIAS, C.L.; GARRIDO – CAVALCANTE, W.L. Varespladib (LY315920) prevents neuromuscular blockage and myotoxicity induced by crotoxin on mouse neuromuscular preparations. Toxicon. 30: 40-45. 2021.

CHANG, C.; TSENG, K. Effect of crotamine, a toxin of South American rattlesnake venom on the sodium channel of murine skeletal muscle. Br. J. Pharmacol. 63: 551-559. 1978.

OGUIURA, N.; BONI-MITAKE, M.; RÁDIS-BAPTISTA, G. New view on crotamine, a small basic polypeptide myotoxin from South American rattlesnake venom. Toxicon. 46(4): 363-370. 2005.

FISHER, R.A. Statistical Methods for Research Workers. Breakthroughs in Statistics. In: Kotz, S.; Johnson, N.L. (Eds.). Springer Series in Statistics. Springer New York, NY. 601 pp. 1992.

GINEBAUGH, S.P.; CYPHERS, E.D.; LANKA, V.; ORTIZ, G.; MILLER, E.W.; LAGHAEI, R.; MERINEY, S.D. The frog motor nerve terminal has very brief action potentials and three electrical regions predicted to differentially control transmitter release. J. Neurosci. 40: 3504-3516. 2020.

GIRÓN, M.E.; PINTO, A.; FINOL, H.; AGUILAR, I.; RODRÍGUEZ-ACOSTA, A. Kidney structural and ultrastructural pathological changes induced by uracoan rattlesnake (Crotalus vegrandis Klauber 1941) venom. J. Submicrosc. Cytol. Pathol. 34: 447-459. 2002.

GONÇALVES, J.M.; ARANTES, E.G. Estudos sobre venenos de serpentes brasileiras III—Determinação quantitativa de crotamina no veneno de cascavel Brasilera. An. Acad. Bras. Cien. 28: 369-371. 1956.

HAYASHI, M.; NASCIMENTO, F.D.; KERKIS, A.; OLIVEIRA, V.; OLIVEIRA, E.B.; PEREIRA, A.; RADIS-BAPTISTA, G.; NADER, H.; YAMANE, T.; KERKIS, I.; TERSARIOL, I. Cytotoxic effects of crotamine are mediated through lysosomal membrane permeabilization. Toxicon. 52: 508-517. 2008.

HERNÁNDEZ, M.; SCANNONE, H.; FINOL, H.J.; PINEDA, M.E.; FERNÁNDEZ, I.; GIRÓN, M.E.; AGUILAR, I; RODRÍGUEZ-ACOSTA, A. The crotoxin activity of rattlesnake venom (Crotalus durissus cumanensis) on the ultrastructure of cardiac atrial muscle. Exp. Toxicol. Pathol. 59: 129-137. 2007.

JOLLIFFE, I.T.; CADIMA, J. Principal component analysis: a review and recent developments. Philos. Trans. A. Math. Phys. Eng. Sci. 374(2065): 2015-2020. 2016.

KASTURIRATNE, A.; WICKREMASINGHE, R.; DE SILVA, N.; GUNAWARDENA, K.; PATHMESWARAN, A.; PREMARATNA, R.; SAVIOLI, L.; LALLOO, D.; DE SILVA, J. The global burden of snakebite: A literature analysis and modelling base on regional estimates of envenoming and deaths. PLoS Med. 5: e218. 2008.

KERKIS, I.; HAYASHI, M.A.; PRIETO, D.A.; SILVA, A.R.; PEREIRA, A.; DE SÁ JR, P.L.; ZAHARENKO, A.J.; RÁDIS-BAPTISTA, G.; KERKIS, A.; YAMANE, T. State of the art in the studies on crotamine, a cell penetrating peptide from South American rattlesnake. Biomed. Res. Int. 2014: 675985. 2014.

KLOSS, R.; GROSS, M.; FRANCIS, I.; DOROKIN, M.; SHAPIRO, B. Incidentally discovered adrenal mases. Endocrine. Rev. 16: 460-484. 1995.

KONDRATSKYI, A.; KONDRATSKA, K.; SKRYMA, R.; PREVARSKAYA, N. Ion channels in the regulation of apoptosis. Biochim. Biophys. Acta. 1848: 2532-2546. 2015.

LLOYD, R.; DOUGLAS, B.; YOUNG, W. Adrenal Gland. Endocrine disease. Atlas of non-tumor pathology: Editorial King D: AFIP, Washington DC (USA). Pp. 209-212. 2002.

MONTGOMERY, D.C.; RUNGER, G.C. Probabilidades. Probabilidad y Estadística Aplicadas a la Ingeniería: McGraw-Hill, México, 895pp. 1996.

NASCIMENTO, J.M.; FRANCHI, G.C. JR.; NOWILL, A.E.; COLLARES-BUZATO, C.B.; HYSLOP, S. Cytoskeletal rearrangement and cell death induced by Bothrops alternatus snake venom in cultured Madin-Darby canine kidney cells. Biochem. Cell. Biol. 85(5): 591-605. 2007.

NICASTRO, G.; FRANZONI, L.; DE CHIARA, C.; MANCIN, A.; GIGLIO, J.; SPISNI, A. Solution structure of crotamine, a Na+ channel affecting toxin from Crotalus durissus terrificus venom. Eur. J. Biochem. 270: 1969-1979. 2003.

OGUIURA, N.; BONI-MITAKE, M.; RADIS-BAPTISTA, G. New view on crotamine, a small basic polypeptide myotoxin from South American rattlesnake venom. Toxicon. 46: 363–370. 2005.

PEIGNEUR, S.; ORTS, D.J.B.; DA SILVA, A.R.P.; OGUIURA, N.; BONI-MITAKE, M.; DE OLIVEIRA, E.B.; ZAHARENKO, A.J.; DE FREITAS, J.C.; TYTGAT, J. Crotamine pharmacology revisited: Novel insights based on the inhibition of KV channels. Mol. Pharmacol. 82: 90–96. 2012.

PONCE-SOTO, L.A.; MARTINS, D.; MARANGONI, S. Structural and Pharmacological Characterization of the Crotamine Isoforms III-4 (MYX4_CROCu) and III-7 (MYX7_CROCu) Isolated from the Crotalus durissus cumanensis Venom. Toxicon. 55: 1443-1452. 2010.

PONCE-SOTO, L.A.; MARTINS-DE-SOUZA, D.; NOVELLO, J.C.; MARANGONI, S. Structural and biological characterization of two crotamine isoforms IV-2 and IV-3 isolated from the Crotalus durissus cumanensis venom. Protein J. 26: 533-540. 2007.

PULIDO, M.; RODRIGUEZ-ACOSTA, A.; FINOL, H.J.; AGUILAR, I.; GIRÓN, M.E. Ultrastructural pathology in skeletal muscle of mice envenomed with Crotalus vegrandis venom. J. Submicrosc. Cytol. Pathol. 31: 555-561. 1999.

RADIS-BAPTISTA, G.; OGUIURA, N.; HAYASHI, M.A.F.; CAMARGO, M.E,; GREGO, K.F.; OLIVEIRA, E.B.; YAMANE, T. Nucleotide sequence of crotamine isoform precursors from a single South American rattlesnake (Crotalus durissus terrificus). Toxicon. 37: 973-984. 1999.

RENGIFO, C.; RODRÍGUEZ-ACOSTA, A. Capitulo III. Serpientes veneno y tratamiento médico en Venezuela: Fondo Editorial de la Facultad de Medicina, Universidad Central de Venezuela Caracas, Venezuela. 272 pp. 2019.

REYES-JUÁREZ, J.L.; ZARAIN-HERZBERG, Á. Función del retículo sarcoplásmico y su papel en las enfermedades cardíacas. Arch. Cardiol. Mex. 76: 18-32. 2006.

SÁNCHEZ, E.E.; GONZÁLEZ, R.; LUCENA, S.; GARCÍA, S.; FINOL, H.J.; SUNTRAVAT, M.; GIRÓN, M.E.; FERNÁNDEZ, I.; RODRÍGUEZ-ACOSTA, A. Crotamine-like from Southern Pacific rattlesnake (Crotalus oreganus helleri) Venom acts on human leukemia (K-562) cell lines and produces ultrastructural changes on mice adrenal gland. Ultrastruc. Pathol. 42: 116–123. 2018.

SIEBER, M.; BOSCH, B.; HANKE, W.; FERNANDES-DE LIMA, V.M. Membrane-modifying properties of crotamine, a small peptide-toxin from Crotalus durissus terrificus venom. Biochim. Biophys. Acta. 1840: 945-950. 2014.

SOKAL, R.R.; ROHLF, F.J. Biometry. The Principles and Practice of Statistics in Biological Research: Freeman WH and Company San Francisco (USA), 253 pp. 2012.

SOULSBY, E.J. The Sir Frederick Hobday Memorial Lecture. We don’t shoot horses anymore. Equine Vet. J. 20 (Supl. 4): 243-248. 1981.

STOSCHECK, C.M. Quantitation of protein. Methods. Enzymol. 182:50–68. 1990.

TISHENINA, R.S.; KALININ, A.P.; KOZLOVA, N.K. Bolezn’ Itsenko-Kushinga i perekisnoe okislenie lipidov [Itsenko-Cushing disease and lipid peroxidation]. Probl. Endokrinol. (Mosk). 34: 32-36. 1988.

TOYAMA, M.H.; CARNEIRO, E.M.; MARANGONI, S.; BARBOSA, R.L.; CORSO, G.; BOSCHERO, A.C. Biochemical characterization of two crotamine isoforms isolated by a single step RP-HPLC from Crotalus durissus terrificus (South American rattlesnake) venom and their action on insulin secretion by pancreatic islets. Biochim. Biophys. Acta. 1474: 56-60. 2000.

WANG, X.; KOMATSU, S. Isolation, purity assessment, and proteomic analysis of Endoplasmic Reticulum. Methods. Mol. Biol. 2139: 117-131. 2020.

WANG, Y.; HEKIMI, S. Mitochondrial dysfunction and longevity in animals: Untangling the knot. Sci. 350: 1204-1207. 2015.

BRUNI, V.; CARDINALI, M.L. VITULANO’, D. A Short Review on Minimum Description Length: An Application to Dimension Reduction in PCA. Entropy (Basel). 24(2): 269-285. 2022.

Published
2022-05-18
How to Cite
1.
García E, Finol† HJ, González R, Rodríguez-Acosta A. A new crotamine-like from the rattlesnake (Crotalus durissus cumanensis) venom causing damages: Qualitative and Quantitative Cytotoxic Studies on subcellular and neuromuscular structures. Rev. Cient. FCV-LUZ [Internet]. 2022May18 [cited 2024Nov.25];32:1-13. Available from: https://produccioncientificaluz.org/index.php/cientifica/article/view/38134
Section
Veterinary Medicine