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      Toxicological effects of bioactive peptide fractions obtained from Bothrops jararaca snake venom on the structure and function of mouse seminiferous epithelium

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          Abstract

          Background:

          Pathogenesis of Bothrops envenomations is complex and despite numerous studies on the effects of this snake venom on various biological systems, relatively little is known about such effects on the male reproductive system. In the present study, the toxicological outcomes of the low molecular weight fraction (LMWF) of B. jararaca snake venom - containing a range of bioactive peptides - were investigated on the dynamics and structure of the seminiferous epithelium and 15P-1 Sertoli cells viability.

          Methods:

          LMWF (5 µg/dose per testis) venom was administered in male Swiss mice by intratesticular (i.t.) injection. Seven days after this procedure, the testes were collected for morphological and morphometric evaluation, distribution of claudin-1 in the seminiferous epithelium by immunohistochemical analyses of testes, and the nitric oxide (NO) levels were evaluated in the total extract of the testis protein. In addition, the toxicological effects of LMWF and crude venom (CV) were analyzed on the 15P-1 Sertoli cell culture.

          Results:

          LMWF induced changes in the structure and function of the seminiferous epithelium without altering claudin-1 distribution. LMWF effects were characterized especially by lost cells in the adluminal compartment of epithelium (spermatocytes in pachytene, preleptotene spermatocytes, zygotene spermatocytes, and round spermatid) and different stages of the seminiferous epithelium cycle. LMWF also increased the NO levels in the total extract of the testis protein and was not cytotoxic in concentrations and time tested in the present study. However, CV showed cytotoxicity at 10 μg/mL from 6 to 48 h of treatment.

          Conclusions:

          The major finding of the present study was that the LMWF inhibited spermatozoa production; principally in the spermiogenesis stage without altering claudin-1 distribution in the basal compartment. Moreover, NO increased by LMWF induce open of complexes junctions and release the germ cells of the adluminal compartment to the seminiferous tubule.

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          MTT assay for cell viability: Intracellular localization of the formazan product is in lipid droplets.

          Juan Stockert, Alfonso Blázquez-Castro, Magdalena Cañete … (2012)
          Although MTT is widely used to assess cytotoxicity and cell viability, the precise localization of its reduced formazan product is still unclear. In the present study the localization of MTT formazan was studied by direct microscopic observation of living HeLa cells and by colocalization analysis with organelle-selective fluorescent probes. MTT formazan granules did not colocalize with mitochondria as revealed by rhodamine 123 labeling or autofluorescence. Likewise, no colocalization was observed between MTT formazan granules and lysosomes labeled by neutral red. Taking into account the lipophilic character and lipid solubility of MTT formazan, an evaluation of the MTT reaction was performed after treatment of cells with sunflower oil emulsions to induce a massive occurrence of lipid droplets. Under this condition, lipid droplets revealed a large amount of MTT formazan deposits. Kinetic studies on the viability of MTT-treated cells showed no harmful effects at short times. Quantitative structure-activity relations (QSAR) models were used to predict and explain the localization of both the MTT tetrazolium salt and its formazan product. These predictions were in agreement with experimental observations on the accumulation of MTT formazan product in lipid droplets. Copyright © 2012 Elsevier GmbH. All rights reserved.
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            Spermatogenesis

            D.M. de Kretser, K.L Loveland, A Meinhardt … (1998)
            Article 0 comments Cited 104 times – based on 0 reviews     Review now
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              Epidemiological, clinical and therapeutic aspects of Bothrops asper bites.

              Rafael Otero-Patiño (2009)
              Bothrops asper inflicts the majority of snakebites in Central America and in the northern regions of South America, mostly affecting young agricultural workers in rural settings. This species is capable of provoking severe envenomings associated with local and systemic manifestations. The main clinical features are: local edema, ecchymoses, blisters, dermonecrosis, myonecrosis, defibrinogenation, thrombocytopenia, systemic bleeding, hypotension and renal alterations. In addition, soft-tissue infection, acute renal failure, compartmental syndrome, central nervous system hemorrhage and, in pregnant women, abortion, fetal wastage and abruptio placentae have been described as complications. Intravenous administration of antivenom constitutes the mainstay in the therapy. Antivenoms composed of either whole IgG or F(ab')(2) fragments, manufactured in Brazil, Colombia, Costa Rica and Mexico, have been tested in controlled clinical trials, and rational protocols for antivenom administration have been developed. In addition to antivenom therapy, a number of ancillary interventions are recommended in the treatment of B. asper bites.
              Article 0 comments Cited 85 times – based on 0 reviews     Review now
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                Author and article information

                Journal
                Journal ID (nlm-ta): J Venom Anim Toxins Incl Trop Dis
                Journal ID (iso-abbrev): J Venom Anim Toxins Incl Trop Dis
                Journal ID (publisher-id): jvatitd
                Title: The Journal of Venomous Animals and Toxins Including Tropical Diseases
                Publisher: Centro de Estudos de Venenos e Animais Peçonhentos
                ISSN (Electronic): 1678-9199
                Publication date (Electronic, pub): 22 June 2020
                Publication date (Electronic, collection): 2020
                Volume: 26
                Electronic Location Identifier: e20200007
                Affiliations
                [1 ]Natural and Humanities Sciences Center, Experimental Morphophysiology Laboratory, Federal University of ABC (UFABC), São Bernardo do Campo, SP, Brazil.
                [2 ]Department of Biophysics, Federal University of São Paulo (Unifesp), São Paulo, SP, Brazil.
                [3 ]Center for Health and Biological Sciences, Minas Gerais State University (UEMG), Passos, MG, Brazil.
                Author notes
                [* ]Correspondence: carlos.asilva@ufabc.edu.br

                Competing interests: The authors declare that they have no competing interests.

                Authors’ contributions: CAS established the conditions for the study, was responsible for drafting the manuscripts and reading the editorial corrections. JMG, CSF, and RSM carried out most experiments assisted by other researchers. SMQ contributed to the design of experiments and figures. All authors read and approved the final manuscript.

                Author information
                http://orcid.org/0000-0003-4519-8930
                Article
                Other ID: 00318
                DOI: 10.1590/1678-9199-JVATITD-2020-0007
                PMC ID: 7315626
                SO-VID: 8aa4b36b-4a25-40a1-9da8-f9261314a72f
                License:

                © The Author(s). 2020 Open Access This article is distributed under the terms of the Creative Commons Attribution 4.0 International License ( https://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver ( https://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

                History
                Date received : 14 January 2020
                Date accepted : 19 May 2020
                Page count
                Figures: 6, Tables: 0, Equations: 0, References: 47
                Categories
                Subject: Research

                Keywords: bothrops jararaca,snake venom,bradykinin-potentiating peptides,testis,seminiferous epithelium,15p-1 sertoli cells,spermatogenesis
                Data availability:
                Keywords: bothrops jararaca, snake venom, bradykinin-potentiating peptides, testis, seminiferous epithelium, 15p-1 sertoli cells, spermatogenesis

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