title: Morphological and ultrastructural effects of microcystin-LR from Microcystis aeruginosa extract on a kidney cell line.
abstract: The aim of this study was to examine the toxic effects of a microcystin-LR (MCLR)-containing cyanobacteria extract on the subcellular organization of a kidney cell line (Vero-E6). Cells were exposed to different MCLR concentrations (1.3-150 microM) for 24, 48 and 72h and two cytotoxicity assays were performed. This information was combined with the analysis of lysosomal, mitochondrial and cytoskeleton integrity and with an ultrastructural study. Biochemical and microscopic data revealed a good agreement and demonstrated that cellular response to MCLR is dependent on the dose/exposure time. Cell viability decayed markedly after 24h of exposure to toxin concentrations greater than 30 microM. Furthermore, it was demonstrated that lysosome destabilization precedes mitochondria dysfunction. The ultrastructural analysis showed that mild toxin incubation conditions induce endoplasmic reticulum (ER) vacuolization and assembly of large autophagic vacuoles, suggesting that autophagy is an early cellular response to the toxin. After exposure to higher MCLR doses, the number of apoptotic cells increased, as identified by microscopic observations and confirmed with TUNEL assay. Additionally, drastic exposure conditions induced the increase of necrotic cells. These results suggest that the ER is the primary microcystin target in Vero cells and that autophagy, apoptosis and necrosis are induced in a dose- and time-dependent manner.
authors.affiliation: Department of Environmental Health, National Institute of Health Dr Ricardo Jorge, 1649-016 Lisbon, Portugal. Elsa.Alverca@insa.min-saude.pt
authors.forename: E
authors.initials: E
authors.lastname: Alverca
authors.forename: M
authors.initials: M
authors.lastname: Andrade
authors.forename: E
authors.initials: E
authors.lastname: Dias
authors.forename: F
authors.initials: F
authors.lastname: Sam Bento
authors.forename: M C C
authors.initials: MC
authors.lastname: Batoréu
authors.forename: P
authors.initials: P
authors.lastname: Jordan
authors.forename: M J
authors.initials: MJ
authors.lastname: Silva
authors.forename: P
authors.initials: P
authors.lastname: Pereira
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D002460:Cell Line; D002522:Chlorocebus aethiops; D020287:In Situ Nick-End Labeling; D007668:Kidney / Q000166:cytology / Q000187:drug effects* / Q000648:ultrastructure; D008387:Marine Toxins; D052998:Microcystins / Q000633:toxicity*; D046931:Microcystis / Q000737:chemistry*; D008856:Microscopy, Fluorescence; D014709:Vero Cells
chemical_list: D008387:Marine Toxins; D052998:Microcystins; C057862:cyanoginosin LR