title: 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.
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.
authors.affiliation: Université Reims Champagne-Ardenne, EA Unité Interactions Animal-Environnement, Moulin de Housse, BP 1039, 51687 Reims, France. anne.bado-nilles@ineris.fr
authors.forename: Anne
authors.initials: A
authors.lastname: Bado-Nilles
authors.forename: Stéphane
authors.initials: S
authors.lastname: Betoulle
authors.forename: Alain
authors.initials: A
authors.lastname: Geffard
authors.forename: Jean-Marc
authors.initials: JM
authors.lastname: Porcher
authors.forename: Béatrice
authors.initials: B
authors.lastname: Gagnaire
authors.forename: Wilfried
authors.initials: W
authors.lastname: Sanchez
mesh_terms: D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D003300:Copper / Q000633:toxicity; D004726:Endosulfan / Q000633:toxicity; D004784:Environmental Monitoring / Q000379:methods*; D005434:Flow Cytometry / Q000379:methods*; D005602:France; D007306:Insecticides / Q000633:toxicity; D007425:Intracellular Membranes / Q000187:drug effects*; D007962:Leukocytes / Q000187:drug effects*; D008247:Lysosomes / Q000187:drug effects*; D023701:Smegmamorpha / Q000276:immunology / Q000502:physiology*; D013154:Spleen / Q000187:drug effects; D014874:Water Pollutants, Chemical / Q000633:toxicity*
chemical_list: D015415:Biomarkers; D007306:Insecticides; D014874:Water Pollutants, Chemical; D003300:Copper; D004726:Endosulfan