PURPOSE: The goal of this study was to investigate the modifications of aquaporin (AQP) expression in ARPE-19 cells in response to fenretinide-induced transdifferentiation into neuronal-like cells METHODS: ARPE-19 cells were treated daily for 7 days with 3 μm fenretinide or dimethyl sulphoxide as control. mRNA and protein expression were evaluated by real-time quantitative PCR, Western blot analysis and immunofluorescence. RESULTS: Control ARPE-19 cells expressed AQP1, AQP4, AQP6 and AQP11 at the mRNA level, but only AQP4, AQP6 and AQP11 at the protein level. Fenretinide induced the transdifferentiation of ARPE-19 cells into neuronal-like cells. Indeed, fenretinide induced morphological changes similar to neurons characterized by elongated cell body and the formation of neurite branching. Moreover, ARPE-19 cells transdifferentiated to neuron-like cells were characterized by significant decrease in retinal pigmented epithelium markers, for example cytokeratin 8 and cellular retinaldehyde-binding protein, as well as an increase in neuronal markers such as synaptophysin and calretinin. AQP4 expression, at both mRNA and protein levels, and AQP6 expression, only at protein level, were significantly decreased in ARPE-19 cells transdifferentiated into neuronal-like cells. CONCLUSIONS: The expression of AQP4 and AQP6 is downregulated during fenretinide-induced transdifferentiation.
PURPOSE: The goal of this study was to investigate the modifications of aquaporin (AQP) expression in ARPE-19 cells in response to fenretinide-induced transdifferentiation into neuronal-like cells METHODS: ARPE-19 cells were treated daily for 7 days with 3 μm fenretinide or dimethyl sulphoxide as control. mRNA and protein expression were evaluated by real-time quantitative PCR, Western blot analysis and immunofluorescence. RESULTS: Control ARPE-19 cells expressed AQP1, AQP4, AQP6 and AQP11 at the mRNA level, but only AQP4, AQP6 and AQP11 at the protein level. Fenretinide induced the transdifferentiation of ARPE-19 cells into neuronal-like cells. Indeed, fenretinide induced morphological changes similar to neurons characterized by elongated cell body and the formation of neurite branching. Moreover, ARPE-19 cells transdifferentiated to neuron-like cells were characterized by significant decrease in retinal pigmented epithelium markers, for example cytokeratin 8 and cellular retinaldehyde-binding protein, as well as an increase in neuronal markers such as synaptophysin and calretinin. AQP4 expression, at both mRNA and protein levels, and AQP6 expression, only at protein level, were significantly decreased in ARPE-19 cells transdifferentiated into neuronal-like cells. CONCLUSIONS: The expression of AQP4 and AQP6 is downregulated during fenretinide-induced transdifferentiation.
Authors: Cornelia A Deeg; Barbara Amann; Konstantin Lutz; Sieglinde Hirmer; Karina Lutterberg; Elisabeth Kremmer; Stefanie M Hauck Journal: J Neuroinflammation Date: 2016-04-23 Impact factor: 8.322