Abstract
The mycotoxin ochratoxin A (OTA) is a naturally occuring contaminant of food. The genotoxic status of OTA is still controversial because contradictory results were obtained in various microbial and mammalian gene mutation assays. In this study, OTA was investigated to examine its potency to induce micronuclei (MN) in SHE cells. The SHE-micronucleus assay revealed that OTA induces MN in a dose- and time-dependent manner. The results of kinetochore analysis revealed that mainly clastogenic events are involved in OTA genotoxicity. Induction of mitotic disturbances can be closely related to changes of the intracellular calcium concentration ([Ca 2+] i). The investigated time course of OTA-induced [Ca 2+] i changes revealed that the obtained signal is a short spike signal resembling physiological responses. In the absence of extracellular calcium, a long-lasting signal indicates possible damage to intracellular calcium stores or channels. Our data show that the OTA-induced [Ca 2+] i rise is caused by Ca 2+-release from intracellular stores as well as Ca 2+ influx from extracellular area. Finally, the influence of the changed intracellular calcium level on the actin cytoskeleton was investigated. Visualization of the actin filaments revealed time- and concentration-dependent effects. Cell shrinkage and depolymerized filaments were observed. We conclude that OTA disrupts actin filaments by a direct irreversible binding to actin.
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