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Generation of reactive oxygen species by leukocytes of Prochilodus lineatus

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Abstract

Prochilodus lineatus (curimbatá), from the Procholodontidae family, is a Brazilian freshwater fish, which is important commercially, nutritionally and ecologically. It is encountered in the Rio da Prata Bay in Southern South America. Studies on the immune system of this fish are scarce, but the physiological mechanisms of the species are analogous to those of other vertebrates. Thus, this work discusses the present study, which correlates P. lineatus leukocytes and the generation of reactive oxygen species after modulatory stimuli. Leukocytes were characterized by light and electron transmission microscopy and investigated by the generation of H2O2 and O2 −, using phenol red, flow-cytometry and electron transmission histochemistry. The study determined that monocytes and neutrophils are the main cells responsible for generating O2 after stimulation with phorbol myristate acetate. Superoxide dismutase successfully inhibited the generation of reactive oxygen species in neutrophils and monocytes, but stimulated generation when in association with phorbol myristate acetate. Fish leukocyte samples from P. lineatus showed cross-reactivity with antibodies directed against human NADPH-oxidase antibody subunits (p47phox and p67phox). Thus, catalase enhanced the presence of p47phox. Neutrophil mitochondria were shown to be generators of H2O2 (charged by cerium precipitate), being enlarged and changing their format. The present study contributes to a better understanding of the respiratory burst pathways in this species and suggests mitochondria as the organelle responsible for generation of reactive oxygen species.

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Acknowledgments

The authors are indebted to Prof. Maristela M.C. Camargo and technician Andrea Glatt for their help with the FACS Vantage®, Sidney Veríssimo Filho for his help with Western blot techniques, Edson de Oliveira for his help with photography, Cláudia Fernanda Pantoja da Silva for her help with reference review, and FAPESP grant (2001/08205-3), CNPq and CAPES for financial support.

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Authors and Affiliations

  1. Brazilian Agricultural Research Corporation, Embrapa, Trav. Dr. Enéas Pinheiro S/N, Caixa Postal 48, Belém, PA, 66095-130, Brazil

    Marcos Tucunduva de Faria

  2. Department of Physiology and Biophysics, Biomedical Sciences Institute, University of São Paulo, São Paulo, Brazil

    Maria Fernanda Cury-Boaventura

  3. Department of Pharmacology, Biomedical Sciences Institute, University of São Paulo, São Paulo, Brazil

    Lucia Rossetti Lopes

  4. Department of Cellular and Developmental Biology, Biomedical Sciences Institute and CEBIMar, Marine Biology Center, University of São Paulo, São Paulo, Brazil

    José Roberto Machado Cunha da Silva

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  1. Marcos Tucunduva de Faria
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  2. Maria Fernanda Cury-Boaventura
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  3. Lucia Rossetti Lopes
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  4. José Roberto Machado Cunha da Silva
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Correspondence to Marcos Tucunduva de Faria.

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de Faria, M.T., Cury-Boaventura, M.F., Lopes, L.R. et al. Generation of reactive oxygen species by leukocytes of Prochilodus lineatus . Fish Physiol Biochem 40, 445–455 (2014). https://doi.org/10.1007/s10695-013-9856-9

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