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Microcystin-LR affects cytoskeleton and morphology of rabbit primary whole embryo cultured cells in vitro
Authors:R Frangez  M C Zuzek  J Mrkun  D Suput  B Sedmak  M Kosec
Institution:

a Veterinary Faculty, Institute of Physiology, Pharmacology and Toxicology, University of Ljubljana, Slovenia

b National Institute of Biology, University of Ljubljana, Slovenia

c Medical Faculty, Institute of Pathophysiology, University of Ljubljana, Slovenia

Abstract:Microcystin-LR is the most frequently studied cyclic heptapeptide produced by different genera of cyanobacteria and is hepatotoxic to livestock and human populations. The adverse effects of microcystin-LR on morphology and cytoskeletal elements in different stages of early embryonal development have been studied in vitro. Embryos and whole embryo cultures have been exposed to microcystin-LR (10–100 μM). Actin filaments were visualized by fluorescence staining and the microtubular network labelled by immunostaining. Growth, development and cytoskeleton organization of the embryos embedded in zona pellucida are not affected by microcystin-LR in concentrations up to 100 μM, while whole embryo cell cultures are affected by the presence of microcystin-LR in the culture medium. High microcystin-LR concentrations (100 μM) cause cells to be detached and destroyed, while lower concentrations (10–20 μM) profoundly affect actin and microtubule organization. These effects are confirmed also by the presence of transformed microcystin-LR in all the media at the lowest concentrations. It seems that the changes to the cells are far more serious than that expressed in cell morphology. From our experiments we conclude that the presence of zona pellucida is an effective way of embryo protection against xenobiotics like microcystin-LR.
Keywords:Cyanobacteria  Microcystin-LR  Rabbit embryo  Actin filaments  Microtubules  Cytoskeleton  Laser scanning microscope
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