Flow cytometric measurement of reactive oxygen species production in red blood cells from fish after exposure to heavy metals

Flow cytometric measurement of reactive oxygen species production in red blood cells from fish after exposure to heavy metals

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Title: Flow cytometric measurement of reactive oxygen species production in red blood cells from fish after exposure to heavy metals
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Article_Title: Flow cytometric measurement of reactive oxygen species production in red blood cells from fish after exposure to heavy metals
Authors: Aurelia Covaci, Ciprian-Valentin Mihali, Violeta Turcuş, Daniela Bratosin
Affiliation: Faculty of Natural Sciences, Engineering and Informatics, ″V. Goldis″ Western University of Arad, Romania
Institute of Life Sciences, ″Vasile Goldis″ Western University of Arad, Romania
National Institute for Biological Science Research & Development (INCDSB), Bucharest, Romania
Abstract: Organisms react to environmental pollutants by disturbance of living processes at subcellular levels resulting in cells death. To understand the mechanisms underlying the process of cell death by heavy metals action, we measured by flow cytometry the reactive oxygen species (ROS) generation by heavy metal (Al, Cd, Hg, Pb, Zn) in fish nucleated erythrocytes correlated with morphological changes by scattered light flow cytometry in the mode FSC/SSC. Optical and scanning electron microscopy were accompanied the research. The results we obtained show that heavy metals generates a significant amount of ROS that induce significant morphological changes characteristic for the phenomenon of cell death by apoptosis. The present results demonstrate that oxidative stress assessment and the use of fish nucleated erythrocytes “in vitro”, can be a good tool for ecotoxicological evaluation of the presence of pollutants, heavy metals in particular.
Keywords: nucleated erythrocytes, fish heavy metals, apoptosis, reactive oxygen species, flow cytometry, ESEM, optic macroscopy, ecotoxicology
References: Bratosin D., Estaquier J., Petit F., Tissier JP., Trandaburu I., Slomianny C., Huart JJ., Ameisen JC., Montreuil J., Molecular and cellular mechanism of erythrocyte cell death. An apoptotic phenomenon. Biochimiem, 81, p. 361, 1999.
Bratosin D., Estaquier J., Slomianny C., Tissier J-P, Quatannes B., Bulai T., Mitrofan L., Marinescu A., Trandaburu I., Ameisen J-C, Montreuil J., On the evolution of erythrocyte programmed cell death: apoptosis of Rana esculenta nucleated red blood cells involves cysteine proteinase activation and mitochondrion permeabilization, Biochimie, 86, pp. 183-193, 2004.
Bratosin D., Takacs L., Gheorghe A.-M., Sidoroff M., Tcacenko L., Ponepal C., Buruiana V., Hermenean A., Ardelean A., Boscaiu V., Montreuil J., Marinescu Al. G., Flow cytometric measurement of reactive oxygen species produced in aluminum mediated-apoptosis of Rana nucleated erythrocytes, Romanian Biological Sciences. vol V, 1-4, pp. 29-36, 2007.
Di Giulio RT., Meyer J.N., Reactive oxygen species and oxidative stress, In: Di Giulio RT, Hinton DE (eds.): The Toxicology of Fishes, CRC Press, Taylor and Francis Group, 273–324, 2008.
Jezierska B., Witeska M., Summary of metal-induced disturbances in fish organism. In: Metal Toxicity to Fish, Wydawnictvo Akademii Podlaskej, Siedlce, 214–243, 2001.
Kamunde C., Clayton C., Wood CM., Waterborne vs. dietary copper uptake in rainbow trout and the effects of previous waterborne copper exposure. American Journal of Physiology – Regulatory, Integrative and Comparative Physiology, 283, R69–R78, 2002.
Livingstone DR., Contaminant-stimulated reactive oxygen species production and oxidative damage in aquatic organisms, Mar. Pollut. Bull., 42, 656-666, 2001.
Livingstone DR., Oxidative stress in aquatic organism in relation to pollution and agriculture, Revue de Medecine Veterinaire 154, 427–430, 2003.
Luoma SN., Rainbow PS., Sources and cycles of trace metals, In: Metal Contamination in Aquatic Environments: Science and Lateral Management. Cambridge University Press, Cambridge, 47–66, 2008a.
Nishida Y., The chemical process of oxidative stress by copper (II) and iron (III) ions in several neurodegenerative disorders, Monatshefte fur Chemie, 142, 375–384, 2011.
Slaninova A., Smutna M., Modra H., Svobodova Z., A review: Oxidative stress in fish induced by pesticides, Neuroendocrinology Letters, 30, 2–12, 2009.
Tao S., Wen Y., Long A., Dawson R., Cao J., Xu F., Simulation of acid-base condition and copper speciation in fish gill microenvironment, Computers and Chemistry, 25, 215–222, 2001.
Read_full_article: pdf/23-2013/23-3-2013/SU23-3-2013-Covaci1.pdf
Correspondence: Daniela Bratosin, National Institute for Biological Science Research & Development, Bucharest, Romania; Splaiul Independentei no. 296, district 6, Bucharest, Romania, Tel/Fax +40-(021)-2200881, email:bratosind@yahoo.com

Read full article
Article Title: Flow cytometric measurement of reactive oxygen species production in red blood cells from fish after exposure to heavy metals
Authors: Aurelia Covaci, Ciprian-Valentin Mihali, Violeta Turcuş, Daniela Bratosin
Affiliation: Faculty of Natural Sciences, Engineering and Informatics, ″V. Goldis″ Western University of Arad, Romania
Institute of Life Sciences, ″Vasile Goldis″ Western University of Arad, Romania
National Institute for Biological Science Research & Development (INCDSB), Bucharest, Romania
Abstract: Organisms react to environmental pollutants by disturbance of living processes at subcellular levels resulting in cells death. To understand the mechanisms underlying the process of cell death by heavy metals action, we measured by flow cytometry the reactive oxygen species (ROS) generation by heavy metal (Al, Cd, Hg, Pb, Zn) in fish nucleated erythrocytes correlated with morphological changes by scattered light flow cytometry in the mode FSC/SSC. Optical and scanning electron microscopy were accompanied the research. The results we obtained show that heavy metals generates a significant amount of ROS that induce significant morphological changes characteristic for the phenomenon of cell death by apoptosis. The present results demonstrate that oxidative stress assessment and the use of fish nucleated erythrocytes “in vitro”, can be a good tool for ecotoxicological evaluation of the presence of pollutants, heavy metals in particular.
Keywords: nucleated erythrocytes, fish heavy metals, apoptosis, reactive oxygen species, flow cytometry, ESEM, optic macroscopy, ecotoxicology
References: Bratosin D., Estaquier J., Petit F., Tissier JP., Trandaburu I., Slomianny C., Huart JJ., Ameisen JC., Montreuil J., Molecular and cellular mechanism of erythrocyte cell death. An apoptotic phenomenon. Biochimiem, 81, p. 361, 1999.
Bratosin D., Estaquier J., Slomianny C., Tissier J-P, Quatannes B., Bulai T., Mitrofan L., Marinescu A., Trandaburu I., Ameisen J-C, Montreuil J., On the evolution of erythrocyte programmed cell death: apoptosis of Rana esculenta nucleated red blood cells involves cysteine proteinase activation and mitochondrion permeabilization, Biochimie, 86, pp. 183-193, 2004.
Bratosin D., Takacs L., Gheorghe A.-M., Sidoroff M., Tcacenko L., Ponepal C., Buruiana V., Hermenean A., Ardelean A., Boscaiu V., Montreuil J., Marinescu Al. G., Flow cytometric measurement of reactive oxygen species produced in aluminum mediated-apoptosis of Rana nucleated erythrocytes, Romanian Biological Sciences. vol V, 1-4, pp. 29-36, 2007.
Di Giulio RT., Meyer J.N., Reactive oxygen species and oxidative stress, In: Di Giulio RT, Hinton DE (eds.): The Toxicology of Fishes, CRC Press, Taylor and Francis Group, 273–324, 2008.
Jezierska B., Witeska M., Summary of metal-induced disturbances in fish organism. In: Metal Toxicity to Fish, Wydawnictvo Akademii Podlaskej, Siedlce, 214–243, 2001.
Kamunde C., Clayton C., Wood CM., Waterborne vs. dietary copper uptake in rainbow trout and the effects of previous waterborne copper exposure. American Journal of Physiology – Regulatory, Integrative and Comparative Physiology, 283, R69–R78, 2002.
Livingstone DR., Contaminant-stimulated reactive oxygen species production and oxidative damage in aquatic organisms, Mar. Pollut. Bull., 42, 656-666, 2001.
Livingstone DR., Oxidative stress in aquatic organism in relation to pollution and agriculture, Revue de Medecine Veterinaire 154, 427–430, 2003.
Luoma SN., Rainbow PS., Sources and cycles of trace metals, In: Metal Contamination in Aquatic Environments: Science and Lateral Management. Cambridge University Press, Cambridge, 47–66, 2008a.
Nishida Y., The chemical process of oxidative stress by copper (II) and iron (III) ions in several neurodegenerative disorders, Monatshefte fur Chemie, 142, 375–384, 2011.
Slaninova A., Smutna M., Modra H., Svobodova Z., A review: Oxidative stress in fish induced by pesticides, Neuroendocrinology Letters, 30, 2–12, 2009.
Tao S., Wen Y., Long A., Dawson R., Cao J., Xu F., Simulation of acid-base condition and copper speciation in fish gill microenvironment, Computers and Chemistry, 25, 215–222, 2001.
*Correspondence: Daniela Bratosin, National Institute for Biological Science Research & Development, Bucharest, Romania; Splaiul Independentei no. 296, district 6, Bucharest, Romania, Tel/Fax +40-(021)-2200881, email:bratosind@yahoo.com