Flow cytometry detection of lysosomal presence and lysosomal membrane integrity in the three-spined stickleback (Gasterosteus aculeatus L.) immune cells: applications in environmental aquatic immunotoxicology

Anne Bado-Nilles 1 Stéphane Betoulle 2 Alain Geffard 2 Jean-Marc Porcher 1 Béatrice Gagnaire 3 Wilfried Sanchez 1
1 INERIS - Institut National de l'Environnement Industriel et des Risques
2 IAE - Unité de Recherche Interactions Animal-Environnement - EA 4689
3 Institut de Radioprotection et de Sûreté Nucléaire
Abstract : The neutral red retention assay has been proposed to determine the lysosomal membrane stability in immune cells. Nevertheless, this assay implies many examinations under a microscope at short time intervals and therefore the analysis of few samples. The present study proposes two more rapid, efficient, and sensitive sample analyses using flow cytometry method. Lysosomal presence and lysosomal membrane integrity (LMI) were evaluated on the three-spined stickleback, Gasterosteus aculeatus (L.), a well-described model fish species for aquatic ecotoxicology studies. After development of the two biomarkers, they were validated by ex vivo contamination with endosulfan and copper and by in situ sampling. These immunomarkers were clearly modulated by pollutants and their variations seemed to be correlated with leucocyte mortality. Thus, from a practical point of view, lysosomal presence and LMI may provide novel and efficient means of evaluating immune capacities and indicating the toxic effects of environmental pollution.
Keywords : FLOW CYTOMETRY LYSOSOMAL MEMBRANE INTEGRITY LYSOSOMAL PRESENCE POLLUTANT SPLEEN LEUCOCYTES THREE-SPINED STICKLEBACK (GASTEROSTEUS ACULEATUS L.)
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Submitted on : Friday, March 21, 2014 - 2:31:55 PM
Last modification on : Wednesday, March 27, 2019 - 9:16:03 AM

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Anne Bado-Nilles, Stéphane Betoulle, Alain Geffard, Jean-Marc Porcher, Béatrice Gagnaire, et al.. Flow cytometry detection of lysosomal presence and lysosomal membrane integrity in the three-spined stickleback (Gasterosteus aculeatus L.) immune cells: applications in environmental aquatic immunotoxicology. Environmental Science and Pollution Research, Springer Verlag, 2013, 20 (5), pp.2692-2704. ⟨10.1007/s11356-012-1410-2⟩. ⟨ineris-00963450⟩

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