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Published on: September 1, 2015
Morphological and ultrastructural effects of microcystin-LR from Microcystis aeruginosa extract on a kidney cell line
1Department of Environmental Health, National Institute of Health Dr Ricardo Jorge, 1649-016 Lisbon, Portugal. Elsa.Alverca@insa.min-saude.pt
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.
Insights
Microcystin-LR (MCLR) from cyanobacteria extract damages kidney cells by disrupting cellular structures. Autophagy, apoptosis, and necrosis occur in a dose- and time-dependent manner, with the endoplasmic reticulum being the primary target.
Area of Science:
- Toxicology
- Cell Biology
- Environmental Health
Background:
- Cyanobacteria blooms produce toxins like microcystin-LR (MCLR).
- MCLR is a potent hepatotoxin, but its effects on kidney cells are less understood.
- Kidney cell lines provide a model to study cellular responses to toxins.
Purpose of the Study:
- To investigate the toxic effects of MCLR on the subcellular organization of Vero-E6 kidney cells.
- To determine the dose- and time-dependent cellular responses to MCLR exposure.
- To identify the primary cellular targets and pathways involved in MCLR toxicity.
Main Methods:
- Exposure of Vero-E6 cells to varying MCLR concentrations (1.3-150 microM) for 24, 48, and 72 hours.
- Cytotoxicity assays, lysosomal, mitochondrial, and cytoskeleton integrity analyses.
- Ultrastructural studies, including TUNEL assay for apoptosis detection.
Main Results:
- Cell viability decreased significantly at MCLR concentrations above 30 microM after 24 hours.
- Lysosomal destabilization was observed before mitochondrial dysfunction.
- Endoplasmic reticulum vacuolization and autophagic vacuole formation indicated early autophagy induction.
- Apoptosis and necrosis increased with higher MCLR doses and longer exposure times.
Conclusions:
- The endoplasmic reticulum is the primary target of MCLR in Vero cells.
- Autophagy, apoptosis, and necrosis are key cellular responses to MCLR toxicity.
- Cellular damage is intricately linked to MCLR dose and exposure duration.
